Micro-computed tomography: Introducing new dimensions to taxonomy
Identifieur interne : 000442 ( Pmc/Corpus ); précédent : 000441; suivant : 000443Micro-computed tomography: Introducing new dimensions to taxonomy
Auteurs : Sarah Faulwetter ; Aikaterini Vasileiadou ; Michail Kouratoras ; Christos ArvanitidisSource :
- ZooKeys [ 1313-2989 ] ; 2013.
Abstract
Continuous improvements in the resolution of three-dimensional imaging have led to an increased application of these techniques in conventional taxonomic research in recent years. Coupled with an ever increasing research effort in cybertaxonomy, three-dimensional imaging could give a boost to the development of virtual specimen collections, allowing rapid and simultaneous access to accurate virtual representations of type material. This paper explores the potential of micro-computed tomography (X-ray micro-tomography), a non-destructive three-dimensional imaging technique based on mapping X-ray attenuation in the scanned object, for supporting research in systematics and taxonomy. The subsequent use of these data as virtual type material, so-called “cybertypes”, and the creation of virtual collections lie at the core of this potential. Sample preparation, image acquisition, data processing and presentation of results are demonstrated using polychaetes (bristle worms), a representative taxon of macro-invertebrates, as a study object. Effects of the technique on the morphological, anatomical and molecular identity of the specimens are investigated. The paper evaluates the results and discusses the potential and the limitations of the technique for creating cybertypes. It also discusses the challenges that the community might face to establish virtual collections. Potential future applications of three-dimensional information in taxonomic research are outlined, including an outlook to new ways of producing, disseminating and publishing taxonomic information.
Url:
DOI: 10.3897/zookeys.263.4261
PubMed: 23653515
PubMed Central: 3591762
Links to Exploration step
PMC:3591762Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Micro-computed tomography: Introducing new dimensions to taxonomy</title><author><name sortKey="Faulwetter, Sarah" sort="Faulwetter, Sarah" uniqKey="Faulwetter S" first="Sarah" last="Faulwetter">Sarah Faulwetter</name><affiliation><nlm:aff id="A1">Department of Zoology-Marine Biology, Faculty of Biology, National and Kapodestrian University of Athens, Panepistimiopolis, 15784, Athens, Greece,</nlm:aff></affiliation><affiliation><nlm:aff id="A2">Institute for Marine Biology, Biotechnology and Aquaculture, Hellenic Centre for Marine Research, 71003 Heraklion, Crete, Greece</nlm:aff></affiliation></author><author><name sortKey="Vasileiadou, Aikaterini" sort="Vasileiadou, Aikaterini" uniqKey="Vasileiadou A" first="Aikaterini" last="Vasileiadou">Aikaterini Vasileiadou</name><affiliation><nlm:aff id="A2">Institute for Marine Biology, Biotechnology and Aquaculture, Hellenic Centre for Marine Research, 71003 Heraklion, Crete, Greece</nlm:aff></affiliation><affiliation><nlm:aff id="A3">Department of Biology, University of Patras, 26504, Rio, Patras, Greece</nlm:aff></affiliation></author><author><name sortKey="Kouratoras, Michail" sort="Kouratoras, Michail" uniqKey="Kouratoras M" first="Michail" last="Kouratoras">Michail Kouratoras</name><affiliation><nlm:aff id="A4">Hellenic Centre for Marine Research, 71003 Heraklion, Crete, Greece</nlm:aff></affiliation></author><author><name sortKey="Arvanitidis, Christos" sort="Arvanitidis, Christos" uniqKey="Arvanitidis " first=" Christos" last="Arvanitidis"> Christos Arvanitidis</name><affiliation><nlm:aff id="A2">Institute for Marine Biology, Biotechnology and Aquaculture, Hellenic Centre for Marine Research, 71003 Heraklion, Crete, Greece</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">23653515</idno><idno type="pmc">3591762</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3591762</idno><idno type="RBID">PMC:3591762</idno><idno type="doi">10.3897/zookeys.263.4261</idno><date when="2013">2013</date><idno type="wicri:Area/Pmc/Corpus">000442</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Micro-computed tomography: Introducing new dimensions to taxonomy</title><author><name sortKey="Faulwetter, Sarah" sort="Faulwetter, Sarah" uniqKey="Faulwetter S" first="Sarah" last="Faulwetter">Sarah Faulwetter</name><affiliation><nlm:aff id="A1">Department of Zoology-Marine Biology, Faculty of Biology, National and Kapodestrian University of Athens, Panepistimiopolis, 15784, Athens, Greece,</nlm:aff></affiliation><affiliation><nlm:aff id="A2">Institute for Marine Biology, Biotechnology and Aquaculture, Hellenic Centre for Marine Research, 71003 Heraklion, Crete, Greece</nlm:aff></affiliation></author><author><name sortKey="Vasileiadou, Aikaterini" sort="Vasileiadou, Aikaterini" uniqKey="Vasileiadou A" first="Aikaterini" last="Vasileiadou">Aikaterini Vasileiadou</name><affiliation><nlm:aff id="A2">Institute for Marine Biology, Biotechnology and Aquaculture, Hellenic Centre for Marine Research, 71003 Heraklion, Crete, Greece</nlm:aff></affiliation><affiliation><nlm:aff id="A3">Department of Biology, University of Patras, 26504, Rio, Patras, Greece</nlm:aff></affiliation></author><author><name sortKey="Kouratoras, Michail" sort="Kouratoras, Michail" uniqKey="Kouratoras M" first="Michail" last="Kouratoras">Michail Kouratoras</name><affiliation><nlm:aff id="A4">Hellenic Centre for Marine Research, 71003 Heraklion, Crete, Greece</nlm:aff></affiliation></author><author><name sortKey="Arvanitidis, Christos" sort="Arvanitidis, Christos" uniqKey="Arvanitidis " first=" Christos" last="Arvanitidis"> Christos Arvanitidis</name><affiliation><nlm:aff id="A2">Institute for Marine Biology, Biotechnology and Aquaculture, Hellenic Centre for Marine Research, 71003 Heraklion, Crete, Greece</nlm:aff></affiliation></author></analytic><series><title level="j">ZooKeys</title><idno type="ISSN">1313-2989</idno><idno type="eISSN">1313-2970</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><label>Abstract</label><p>Continuous improvements in the resolution of three-dimensional imaging have led to an increased application of these techniques in conventional taxonomic research in recent years. Coupled with an ever increasing research effort in cybertaxonomy, three-dimensional imaging could give a boost to the development of virtual specimen collections, allowing rapid and simultaneous access to accurate virtual representations of type material. This paper explores the potential of micro-computed tomography (X-ray micro-tomography), a non-destructive three-dimensional imaging technique based on mapping X-ray attenuation in the scanned object, for supporting research in systematics and taxonomy. The subsequent use of these data as virtual type material, so-called “cybertypes”, and the creation of virtual collections lie at the core of this potential. Sample preparation, image acquisition, data processing and presentation of results are demonstrated using polychaetes (bristle worms), a representative taxon of macro-invertebrates, as a study object. Effects of the technique on the morphological, anatomical and molecular identity of the specimens are investigated. The paper evaluates the results and discusses the potential and the limitations of the technique for creating cybertypes. It also discusses the challenges that the community might face to establish virtual collections. Potential future applications of three-dimensional information in taxonomic research are outlined, including an outlook to new ways of producing, disseminating and publishing taxonomic information.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Abel, Rl" uniqKey="Abel R">RL Abel</name></author><author><name sortKey="Laurini, Cr" uniqKey="Laurini C">CR Laurini</name></author><author><name sortKey="Richter, M" uniqKey="Richter M">M Richter</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Alba Tercedor, J" uniqKey="Alba Tercedor J">J Alba-Tercedor</name></author><author><name sortKey="Sanchez Tocino, L" uniqKey="Sanchez Tocino L">L Sánchez-Tocino</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Audzijonyte, A" uniqKey="Audzijonyte A">A Audzijonyte</name></author><author><name sortKey="Ovcarenko, I" uniqKey="Ovcarenko I">I Ovcarenko</name></author><author><name sortKey="Bastrop, R" uniqKey="Bastrop R">R Bastrop</name></author><author><name sortKey="V Inol, R" uniqKey="V Inol R">R Väinölä</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Audouin, Jv" uniqKey="Audouin J">JV Audouin</name></author><author><name sortKey="Milne Edwards, H" uniqKey="Milne Edwards H">H Milne-Edwards</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Austin, Ad" uniqKey="Austin A">AD Austin</name></author><author><name sortKey="Dillon, N" uniqKey="Dillon N">N Dillon</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bakken, T" uniqKey="Bakken T">T Bakken</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Barnes, D" uniqKey="Barnes D">D Barnes</name></author><author><name sortKey="Fluke, C" uniqKey="Fluke C">C Fluke</name></author><author><name sortKey="Bourke, P" uniqKey="Bourke P">P Bourke</name></author><author><name sortKey="Parry, Ot" uniqKey="Parry O">OT Parry</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Barroso, R" uniqKey="Barroso R">R Barroso</name></author><author><name sortKey="Klautau, M" uniqKey="Klautau M">M Klautau</name></author><author><name sortKey="Sole Cava, Am" uniqKey="Sole Cava A">AM Solé-Cava</name></author><author><name sortKey="De Paiva, Pc" uniqKey="De Paiva P">PC de Paiva</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Benson, Da" uniqKey="Benson D">DA Benson</name></author><author><name sortKey="Karsch Mizrachi, I" uniqKey="Karsch Mizrachi I">I Karsch-Mizrachi</name></author><author><name sortKey="Lipman, Dj" uniqKey="Lipman D">DJ Lipman</name></author><author><name sortKey="Ostell, J" uniqKey="Ostell J">J Ostell</name></author><author><name sortKey="Wheeler, Dl" uniqKey="Wheeler D">DL Wheeler</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Berquist, Rm" uniqKey="Berquist R">RM Berquist</name></author><author><name sortKey="Gledhill, Km" uniqKey="Gledhill K">KM Gledhill</name></author><author><name sortKey="Peterson, Mw" uniqKey="Peterson M">MW Peterson</name></author><author><name sortKey="Doan, Ah" uniqKey="Doan A">AH Doan</name></author><author><name sortKey="Baxter, Gt" uniqKey="Baxter G">GT Baxter</name></author><author><name sortKey="Yopak, Ke" uniqKey="Yopak K">KE Yopak</name></author><author><name sortKey="Kang, N" uniqKey="Kang N">N Kang</name></author><author><name sortKey="Walker, Hj" uniqKey="Walker H">HJ Walker</name></author><author><name sortKey="Hastings, Pa" uniqKey="Hastings P">PA Hastings</name></author><author><name sortKey="Frank, Lr" uniqKey="Frank L">LR Frank</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Boistel, R" uniqKey="Boistel R">R Boistel</name></author><author><name sortKey="Swoger, J" uniqKey="Swoger J">J Swoger</name></author><author><name sortKey="Krzi, U" uniqKey="Krzi U">U Kržič</name></author><author><name sortKey="Fernandez, V" uniqKey="Fernandez V">V Fernandez</name></author><author><name sortKey="Gillet, B" uniqKey="Gillet B">B Gillet</name></author><author><name sortKey="Reynaud, Eg" uniqKey="Reynaud E">EG Reynaud</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bond, J" uniqKey="Bond J">J Bond</name></author><author><name sortKey="Hornschmeyer, T" uniqKey="Hornschmeyer T">T Hörnschmeyer</name></author><author><name sortKey="Young, P" uniqKey="Young P">P Young</name></author><author><name sortKey="Yang, L" uniqKey="Yang L">L Yang</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Briggs, Rt" uniqKey="Briggs R">RT Briggs</name></author><author><name sortKey="Chaffee, Je" uniqKey="Chaffee J">JE Chaffee</name></author><author><name sortKey="Anderson, M" uniqKey="Anderson M">M Anderson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Budd, Ge" uniqKey="Budd G">GE Budd</name></author><author><name sortKey="Olsson, L" uniqKey="Olsson L">L Olsson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Boggemann, M" uniqKey="Boggemann M">M Böggemann</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Carrera Parra, Lf" uniqKey="Carrera Parra L">LF Carrera-Parra</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Carrera Parra, Lf" uniqKey="Carrera Parra L">LF Carrera-Parra</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Carvalho, Mr" uniqKey="Carvalho M">MR Carvalho</name></author><author><name sortKey="Bockmann, Fa" uniqKey="Bockmann F">FA Bockmann</name></author><author><name sortKey="Amorim, Ds" uniqKey="Amorim D">DS Amorim</name></author><author><name sortKey="Brandao, Crf" uniqKey="Brandao C">CRF Brandão</name></author><author><name sortKey="Vivo, M" uniqKey="Vivo M">M Vivo</name></author><author><name sortKey="Figueiredo, Jl" uniqKey="Figueiredo J">JL Figueiredo</name></author><author><name sortKey="Britski, Ha" uniqKey="Britski H">HA Britski</name></author><author><name sortKey="Pinna, Mcc" uniqKey="Pinna M">MCC Pinna</name></author><author><name sortKey="Menezes, Na" uniqKey="Menezes N">NA Menezes</name></author><author><name sortKey="Marques, Fpl" uniqKey="Marques F">FPL Marques</name></author><author><name sortKey="Papavero, N" uniqKey="Papavero N">N Papavero</name></author><author><name sortKey="Cancello, Em" uniqKey="Cancello E">EM Cancello</name></author><author><name sortKey="Crisci, Jv" uniqKey="Crisci J">JV Crisci</name></author><author><name sortKey="Mceachran, Jd" uniqKey="Mceachran J">JD McEachran</name></author><author><name sortKey="Schelly, Rc" uniqKey="Schelly R">RC Schelly</name></author><author><name sortKey="Lundberg, Jg" uniqKey="Lundberg J">JG Lundberg</name></author><author><name sortKey="Gill, Ac" uniqKey="Gill A">AC Gill</name></author><author><name sortKey="Britz, R" uniqKey="Britz R">R Britz</name></author><author><name sortKey="Wheeler, Qd" uniqKey="Wheeler Q">QD Wheeler</name></author><author><name sortKey="Stiassny, Mlj" uniqKey="Stiassny M">MLJ Stiassny</name></author><author><name sortKey="Parenti, Lr" uniqKey="Parenti L">LR Parenti</name></author><author><name sortKey="Page, Lm" uniqKey="Page L">LM Page</name></author><author><name sortKey="Wheeler, Wc" uniqKey="Wheeler W">WC Wheeler</name></author><author><name sortKey="Faivovich, J" uniqKey="Faivovich J">J Faivovich</name></author><author><name sortKey="Vari, Rp" uniqKey="Vari R">RP Vari</name></author><author><name sortKey="Grande, L" uniqKey="Grande L">L Grande</name></author><author><name sortKey="Humphries, Cj" uniqKey="Humphries C">CJ Humphries</name></author><author><name sortKey="Desalle, R" uniqKey="Desalle R">R DeSalle</name></author><author><name sortKey="Ebach, Mc" uniqKey="Ebach M">MC Ebach</name></author><author><name sortKey="Nelson, Gj" uniqKey="Nelson G">GJ Nelson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Claparede, E" uniqKey="Claparede E">É Claparède</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Congote, J" uniqKey="Congote J">J Congote</name></author><author><name sortKey="Kabongo, L" uniqKey="Kabongo L">L Kabongo</name></author><author><name sortKey="Moreno, A" uniqKey="Moreno A">A Moreno</name></author><author><name sortKey="Segura, A" uniqKey="Segura A">A Segura</name></author><author><name sortKey="Beristain, A" uniqKey="Beristain A">A Beristain</name></author><author><name sortKey="Posada, J" uniqKey="Posada J">J Posada</name></author><author><name sortKey="Ruiz, O" uniqKey="Ruiz O">O Ruiz</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Congote, J" uniqKey="Congote J">J Congote</name></author><author><name sortKey="Kabongo, L" uniqKey="Kabongo L">L Kabongo</name></author><author><name sortKey="Moreno, A" uniqKey="Moreno A">A Moreno</name></author><author><name sortKey="Segura, A" uniqKey="Segura A">A Segura</name></author><author><name sortKey="Posada, J" uniqKey="Posada J">J Posada</name></author><author><name sortKey="Ruiz, O" uniqKey="Ruiz O">O Ruiz</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cook, Lg" uniqKey="Cook L">LG Cook</name></author><author><name sortKey="Edwards, Rd" uniqKey="Edwards R">RD Edwards</name></author><author><name sortKey="Crisp, Md" uniqKey="Crisp M">MD Crisp</name></author><author><name sortKey="Hardy, Nb" uniqKey="Hardy N">NB Hardy</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Csosz, S" uniqKey="Csosz S">S Csösz</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Deans, Ar" uniqKey="Deans A">AR Deans</name></author><author><name sortKey="Yoder, Mj" uniqKey="Yoder M">MJ Yoder</name></author><author><name sortKey="Balhoff, Jp" uniqKey="Balhoff J">JP Balhoff</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Deligiannidis, L" uniqKey="Deligiannidis L">L Deligiannidis</name></author><author><name sortKey="Jacob, Rjk" uniqKey="Jacob R">RJK Jacob</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dierick, M" uniqKey="Dierick M">M Dierick</name></author><author><name sortKey="Cnudde, V" uniqKey="Cnudde V">V Cnudde</name></author><author><name sortKey="Masschaele, B" uniqKey="Masschaele B">B Masschaele</name></author><author><name sortKey="Vlassenbroeck, J" uniqKey="Vlassenbroeck J">J Vlassenbroeck</name></author><author><name sortKey="Van Hoorebeke, L" uniqKey="Van Hoorebeke L">L Van Hoorebeke</name></author><author><name sortKey="Jacobs, P" uniqKey="Jacobs P">P Jacobs</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dinley, J" uniqKey="Dinley J">J Dinley</name></author><author><name sortKey="Hawkins, L" uniqKey="Hawkins L">L Hawkins</name></author><author><name sortKey="Paterson, Glj" uniqKey="Paterson G">GLJ Paterson</name></author><author><name sortKey="Ball, Ad" uniqKey="Ball A">AD Ball</name></author><author><name sortKey="Sinclair, I" uniqKey="Sinclair I">I Sinclair</name></author><author><name sortKey="Sinnett Jones, P" uniqKey="Sinnett Jones P">P Sinnett-Jones</name></author><author><name sortKey="Lanham, S" uniqKey="Lanham S">S Lanham</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dunlop, Ja" uniqKey="Dunlop J">JA Dunlop</name></author><author><name sortKey="Wirth, S" uniqKey="Wirth S">S Wirth</name></author><author><name sortKey="Penney, D" uniqKey="Penney D">D Penney</name></author><author><name sortKey="Mcneil, A" uniqKey="Mcneil A">A McNeil</name></author><author><name sortKey="Bradley, Rs" uniqKey="Bradley R">RS Bradley</name></author><author><name sortKey="Withers, Pj" uniqKey="Withers P">PJ Withers</name></author><author><name sortKey="Preziosi, Rf" uniqKey="Preziosi R">RF Preziosi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dunn, Cw" uniqKey="Dunn C">CW Dunn</name></author><author><name sortKey="Hejnol, A" uniqKey="Hejnol A">A Hejnol</name></author><author><name sortKey="Matus, Dq" uniqKey="Matus D">DQ Matus</name></author><author><name sortKey="Pang, K" uniqKey="Pang K">K Pang</name></author><author><name sortKey="Browne, We" uniqKey="Browne W">WE Browne</name></author><author><name sortKey="Smith, Sa" uniqKey="Smith S">SA Smith</name></author><author><name sortKey="Seaver, E" uniqKey="Seaver E">E Seaver</name></author><author><name sortKey="Rouse, Gw" uniqKey="Rouse G">GW Rouse</name></author><author><name sortKey="Obst, M" uniqKey="Obst M">M Obst</name></author><author><name sortKey="Edgecombe, Gd" uniqKey="Edgecombe G">GD Edgecombe</name></author><author><name sortKey="S Rensen, Mv" uniqKey="S Rensen M">MV Sørensen</name></author><author><name sortKey="Haddock, Shd" uniqKey="Haddock S">SHD Haddock</name></author><author><name sortKey="Schmidt Rhaesa, A" uniqKey="Schmidt Rhaesa A">A Schmidt-Rhaesa</name></author><author><name sortKey="Okusu, A" uniqKey="Okusu A">A Okusu</name></author><author><name sortKey="Kristensen, Rm" uniqKey="Kristensen R">RM Kristensen</name></author><author><name sortKey="Wheeler, Wc" uniqKey="Wheeler W">WC Wheeler</name></author><author><name sortKey="Martindale, Mq" uniqKey="Martindale M">MQ Martindale</name></author><author><name sortKey="Giribet, G" uniqKey="Giribet G">G Giribet</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Edgecombe, Gd" uniqKey="Edgecombe G">GD Edgecombe</name></author><author><name sortKey="Giribet, G" uniqKey="Giribet G">G Giribet</name></author><author><name sortKey="Dunn, Cw" uniqKey="Dunn C">CW Dunn</name></author><author><name sortKey="Hejnol, A" uniqKey="Hejnol A">A Hejnol</name></author><author><name sortKey="Kristensen, Rm" uniqKey="Kristensen R">RM Kristensen</name></author><author><name sortKey="Neves, Rc" uniqKey="Neves R">RC Neves</name></author><author><name sortKey="Rouse, Gw" uniqKey="Rouse G">GW Rouse</name></author><author><name sortKey="Worsaae, K" uniqKey="Worsaae K">K Worsaae</name></author><author><name sortKey="S Rensen, Mv" uniqKey="S Rensen M">MV Sørensen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Engel, K" uniqKey="Engel K">K Engel</name></author><author><name sortKey="Sommer, O" uniqKey="Sommer O">O Sommer</name></author><author><name sortKey="Ernst, C" uniqKey="Ernst C">C Ernst</name></author><author><name sortKey="Ertl, T" uniqKey="Ertl T">T Ertl</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Evenhuis, Nl" uniqKey="Evenhuis N">NL Evenhuis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Faulwetter, S" uniqKey="Faulwetter S">S Faulwetter</name></author><author><name sortKey="Dailianis, T" uniqKey="Dailianis T">T Dailianis</name></author><author><name sortKey="Vasileiadou, A" uniqKey="Vasileiadou A">A Vasileiadou</name></author><author><name sortKey="Arvanitidis, C" uniqKey="Arvanitidis C">C Arvanitidis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Folmer, O" uniqKey="Folmer O">O Folmer</name></author><author><name sortKey="Black, M" uniqKey="Black M">M Black</name></author><author><name sortKey="Hoeh, W" uniqKey="Hoeh W">W Hoeh</name></author><author><name sortKey="Lutz, R" uniqKey="Lutz R">R Lutz</name></author><author><name sortKey="Vrijenhoek, R" uniqKey="Vrijenhoek R">R Vrijenhoek</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="George, Jd" uniqKey="George J">JD George</name></author><author><name sortKey="Southward, Ec" uniqKey="Southward E">EC Southward</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Giribet, G" uniqKey="Giribet G">G Giribet</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Godfray, Hcj" uniqKey="Godfray H">HCJ Godfray</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Godfray, Hcj" uniqKey="Godfray H">HCJ Godfray</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Godfray, Hcj" uniqKey="Godfray H">HCJ Godfray</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Golding, Re" uniqKey="Golding R">RE Golding</name></author><author><name sortKey="Jones, As" uniqKey="Jones A">AS Jones</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Golding, Re" uniqKey="Golding R">RE Golding</name></author><author><name sortKey="Ponder, Wf" uniqKey="Ponder W">WF Ponder</name></author><author><name sortKey="Byrne, M" uniqKey="Byrne M">M Byrne</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gotherstrom, A" uniqKey="Gotherstrom A">A Götherstrom</name></author><author><name sortKey="Fischer, C" uniqKey="Fischer C">C Fischer</name></author><author><name sortKey="Linden, K" uniqKey="Linden K">K Linden</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Grieshaber, Bm" uniqKey="Grieshaber B">BM Grieshaber</name></author><author><name sortKey="Osborne, Dl" uniqKey="Osborne D">DL Osborne</name></author><author><name sortKey="Doubleday, Af" uniqKey="Doubleday A">AF Doubleday</name></author><author><name sortKey="Kaestle, Fa" uniqKey="Kaestle F">FA Kaestle</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Grube, Ae" uniqKey="Grube A">AE Grube</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Heim, I" uniqKey="Heim I">I Heim</name></author><author><name sortKey="Nickel, M" uniqKey="Nickel M">M Nickel</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hendrickx, H" uniqKey="Hendrickx H">H Hendrickx</name></author><author><name sortKey="Cnudde, V" uniqKey="Cnudde V">V Cnudde</name></author><author><name sortKey="Masschaele, B" uniqKey="Masschaele B">B Masschaele</name></author><author><name sortKey="Dierick, M" uniqKey="Dierick M">M Dierick</name></author><author><name sortKey="Vlassenbroeck, J" uniqKey="Vlassenbroeck J">J Vlassenbroeck</name></author><author><name sortKey="Van Hoorebeke, L" uniqKey="Van Hoorebeke L">L Van Hoorebeke</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Holford, P" uniqKey="Holford P">P Holford</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Huckstorf, K" uniqKey="Huckstorf K">K Huckstorf</name></author><author><name sortKey="Wirkner, Cs" uniqKey="Wirkner C">CS Wirkner</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="International, Commission On Zoological Nomenclature" uniqKey="International C">Commission on Zoological Nomenclature International</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Javidi, B" uniqKey="Javidi B">B Javidi</name></author><author><name sortKey="Tajahuerce, E" uniqKey="Tajahuerce E">E Tajahuerce</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Johnson, Pt" uniqKey="Johnson P">PT Johnson</name></author><author><name sortKey="Zimmerman, Sl" uniqKey="Zimmerman S">SL Zimmerman</name></author><author><name sortKey="Heath, D" uniqKey="Heath D">D Heath</name></author><author><name sortKey="Eng, J" uniqKey="Eng J">J Eng</name></author><author><name sortKey="Horton, Km" uniqKey="Horton K">KM Horton</name></author><author><name sortKey="Scott, Ww" uniqKey="Scott W">WW Scott</name></author><author><name sortKey="Fishman, Ek" uniqKey="Fishman E">EK Fishman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Joppa, Ln" uniqKey="Joppa L">LN Joppa</name></author><author><name sortKey="Roberts, Dl" uniqKey="Roberts D">DL Roberts</name></author><author><name sortKey="Pimm, Sl" uniqKey="Pimm S">SL Pimm</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Joppa, Ln" uniqKey="Joppa L">LN Joppa</name></author><author><name sortKey="Roberts, Dl" uniqKey="Roberts D">DL Roberts</name></author><author><name sortKey="Pimm, Sl" uniqKey="Pimm S">SL Pimm</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kaupp, G" uniqKey="Kaupp G">G Kaupp</name></author><author><name sortKey="Stepanenko, S" uniqKey="Stepanenko S">S Stepanenko</name></author><author><name sortKey="Herrmann, A" uniqKey="Herrmann A">A Herrmann</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kersemans, V" uniqKey="Kersemans V">V Kersemans</name></author><author><name sortKey="Thompson, J" uniqKey="Thompson J">J Thompson</name></author><author><name sortKey="Cornelissen, B" uniqKey="Cornelissen B">B Cornelissen</name></author><author><name sortKey="Woodcock, M" uniqKey="Woodcock M">M Woodcock</name></author><author><name sortKey="Allen, Pd" uniqKey="Allen P">PD Allen</name></author><author><name sortKey="Buls, N" uniqKey="Buls N">N Buls</name></author><author><name sortKey="Muschel, Rj" uniqKey="Muschel R">RJ Muschel</name></author><author><name sortKey="Hill, Ma" uniqKey="Hill M">MA Hill</name></author><author><name sortKey="Smart, Sc" uniqKey="Smart S">SC Smart</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Knapp, S" uniqKey="Knapp S">S Knapp</name></author><author><name sortKey="Bateman, Rm" uniqKey="Bateman R">RM Bateman</name></author><author><name sortKey="Chalmers, Nr" uniqKey="Chalmers N">NR Chalmers</name></author><author><name sortKey="Humphries, Cj" uniqKey="Humphries C">CJ Humphries</name></author><author><name sortKey="Rainbow, Ps" uniqKey="Rainbow P">PS Rainbow</name></author><author><name sortKey="Smith, Ab" uniqKey="Smith A">AB Smith</name></author><author><name sortKey="Taylor, Pd" uniqKey="Taylor P">PD Taylor</name></author><author><name sortKey="Vane Wright, Ri" uniqKey="Vane Wright R">RI Vane-Wright</name></author><author><name sortKey="Wilkinson, M" uniqKey="Wilkinson M">M Wilkinson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kumar, P" uniqKey="Kumar P">P Kumar</name></author><author><name sortKey="Ziegler, A" uniqKey="Ziegler A">A Ziegler</name></author><author><name sortKey="Grahn, A" uniqKey="Grahn A">A Grahn</name></author><author><name sortKey="Hee, Cs" uniqKey="Hee C">CS Hee</name></author><author><name sortKey="Ziegler, A" uniqKey="Ziegler A">A Ziegler</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Laforsch, C" uniqKey="Laforsch C">C Laforsch</name></author><author><name sortKey="Imhof, H" uniqKey="Imhof H">H Imhof</name></author><author><name sortKey="Sigl, R" uniqKey="Sigl R">R Sigl</name></author><author><name sortKey="Settles, M" uniqKey="Settles M">M Settles</name></author><author><name sortKey="He, M" uniqKey="He M">M Heß</name></author><author><name sortKey="Wanninger, A" uniqKey="Wanninger A">A Wanninger</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Laha, B" uniqKey="Laha B">B Laha</name></author><author><name sortKey="Sensharma, K" uniqKey="Sensharma K">K Sensharma</name></author><author><name sortKey="Schiffbauer, Jd" uniqKey="Schiffbauer J">JD Schiffbauer</name></author><author><name sortKey="Bowman, Da" uniqKey="Bowman D">DA Bowman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Leuckart, R" uniqKey="Leuckart R">R Leuckart</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Marxen, Jc" uniqKey="Marxen J">JC Marxen</name></author><author><name sortKey="Prymak, O" uniqKey="Prymak O">O Prymak</name></author><author><name sortKey="Beckmann, F" uniqKey="Beckmann F">F Beckmann</name></author><author><name sortKey="Neues, F" uniqKey="Neues F">F Neues</name></author><author><name sortKey="Epple, M" uniqKey="Epple M">M Epple</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mcpeek, Ma" uniqKey="Mcpeek M">MA McPeek</name></author><author><name sortKey="Symes, Lb" uniqKey="Symes L">LB Symes</name></author><author><name sortKey="Zong, Dm" uniqKey="Zong D">DM Zong</name></author><author><name sortKey="Mcpeek, Cl" uniqKey="Mcpeek C">CL McPeek</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Metscher, Bd" uniqKey="Metscher B">BD Metscher</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Metscher, Bd" uniqKey="Metscher B">BD Metscher</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Midgley, Pa" uniqKey="Midgley P">PA Midgley</name></author><author><name sortKey="Dunin Borkowski, Re" uniqKey="Dunin Borkowski R">RE Dunin-Borkowski</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mildenberger, P" uniqKey="Mildenberger P">P Mildenberger</name></author><author><name sortKey="Jensch, P" uniqKey="Jensch P">P Jensch</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Molineux, A" uniqKey="Molineux A">A Molineux</name></author><author><name sortKey="Scott, Rw" uniqKey="Scott R">RW Scott</name></author><author><name sortKey="Ketcham, Ra" uniqKey="Ketcham R">RA Ketcham</name></author><author><name sortKey="Maisano, Ja" uniqKey="Maisano J">JA Maisano</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Nickel, M" uniqKey="Nickel M">M Nickel</name></author><author><name sortKey="Donath, T" uniqKey="Donath T">T Donath</name></author><author><name sortKey="Schweikert, M" uniqKey="Schweikert M">M Schweikert</name></author><author><name sortKey="Beckmann, F" uniqKey="Beckmann F">F Beckmann</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Nygren, A" uniqKey="Nygren A">A Nygren</name></author><author><name sortKey="Eklof, J" uniqKey="Eklof J">J Eklöf</name></author><author><name sortKey="Pleijel, F" uniqKey="Pleijel F">F Pleijel</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Nygren, A" uniqKey="Nygren A">A Nygren</name></author><author><name sortKey="Pleijel, F" uniqKey="Pleijel F">F Pleijel</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Osborn, Kj" uniqKey="Osborn K">KJ Osborn</name></author><author><name sortKey="Rouse, Gw" uniqKey="Rouse G">GW Rouse</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Pallas, Ps" uniqKey="Pallas P">PS Pallas</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Paredes, Um" uniqKey="Paredes U">UM Paredes</name></author><author><name sortKey="Prys Jones, R" uniqKey="Prys Jones R">R Prys-Jones</name></author><author><name sortKey="Adams, M" uniqKey="Adams M">M Adams</name></author><author><name sortKey="Groombridge, J" uniqKey="Groombridge J">J Groombridge</name></author><author><name sortKey="Kundu, S" uniqKey="Kundu S">S Kundu</name></author><author><name sortKey="Agapow, P M" uniqKey="Agapow P">P-M Agapow</name></author><author><name sortKey="Abel, Rl" uniqKey="Abel R">RL Abel</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Patek, Sn" uniqKey="Patek S">SN Patek</name></author><author><name sortKey="Nowroozi, Bn" uniqKey="Nowroozi B">BN Nowroozi</name></author><author><name sortKey="Baio, Je" uniqKey="Baio J">JE Baio</name></author><author><name sortKey="Caldwell, Rl" uniqKey="Caldwell R">RL Caldwell</name></author><author><name sortKey="Summers, Ap" uniqKey="Summers A">AP Summers</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Penev, L" uniqKey="Penev L">L Penev</name></author><author><name sortKey="Agosti, D" uniqKey="Agosti D">D Agosti</name></author><author><name sortKey="Georgiev, T" uniqKey="Georgiev T">T Georgiev</name></author><author><name sortKey="Catapano, T" uniqKey="Catapano T">T Catapano</name></author><author><name sortKey="Miller, J" uniqKey="Miller J">J Miller</name></author><author><name sortKey="Blagoderov, V" uniqKey="Blagoderov V">V Blagoderov</name></author><author><name sortKey="Roberts, D" uniqKey="Roberts D">D Roberts</name></author><author><name sortKey="Smith, Vs" uniqKey="Smith V">VS Smith</name></author><author><name sortKey="Brake, I" uniqKey="Brake I">I Brake</name></author><author><name sortKey="Ryrcroft, S" uniqKey="Ryrcroft S">S Ryrcroft</name></author><author><name sortKey="Scott, B" uniqKey="Scott B">B Scott</name></author><author><name sortKey="Johnson, Nf" uniqKey="Johnson N">NF Johnson</name></author><author><name sortKey="Morris, R A" uniqKey="Morris R">R a. Morris</name></author><author><name sortKey="Sautter, G" uniqKey="Sautter G">G Sautter</name></author><author><name sortKey="Chavan, V" uniqKey="Chavan V">V Chavan</name></author><author><name sortKey="Robertson, T" uniqKey="Robertson T">T Robertson</name></author><author><name sortKey="Remsen, D" uniqKey="Remsen D">D Remsen</name></author><author><name sortKey="Stoev, P" uniqKey="Stoev P">P Stoev</name></author><author><name sortKey="Parr, C" uniqKey="Parr C">C Parr</name></author><author><name sortKey="Knapp, S" uniqKey="Knapp S">S Knapp</name></author><author><name sortKey="Kress, Wj" uniqKey="Kress W">WJ Kress</name></author><author><name sortKey="Thompson, Fc" uniqKey="Thompson F">FC Thompson</name></author><author><name sortKey="Erwin, T" uniqKey="Erwin T">T Erwin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Penney, D" uniqKey="Penney D">D Penney</name></author><author><name sortKey="Dierick, M" uniqKey="Dierick M">M Dierick</name></author><author><name sortKey="Cnudde, V" uniqKey="Cnudde V">V Cnudde</name></author><author><name sortKey="Masschaele, B" uniqKey="Masschaele B">B Masschaele</name></author><author><name sortKey="Vlassenbroeck, J" uniqKey="Vlassenbroeck J">J Vlassenbroeck</name></author><author><name sortKey="Van Hoorebeke, L" uniqKey="Van Hoorebeke L">L Van Hoorebeke,</name></author><author><name sortKey="Jacobs, P" uniqKey="Jacobs P">P Jacobs</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Philippe, H" uniqKey="Philippe H">H Philippe</name></author><author><name sortKey="Derelle, R" uniqKey="Derelle R">R Derelle</name></author><author><name sortKey="Lopez, P" uniqKey="Lopez P">P Lopez</name></author><author><name sortKey="Pick, K" uniqKey="Pick K">K Pick</name></author><author><name sortKey="Borchiellini, C" uniqKey="Borchiellini C">C Borchiellini</name></author><author><name sortKey="Boury Esnault, N" uniqKey="Boury Esnault N">N Boury-Esnault</name></author><author><name sortKey="Vacelet, J" uniqKey="Vacelet J">J Vacelet</name></author><author><name sortKey="Renard, E" uniqKey="Renard E">E Renard</name></author><author><name sortKey="Houliston, E" uniqKey="Houliston E">E Houliston</name></author><author><name sortKey="Queinnec, E" uniqKey="Queinnec E">E Quéinnec</name></author><author><name sortKey="Da, Silva C" uniqKey="Da S">Silva C Da</name></author><author><name sortKey="Wincker, P" uniqKey="Wincker P">P Wincker</name></author><author><name sortKey="Le Guyader, H" uniqKey="Le Guyader H">H Le Guyader</name></author><author><name sortKey="Leys, S" uniqKey="Leys S">S Leys</name></author><author><name sortKey="Jackson, Dj" uniqKey="Jackson D">DJ Jackson</name></author><author><name sortKey="Schreiber, F" uniqKey="Schreiber F">F Schreiber</name></author><author><name sortKey="Erpenbeck, D" uniqKey="Erpenbeck D">D Erpenbeck</name></author><author><name sortKey="Morgenstern, B" uniqKey="Morgenstern B">B Morgenstern</name></author><author><name sortKey="Worheide, G" uniqKey="Worheide G">G Wörheide</name></author><author><name sortKey="Manuel, M" uniqKey="Manuel M">M Manuel</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Pianykh, Os" uniqKey="Pianykh O">OS Pianykh</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Postnov, A" uniqKey="Postnov A">A Postnov</name></author><author><name sortKey="De Clerck, N" uniqKey="De Clerck N">N de Clerck</name></author><author><name sortKey="Sasov, A" uniqKey="Sasov A">A Sasov</name></author><author><name sortKey="Van Dyck, D" uniqKey="Van Dyck D">D Van Dyck</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Prohaska, S" uniqKey="Prohaska S">S Prohaska</name></author><author><name sortKey="Hutanu, A" uniqKey="Hutanu A">A Hutanu</name></author><author><name sortKey="Hege, H C" uniqKey="Hege H">H-C Hege</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Puce, S" uniqKey="Puce S">S Puce</name></author><author><name sortKey="Pica, D" uniqKey="Pica D">D Pica</name></author><author><name sortKey="Brun, F" uniqKey="Brun F">F Brun</name></author><author><name sortKey="Mancini, L" uniqKey="Mancini L">L Mancini</name></author><author><name sortKey="Bavestrello, G" uniqKey="Bavestrello G">G Bavestrello</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Qiong, X" uniqKey="Qiong X">X Qiong</name></author><author><name sortKey="Hengyong, Y" uniqKey="Hengyong Y">Y Hengyong</name></author><author><name sortKey="Bennet, J" uniqKey="Bennet J">J Bennet</name></author><author><name sortKey="Peng, H" uniqKey="Peng H">H Peng</name></author><author><name sortKey="Zainon, R" uniqKey="Zainon R">R Zainon</name></author><author><name sortKey="Doesburg, R" uniqKey="Doesburg R">R Doesburg</name></author><author><name sortKey="Opiee, A" uniqKey="Opiee A">A OpieE</name></author><author><name sortKey="Walsh, M" uniqKey="Walsh M">M Walsh</name></author><author><name sortKey="Haiou, S" uniqKey="Haiou S">S Haiou</name></author><author><name sortKey="Butler, A" uniqKey="Butler A">A Butler</name></author><author><name sortKey="Butler, P" uniqKey="Butler P">P Butler</name></author><author><name sortKey="Xuanqin, M" uniqKey="Xuanqin M">M Xuanqin</name></author><author><name sortKey="Ge, W" uniqKey="Ge W">W Ge</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Quatrefages, A" uniqKey="Quatrefages A">A Quatrefages</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Quintarelli, G" uniqKey="Quintarelli G">G Quintarelli</name></author><author><name sortKey="Bellocci, M" uniqKey="Bellocci M">M Bellocci</name></author><author><name sortKey="Geremia, R" uniqKey="Geremia R">R Geremia</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Rowe, T" uniqKey="Rowe T">T Rowe</name></author><author><name sortKey="Frank, Lr" uniqKey="Frank L">LR Frank</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ruthensteiner, B" uniqKey="Ruthensteiner B">B Ruthensteiner</name></author><author><name sortKey="Baeumler, N" uniqKey="Baeumler N">N Baeumler</name></author><author><name sortKey="Barnes, Dg" uniqKey="Barnes D">DG Barnes</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ruthensteiner, B" uniqKey="Ruthensteiner B">B Ruthensteiner</name></author><author><name sortKey="Hess, M" uniqKey="Hess M">M Hess</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="San Martin, G" uniqKey="San Martin G">G San Martín</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Smith, V" uniqKey="Smith V">V Smith</name></author><author><name sortKey="Blagoderov, V" uniqKey="Blagoderov V">V Blagoderov</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sutton, Md" uniqKey="Sutton M">MD Sutton</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sutton, Md" uniqKey="Sutton M">MD Sutton</name></author><author><name sortKey="Garwood, Rj" uniqKey="Garwood R">RJ Garwood</name></author><author><name sortKey="Siveter, Dj" uniqKey="Siveter D">DJ Siveter</name></author><author><name sortKey="Siveter, Dj" uniqKey="Siveter D">DJ Siveter</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tamura, K" uniqKey="Tamura K">K Tamura</name></author><author><name sortKey="Peterson, D" uniqKey="Peterson D">D Peterson</name></author><author><name sortKey="Peterson, N" uniqKey="Peterson N">N Peterson</name></author><author><name sortKey="Stecher, G" uniqKey="Stecher G">G Stecher</name></author><author><name sortKey="Nei, M" uniqKey="Nei M">M Nei</name></author><author><name sortKey="Kumar, S" uniqKey="Kumar S">S Kumar</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wang, G" uniqKey="Wang G">G Wang</name></author><author><name sortKey="Yu, H" uniqKey="Yu H">H Yu</name></author><author><name sortKey="De Man, B" uniqKey="De Man B">B De Man</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Westheide, W" uniqKey="Westheide W">W Westheide</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wheeler, Qd" uniqKey="Wheeler Q">QD Wheeler</name></author><author><name sortKey="Raven, Ph" uniqKey="Raven P">PH Raven</name></author><author><name sortKey="Wilson, Eo" uniqKey="Wilson E">EO Wilson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wheeler, Q" uniqKey="Wheeler Q">Q Wheeler</name></author><author><name sortKey="Thierry, B" uniqKey="Thierry B">B Thierry</name></author><author><name sortKey="Coddington, J" uniqKey="Coddington J">J Coddington</name></author><author><name sortKey="Gostony, T" uniqKey="Gostony T">T Gostony</name></author><author><name sortKey="Hamilton, A" uniqKey="Hamilton A">A Hamilton</name></author><author><name sortKey="Larimer, R" uniqKey="Larimer R">R Larimer</name></author><author><name sortKey="Polaszek, A" uniqKey="Polaszek A">A Polaszek</name></author><author><name sortKey="Schauff, M" uniqKey="Schauff M">M Schauff</name></author><author><name sortKey="Solis, A" uniqKey="Solis A">A Solis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wilhelm, G" uniqKey="Wilhelm G">G Wilhelm</name></author><author><name sortKey="Handschuh, S" uniqKey="Handschuh S">S Handschuh</name></author><author><name sortKey="Plant, J" uniqKey="Plant J">J Plant</name></author><author><name sortKey="Nemeschkal, Hl" uniqKey="Nemeschkal H">HL Nemeschkal</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wirkner, Cs" uniqKey="Wirkner C">CS Wirkner</name></author><author><name sortKey="Prendini, L" uniqKey="Prendini L">L Prendini</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wolff, S" uniqKey="Wolff S">S Wolff</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zanol, J" uniqKey="Zanol J">J Zanol</name></author><author><name sortKey="Bettoso, N" uniqKey="Bettoso N">N Bettoso</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zanol, J" uniqKey="Zanol J">J Zanol</name></author><author><name sortKey="Halanych, Km" uniqKey="Halanych K">KM Halanych</name></author><author><name sortKey="Struck, Th" uniqKey="Struck T">TH Struck</name></author><author><name sortKey="Fauchald, K" uniqKey="Fauchald K">K Fauchald</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ziegler, A" uniqKey="Ziegler A">A Ziegler</name></author><author><name sortKey="Faber, C" uniqKey="Faber C">C Faber</name></author><author><name sortKey="Mueller, S" uniqKey="Mueller S">S Mueller</name></author><author><name sortKey="Bartolomaeus, T" uniqKey="Bartolomaeus T">T Bartolomaeus</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ziegler, A" uniqKey="Ziegler A">A Ziegler</name></author><author><name sortKey="Kunth, M" uniqKey="Kunth M">M Kunth</name></author><author><name sortKey="Mueller, S" uniqKey="Mueller S">S Mueller</name></author><author><name sortKey="Bock, C" uniqKey="Bock C">C Bock</name></author><author><name sortKey="Pohmann, R" uniqKey="Pohmann R">R Pohmann</name></author><author><name sortKey="Schroder, L" uniqKey="Schroder L">L Schröder</name></author><author><name sortKey="Faber, C" uniqKey="Faber C">C Faber</name></author><author><name sortKey="Giribet, G" uniqKey="Giribet G">G Giribet</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ziegler, A" uniqKey="Ziegler A">A Ziegler</name></author><author><name sortKey="Mietchen, D" uniqKey="Mietchen D">D Mietchen</name></author><author><name sortKey="Faber, C" uniqKey="Faber C">C Faber</name></author><author><name sortKey="Von Hausen, W" uniqKey="Von Hausen W">W von Hausen</name></author><author><name sortKey="Schobel, C" uniqKey="Schobel C">C Schöbel</name></author><author><name sortKey="Sellerer, M" uniqKey="Sellerer M">M Sellerer</name></author><author><name sortKey="Ziegler, A" uniqKey="Ziegler A">A Ziegler</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ziegler, A" uniqKey="Ziegler A">A Ziegler</name></author><author><name sortKey="Ogurreck, M" uniqKey="Ogurreck M">M Ogurreck</name></author><author><name sortKey="Steinke, T" uniqKey="Steinke T">T Steinke</name></author><author><name sortKey="Beckmann, F" uniqKey="Beckmann F">F Beckmann</name></author><author><name sortKey="Prohaska, S" uniqKey="Prohaska S">S Prohaska</name></author><author><name sortKey="Ziegler, A" uniqKey="Ziegler A">A Ziegler</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zimmermann, D" uniqKey="Zimmermann D">D Zimmermann</name></author><author><name sortKey="Randolf, S" uniqKey="Randolf S">S Randolf</name></author><author><name sortKey="Metscher, Bd" uniqKey="Metscher B">BD Metscher</name></author><author><name sortKey="Aspock, U" uniqKey="Aspock U">U Aspöck</name></author></analytic></biblStruct></listBibl></div1></back></TEI><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Zookeys</journal-id><journal-id journal-id-type="iso-abbrev">Zookeys</journal-id><journal-id journal-id-type="publisher-id">ZooKeys</journal-id><journal-title-group><journal-title>ZooKeys</journal-title></journal-title-group><issn pub-type="ppub">1313-2989</issn><issn pub-type="epub">1313-2970</issn><publisher><publisher-name>Pensoft Publishers</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">23653515</article-id><article-id pub-id-type="pmc">3591762</article-id><article-id pub-id-type="doi">10.3897/zookeys.263.4261</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Micro-computed tomography: Introducing new dimensions to taxonomy</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Faulwetter</surname><given-names>Sarah</given-names></name><xref ref-type="aff" rid="A1">1</xref><xref ref-type="aff" rid="A2">2</xref></contrib><contrib contrib-type="author"><name><surname>Vasileiadou</surname><given-names>Aikaterini</given-names></name><xref ref-type="aff" rid="A2">2</xref><xref ref-type="aff" rid="A3">3</xref></contrib><contrib contrib-type="author"><name><surname>Kouratoras</surname><given-names>Michail</given-names></name><xref ref-type="aff" rid="A4">4</xref></contrib><contrib contrib-type="author"><name><surname>Thanos Dailianis</surname><given-names></given-names></name><xref ref-type="aff" rid="A2">2</xref></contrib><contrib contrib-type="author"><name><surname>Arvanitidis</surname><given-names> Christos</given-names></name><xref ref-type="aff" rid="A2">2</xref></contrib></contrib-group><aff id="A1"><label>1</label>Department of Zoology-Marine Biology, Faculty of Biology, National and Kapodestrian University of Athens, Panepistimiopolis, 15784, Athens, Greece,</aff><aff id="A2"><label>2</label>Institute for Marine Biology, Biotechnology and Aquaculture, Hellenic Centre for Marine Research, 71003 Heraklion, Crete, Greece</aff><aff id="A3"><label>3</label>Department of Biology, University of Patras, 26504, Rio, Patras, Greece</aff><aff id="A4"><label>4</label>Hellenic Centre for Marine Research, 71003 Heraklion, Crete, Greece</aff><author-notes><corresp>Corresponding author: Sarah Faulwetter (<email xlink:type="simple">sarifa@hcmr.gr</email>)</corresp><fn fn-type="edited-by"><p>Academic editor: V. Smith</p></fn></author-notes><pub-date pub-type="collection"><year>2013</year></pub-date><pub-date pub-type="epub"><day>4</day><month>2</month><year>2013</year></pub-date><issue>263</issue><fpage>1</fpage><lpage>45</lpage><history><date date-type="received"><day>7</day><month>11</month><year>2012</year></date><date date-type="accepted"><day>28</day><month>1</month><year>2013</year></date></history><permissions><copyright-statement>Sarah Faulwetter, Aikaterini Vasileiadou, Michail Kouratoras, Thanos Dailianis, Christos Arvanitidis</copyright-statement><license license-type="creative-commons-attribution" xlink:href="http://creativecommons.org/licenses/by/3.0"><license-p>This is an open access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC-BY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p></license></permissions><abstract><label>Abstract</label><p>Continuous improvements in the resolution of three-dimensional imaging have led to an increased application of these techniques in conventional taxonomic research in recent years. Coupled with an ever increasing research effort in cybertaxonomy, three-dimensional imaging could give a boost to the development of virtual specimen collections, allowing rapid and simultaneous access to accurate virtual representations of type material. This paper explores the potential of micro-computed tomography (X-ray micro-tomography), a non-destructive three-dimensional imaging technique based on mapping X-ray attenuation in the scanned object, for supporting research in systematics and taxonomy. The subsequent use of these data as virtual type material, so-called “cybertypes”, and the creation of virtual collections lie at the core of this potential. Sample preparation, image acquisition, data processing and presentation of results are demonstrated using polychaetes (bristle worms), a representative taxon of macro-invertebrates, as a study object. Effects of the technique on the morphological, anatomical and molecular identity of the specimens are investigated. The paper evaluates the results and discusses the potential and the limitations of the technique for creating cybertypes. It also discusses the challenges that the community might face to establish virtual collections. Potential future applications of three-dimensional information in taxonomic research are outlined, including an outlook to new ways of producing, disseminating and publishing taxonomic information.</p></abstract><kwd-group><label>Keywords</label><kwd>Micro-computed tomography</kwd><kwd>systematics</kwd><kwd>taxonomy</kwd><kwd>3D</kwd><kwd>visualisation</kwd><kwd>interactive PDF</kwd><kwd>polychaetes</kwd><kwd>cybertypes</kwd><kwd>cybertaxonomy</kwd><pmc-comment>PageBreak</pmc-comment>
</kwd-group></article-meta></front><body><sec><title>Introduction</title><p>Morphology-based taxonomy has been at the heart of systematic research for over two centuries. Over the last decades, however, the dominant role of morphology in systematics and phylogenetics has been challenged by an increasing number of analyses supported by molecular data (<xref ref-type="bibr" rid="B22">Cook et al. 2010</xref>, <xref ref-type="bibr" rid="B36">Giribet 2010</xref>). Sequence data are being produced at a rapid speed and are readily available for constructing phylogenies or delimiting species. However, the formal description and naming of species and other biological units is still central to biodiversity research (<xref ref-type="bibr" rid="B14">Budd and Olsson 2007</xref>, <xref ref-type="bibr" rid="B24">Deans et al. 2011</xref>). This highly accelerated data acquisition creates an imbalance between availability of data and the human power to actually interpret them and thus to create new knowledge. In taxonomy, this “gap in scalability” (<xref ref-type="bibr" rid="B36">Giribet 2010</xref>) becomes even more problematic due to the time-consuming and still largely manual process of describing and naming new species which cannot keep up with the speed at which new information becomes available from the molecular world. As a consequence, a steadily increasing number of species are delimited genetically but lack a formal, morphology-based description (e.g. <xref ref-type="bibr" rid="B3">Audzijonyte et al. 2008</xref>, <xref ref-type="bibr" rid="B8">Barroso et al. 2009</xref>, <xref ref-type="bibr" rid="B71">Osborn and Rouse 2010</xref>). This problem is amplified by the so-called “taxonomic impediment”: fewer and fewer resources—both human and financial—are available for conventional taxonomic and systematic research (<xref ref-type="bibr" rid="B18">Carvalho et al. 2007</xref>), and the field is lacking a global electronic infrastructure (<xref ref-type="bibr" rid="B37">Godfray 2002a</xref>, <xref ref-type="bibr" rid="B95">Wheeler et al. 2004</xref>). The controversial debate over possible causes and remedies for the stagnation the discipline is experiencing (e.g. <xref ref-type="bibr" rid="B56">Knapp et al. 2002</xref>, <xref ref-type="bibr" rid="B38">Godfray 2002b</xref>, <xref ref-type="bibr" rid="B18">Carvalho et al. 2007</xref>, <xref ref-type="bibr" rid="B32">Evenhuis 2007</xref>, <xref ref-type="bibr" rid="B52">Joppa et al. 2011a</xref>, <xref ref-type="bibr" rid="B53">b</xref>) has stimulated governments and funding agencies to increasingly recognise its importance, and several encouraging developments have arisen over the last years. Besides releasing funds for training and education, much funding has been allocated to the field of cybertaxonomy, allowing the development of internet-based tools and resources aiming to boost taxonomic research and to accelerate the process of new species descriptions and systematic analyses. These developments include online resources such as name-based registers (e.g. Catalogue of Life – <ext-link ext-link-type="uri" xlink:href="http://www.catalogueoflife.org">http://www.catalogueoflife.org</ext-link> , the Global Names Architecture – <ext-link ext-link-type="uri" xlink:href="http://www.globalnames.org">http://www.globalnames.org</ext-link> , ZooBank – <ext-link ext-link-type="uri" xlink:href="http://www.zoobank.org">http://www.zoobank.org</ext-link> , the World Register of Marine Species – <ext-link ext-link-type="uri" xlink:href="http://www.marinespecies.org">http://www.marinespecies.org</ext-link> ), biogeographic databases (e.g. the Ocean Biogeographic Information System – <ext-link ext-link-type="uri" xlink:href="http://www.iobis.org">http://www.iobis.org</ext-link> , the Global Biodiversity Information Facility – <ext-link ext-link-type="uri" xlink:href="http://www.gbif.org">http://www.gbif.org</ext-link> ), aggregators and curators (e.g. Fishbase –<ext-link ext-link-type="uri" xlink:href="http://www.fishbase.org">http://www.fishbase.org</ext-link> , Encyclopedia of Life – <ext-link ext-link-type="uri" xlink:href="http://www.eol.org">http://www.eol.org</ext-link> ), virtual research environments targeted towards taxonomy (Scratchpads – <ext-link ext-link-type="uri" xlink:href="http://www.scratchpads.eu">http://www.scratchpads.eu</ext-link> ), increasing availability of literature, both for legacy literature (Biodiversity Heritage Library – <ext-link ext-link-type="uri" xlink:href="http://www.biodiversitylibrary.org">http://www.biodiversitylibrary.org</ext-link> ) as well as an increase in open access literature and journals aiming at rapid publication of taxonomic treatments, including new publication models such as semantically enhanced information (<xref ref-type="bibr" rid="B75">Penev et al. 2010</xref>).</p><p>Despite the increase in information availability through these initiatives, one of the main bottlenecks in conventional taxonomy and systematics is still the availability of type material and thus reliable morphological information (<xref ref-type="bibr" rid="B39">Godfray 2007</xref>, <xref ref-type="bibr" rid="B96">Wheeler et al. 2012</xref>). Loans from museums are often difficult or even impossible to obtain and the time and effort to gather the material needed for a systematic revision can take months or years. The tediousness of this work contributes even further to the stagnation of the discipline: new data are produced at a slow pace, and this lack of new, readily accessible and computer-retrievable morphological data (<xref ref-type="bibr" rid="B24">Deans et al. 2011</xref>) prevents the testing of large-scale hypotheses, as they currently become common in phylogenomic analyses (<xref ref-type="bibr" rid="B29">Dunn et al. 2008</xref>, <xref ref-type="bibr" rid="B77">Philippe et al. 2009</xref>, <xref ref-type="bibr" rid="B30">Edgecombe et al. 2011</xref>). <xref ref-type="bibr" rid="B39">Godfray (2007)</xref> has argued that a possible solution to overcoming this bottleneck could be a web-based taxonomy using “cybertypes”, “based on the very best current imaging methods” (<xref ref-type="bibr" rid="B39">Godfray 2007</xref>), an idea immediately criticised by others (e.g <xref ref-type="bibr" rid="B18">Carvalho et al. 2007</xref>). The idea of creating virtual collections of taxonomic material is, however, indeed enticing, and first implementations of accurate imaging methods, mass digitisations and remote access to digital material have been recently presented in a dedicated collection of papers (<xref ref-type="bibr" rid="B89">Smith and Blagoderov 2012</xref>, and references therein). Technological advances and a new generation of imaging techniques will inevitably open new horizons not only by providing rapid access to first-hand morphological information but also by making this information accessible to humans and computers alike. Non-destructive three-dimensional imaging techniques such as confocal laser scanning microscopy (cLSM), optical projection tomography (OPT), magnetic resonance imaging (MRI) and micro-computed tomography (micro-CT), allow for rapid creation of high-resolution morphological and anatomical data in three dimensions (<xref ref-type="bibr" rid="B36">Giribet 2010</xref>, <xref ref-type="bibr" rid="B102">Ziegler et al. 2008</xref>, <xref ref-type="bibr" rid="B105">2010</xref>, 2011a, for a detailed overview of the different techniques see e.g. <xref ref-type="bibr" rid="B11">Boistel et al. 2011</xref>, <xref ref-type="bibr" rid="B58">Laforsch et al. 2012</xref>). These techniques allow detailed virtual reconstructions of the morphology and anatomy of specimens and subsequent interactive manipulation (e.g. rotation, virtual dissection) and analysis of these data. Not only are they ideal for the digitisation of taxonomically important morphological information, but they allow new kinds of analyses (e.g. morphometrics in three dimensions) to be performed, thus creating novel directions of research. Indeed, the ability of these technologies to create three-dimensional, interactive models with a resolution in the micrometre scale or even below, combined with non-destructive sample assessment (as opposed to three-dimensional reconstruction of histological sections), has lately started to attract the attention of researchers beyond the traditional (clinical) applications of the methods, boosted by an increasing accessibility of micro-CT scanners and rapid computational advances. Particularly, invertebrate zoologists have started to employ micro-MRI (for an overview of taxa imaged so far with MRI see <xref ref-type="bibr" rid="B103">Ziegler et al. 2011a</xref>) and micro-CT. Several studies already show the potential of these methods to deliver new data to test taxonomic hypotheses (<xref ref-type="bibr" rid="B45">Heim and Nickel 2010</xref>, <xref ref-type="bibr" rid="B62">McPeek et al. 2011</xref>, <xref ref-type="bibr" rid="B23">Csösz 2012</xref>). They also provide new insights into morphology and anatomy (<xref ref-type="bibr" rid="B40">Golding and Jones 2006</xref>, <xref ref-type="bibr" rid="B47">Holford 2008</xref>, <xref ref-type="bibr" rid="B27">Dinley et al. 2010</xref>, <xref ref-type="bibr" rid="B48">Huckstorf and Wirkner 2011</xref>), functional morphology (<xref ref-type="bibr" rid="B2">Alba-Tercedor and Sánchez-Tocino 2011</xref>, <xref ref-type="bibr" rid="B12">Bond et al. 2008</xref>, <xref ref-type="bibr" rid="B68">Nickel et al. 2006</xref>, <xref ref-type="bibr" rid="B74">Patek et al. 2007</xref>, <xref ref-type="bibr" rid="B97">Wilhelm et al. 2011</xref>) and developmental studies (<xref ref-type="bibr" rid="B79">Postnov et al. 2002</xref>, <xref ref-type="bibr" rid="B61">Marxen et al. 2007</xref>, <xref ref-type="bibr" rid="B81">Puce et al. 2012</xref>) by studying species through a virtual, three-dimensional model. In palaeobiology, the <pmc-comment>PageBreak</pmc-comment>technique is, for example, frequently used to reveal the morphology and even anatomy of fossilised organisms that cannot be removed from their enclosure medium (<xref ref-type="bibr" rid="B26">Dierick et al. 2007</xref>, <xref ref-type="bibr" rid="B28">Dunlop et al. 2011</xref>, <xref ref-type="bibr" rid="B46">Hendrickx et al. 2006</xref>, <xref ref-type="bibr" rid="B67">Molineux et al. 2007</xref>, <xref ref-type="bibr" rid="B76">Penney et al. 2007</xref>, <xref ref-type="bibr" rid="B90">Sutton 2008</xref>). Most of these studies have imaged few or a single specimens, but some have harnessed the power of non-invasive, three-dimensional imaging to create vast amounts of data for large-scale systematic analyses (<xref ref-type="bibr" rid="B98">Wirkner and Prendini 2007</xref>, <xref ref-type="bibr" rid="B102">Ziegler et al. 2008</xref>, <xref ref-type="bibr" rid="B41">Golding et al. 2009</xref>, <xref ref-type="bibr" rid="B106">Zimmermann et al. 2011</xref>).</p><p>Despite the increasing use of these new imaging methods, most of the recently created datasets might not qualify for the notion of a cybertype. In most studies, specimens were prepared and imaged with a specific hypothesis in mind, focusing on certain morphological characteristics and omitting others, and the resulting data might thus not be useful for other purposes. Datasets that are intended to serve as a cybertype should fulfil at least the following three basic assumptions: (a) A cybertype should provide morphological and anatomical information of the same accuracy and reliability as provided by the physical type material, independently of a specific research question in mind; (b) A cybertype should be linked to the original type material, which can be consulted if in doubt. This implies that any method used to create the cybertype should not affect the morphological, anatomical and molecular identity of the original specimen (e.g. holotype, paratype or neotype); (c) A cybertype has to be retrievable and freely accessible. This involves making the data available through a reliable (internet) source under an open access licence and providing adequate security measures, such as archiving, backups and ensuring data format compatibility in the future, and allowing the annotation of the dataset with metadata in order to be retrievable and interpretable.</p><p>Towards this end, this study explores the potential of micro-computed tomography to create high-throughput morphological and anatomical data to support systematic and taxonomic studies by using polychaetes (bristle worms) as a demonstration taxon for macro-invertebrates. This taxon has been chosen because of the diversity of shapes and tissue types occurring among its members, allowing the investigation of the behaviour of the methodology across a range of samples with different characteristics. The outcomes are evaluated with regard to the first requirement for constituting a potential cybertype, that is, their ability to deliver reliable information on diagnostic and systematically important characters. However, from sample preparation to the final presentation of the results many steps are involved which may affect both the outcome of the data as well as the original specimen. Particularly, the imaging of soft tissues with micro-CT might require tissue staining (<xref ref-type="bibr" rid="B63">Metscher 2009a</xref>, <xref ref-type="bibr" rid="B64">b</xref>), but neither the effects of contrast-enhancing chemicals nor of ionising radiation upon the integrity of tissue and genetic material are yet fully understood. Although micro-CT radiation seems to negatively affect the genetic material of living tissue (<xref ref-type="bibr" rid="B99">Wolff 1971</xref>, <xref ref-type="bibr" rid="B55">Kersemans et al. 2011</xref>), no fragmentation of the DNA could be detected in museum material of bird skins exposed to micro-CT scanning (<xref ref-type="bibr" rid="B73">Paredes et al. 2012</xref>). The morphological and molecular integrity of scanned material is particularly important when valuable museum material is imaged, otherwise the material is rendered useless for further investigations. Therefore, by testing whether treatment with contrast agents or exposure <pmc-comment>PageBreak</pmc-comment>to X-ray radiation create structural damage to the tissue of the sample or impair the potential to amplify nucleic acid structures important for the molecular identification, this study assesses the compliance of micro-CT imaging with the second requirement for creating a cybertype. Finally, various aspects of exploring and communicating the resulting information through new ways of publishing are demonstrated and evaluated with regard to the third requirement for a cybertype. The paper concludes by summarising both the potential and the shortcomings of micro-CT imaging for taxonomic research and provides an outlook to possible future developments, including the overall applicability of the cybertype concept and the establishment of virtual collections.</p></sec><sec sec-type="materials|methods"><title>Material and methods</title><sec sec-type="Specimen preparation and processing"><title>Specimen preparation and processing</title><p>Nine polychaete specimens (seven different species) were chosen for this study, all of them in the clade Aciculata (<named-content content-type="taxon-name">Annelida</named-content>, <named-content content-type="taxon-name">Polychaeta</named-content>) (<xref ref-type="table" rid="T1">Table 1</xref>). Specimens are stored in the collections of the biodiversity laboratory of the Hellenic Centre for Marine Research, except for those of <italic>Eunice roussaei</italic> Quatrefages, 1866 (deposited in the Aristotelian University of Thessaloniki), <italic><named-content content-type="taxon-name">Alitta succinea</named-content></italic> (Leuckart, 1847)and <italic><named-content content-type="taxon-name">Hermodice carunculata</named-content></italic> (Pallas, 1776) (both subsequently used for molecular analyses and destroyed). All specimens had originally been fixed and preserved in different media, but were dehydrated to 96% ethanol prior to treatment. Identification was performed to the lowest possible level under a stereo microscope and light microscope, using the most recent literature available for each taxon (e.g. <xref ref-type="bibr" rid="B15">Böggemann 2002</xref>, <xref ref-type="bibr" rid="B88">San Martín 2003</xref>, <xref ref-type="bibr" rid="B6">Bakken 2004</xref>, <xref ref-type="bibr" rid="B16">Carrera-Parra 2006a</xref>, <xref ref-type="bibr" rid="B100">Zanol and Bettoso 2006</xref>). However, no dissections were performed, in order to assess whether internal characters required for identification in several species could be determined through virtual dissections instead.</p><table-wrap id="T1" orientation="portrait" position="float"><label>Table 1.</label><caption><p>Overview of scanned polychaete specimens, their preparatory treatment and scanning parameters. † = maximum dimension (in any direction) of the scanned part of the specimen; ‡ = multi-part scan, automatically joined; § = distance between projection images in degrees; | = total size of cross-section images in megabyte, file format: PNG/BMP.</p></caption><table frame="hsides" rules="groups"><tbody><tr><td rowspan="1" colspan="1">Species</td><td rowspan="1" colspan="1">Sample code</td><td rowspan="1" colspan="1">Location and depth</td><td rowspan="1" colspan="1">Size (mm) <sup>†</sup></td><td rowspan="1" colspan="1">Scanned part</td><td rowspan="1" colspan="1">Sample preparation</td><td rowspan="1" colspan="1">Scanning medium</td><td rowspan="1" colspan="1">Rotation step <sup>§</sup></td><td rowspan="1" colspan="1">Number of projection images</td><td rowspan="1" colspan="1">Exposure time per image (ms)</td><td rowspan="1" colspan="1">Total scanning time (h:min)</td><td rowspan="1" colspan="1">Resolution (μm per pixel)</td><td rowspan="1" colspan="1">Dataset size <sup>|</sup></td></tr><tr><td rowspan="1" colspan="1"><italic><named-content content-type="taxon-name">Lumbrineris latreilli</named-content></italic> Audouin & Milne Edwards, 1834</td><td rowspan="1" colspan="1">ΦΗ20D</td><td rowspan="1" colspan="1">Gulf of Heraklion, Crete, Greece (<named-content content-type="dwc:verbatimCoordinates">35.3527°, 024.1084°</named-content>), 20m</td><td rowspan="1" colspan="1">2.4</td><td rowspan="1" colspan="1">Anterior end</td><td rowspan="1" colspan="1">none</td><td rowspan="1" colspan="1">air</td><td rowspan="1" colspan="1">0.25°</td><td rowspan="1" colspan="1">1440</td><td rowspan="1" colspan="1">1925</td><td rowspan="1" colspan="1">3:08</td><td rowspan="1" colspan="1">1.919</td><td rowspan="1" colspan="1">506(BMP)</td></tr><tr><td rowspan="1" colspan="1"><italic><named-content content-type="taxon-name">Eunice</named-content></italic> sp. (juvenile)</td><td rowspan="1" colspan="1">CALA-10b-08</td><td rowspan="1" colspan="1">Alykes, Crete, Greece (<named-content content-type="dwc:verbatimCoordinates">35.4158°, 024.9875°</named-content>), 10m</td><td rowspan="1" colspan="1">1.8</td><td rowspan="1" colspan="1">Whole animal</td><td rowspan="1" colspan="1">24 h in iodine</td><td rowspan="1" colspan="1">96% ethanol</td><td rowspan="1" colspan="1">0.26°</td><td rowspan="1" colspan="1">1384</td><td rowspan="1" colspan="1">1835 </td><td rowspan="1" colspan="1">3:33</td><td rowspan="1" colspan="1">2.162</td><td rowspan="1" colspan="1">69 (BMP)</td></tr><tr><td rowspan="1" colspan="1"><italic><named-content content-type="taxon-name">Eunice</named-content></italic> sp. (juvenile)</td><td rowspan="1" colspan="1">CELB-10c-08</td><td rowspan="1" colspan="1">Elounda, Crete, Greece (<named-content content-type="dwc:verbatimCoordinates">35.2516°, 025.7583°</named-content>), 1m</td><td rowspan="1" colspan="1">2.2</td><td rowspan="1" colspan="1">Whole animal</td><td rowspan="1" colspan="1">24 h in PTA</td><td rowspan="1" colspan="1">96% ethanol</td><td rowspan="1" colspan="1">0.3°</td><td rowspan="1" colspan="1">1200</td><td rowspan="1" colspan="1">1835</td><td rowspan="1" colspan="1">3:05</td><td rowspan="1" colspan="1">2.298</td><td rowspan="1" colspan="1">93(BMP)</td></tr><tr><td rowspan="1" colspan="1"><italic><named-content content-type="taxon-name">Eunice roussaei</named-content></italic> Quatrefages, 1866</td><td rowspan="1" colspan="1">POL/EUN/503</td><td rowspan="1" colspan="1">Thermaikos Gulf, Greece</td><td rowspan="1" colspan="1">2.9</td><td rowspan="1" colspan="1">parapodium (mid-body)</td><td rowspan="1" colspan="1">2 h in HDMS</td><td rowspan="1" colspan="1">air</td><td rowspan="1" colspan="1">0.25°</td><td rowspan="1" colspan="1">1440</td><td rowspan="1" colspan="1">1155</td><td rowspan="1" colspan="1">2:19</td><td rowspan="1" colspan="1">2.878</td><td rowspan="1" colspan="1">326(BMP)</td></tr><tr><td rowspan="1" colspan="1"><italic><named-content content-type="taxon-name">Alitta succinea</named-content></italic> (Leuckart, 1847)</td><td rowspan="1" colspan="1">F6BNR1</td><td rowspan="1" colspan="1">Etang de Berre, France (<named-content content-type="dwc:verbatimCoordinates">43.4699°, 005.1699°</named-content>), 2m </td><td rowspan="1" colspan="1">8.1</td><td rowspan="1" colspan="1">Anterior end</td><td rowspan="1" colspan="1">20 days in PTA, solution renewed every 5 days</td><td rowspan="1" colspan="1">96% ethanol</td><td rowspan="1" colspan="1">0.4°</td><td rowspan="1" colspan="1">900</td><td rowspan="1" colspan="1">190</td><td rowspan="1" colspan="1">2 x 0:17 <sup>‡</sup></td><td rowspan="1" colspan="1">3.425</td><td rowspan="1" colspan="1">1433(PNG)</td></tr><tr><td rowspan="1" colspan="1"><italic><named-content content-type="taxon-name">Phyllodoce lineata</named-content></italic> (Claparède, 1870)</td><td rowspan="1" colspan="1">Φ2Η25E</td><td rowspan="1" colspan="1">Gulf of Heraklion, Crete, Greece (<named-content content-type="dwc:verbatimCoordinates">35.3640°, 025.1086°</named-content>), 25m</td><td rowspan="1" colspan="1">3.9</td><td rowspan="1" colspan="1">Anterior end</td><td rowspan="1" colspan="1">36 h in PTA</td><td rowspan="1" colspan="1">96% ethanol</td><td rowspan="1" colspan="1">0.28°</td><td rowspan="1" colspan="1">1285</td><td rowspan="1" colspan="1">1165</td><td rowspan="1" colspan="1">2:05</td><td rowspan="1" colspan="1">1.645</td><td rowspan="1" colspan="1">450(PNG)</td></tr><tr><td rowspan="1" colspan="1"><italic><named-content content-type="taxon-name">Phyllodoce</named-content></italic> sp.</td><td rowspan="1" colspan="1">Φ2Η40D</td><td rowspan="1" colspan="1">Gulf of Heraklion, Crete, Greece (<named-content content-type="dwc:verbatimCoordinates">35.3832°, 025.1086°</named-content>), 40m</td><td rowspan="1" colspan="1">5.5</td><td rowspan="1" colspan="1">Anterior end and mid-body</td><td rowspan="1" colspan="1">36 h in PTA</td><td rowspan="1" colspan="1">96% ethanol</td><td rowspan="1" colspan="1">0.28°</td><td rowspan="1" colspan="1">1285</td><td rowspan="1" colspan="1">1336</td><td rowspan="1" colspan="1">2:04</td><td rowspan="1" colspan="1">1.096</td><td rowspan="1" colspan="1">2406(PNG)</td></tr><tr><td rowspan="1" colspan="1"><italic><named-content content-type="taxon-name">Syllis gracilis</named-content></italic> Grube, 1840</td><td rowspan="1" colspan="1">CELB-1a-07</td><td rowspan="1" colspan="1">Elounda, Crete, Greece (<named-content content-type="dwc:verbatimCoordinates">35.2516°, 025.7583°</named-content>), 1m</td><td rowspan="1" colspan="1">7.6</td><td rowspan="1" colspan="1">Anterior end and mid-body</td><td rowspan="1" colspan="1">3 days in iodine</td><td rowspan="1" colspan="1">96% ethanol</td><td rowspan="1" colspan="1">0.25°</td><td rowspan="1" colspan="1">1440</td><td rowspan="1" colspan="1">1165</td><td rowspan="1" colspan="1">3 x 1:52 <sup>‡</sup></td><td rowspan="1" colspan="1">1.439</td><td rowspan="1" colspan="1">2969(PNG)</td></tr><tr><td rowspan="1" colspan="1"><italic><named-content content-type="taxon-name">Hermodice carunculata</named-content></italic> (Pallas, 1776)</td><td rowspan="1" colspan="1">–</td><td rowspan="1" colspan="1">Alykes, Crete, Greece (<named-content content-type="dwc:verbatimCoordinates">35.4158°, 024.9875°</named-content>), 5m</td><td rowspan="1" colspan="1">10.1</td><td rowspan="1" colspan="1">Anterior end</td><td rowspan="1" colspan="1">2 x 2 h in HMDS</td><td rowspan="1" colspan="1">air</td><td rowspan="1" colspan="1">0.3°</td><td rowspan="1" colspan="1">1200</td><td rowspan="1" colspan="1">155</td><td rowspan="1" colspan="1">0:32</td><td rowspan="1" colspan="1">8.922</td><td rowspan="1" colspan="1">877 (BMP)</td></tr></tbody></table></table-wrap><p>To test the effect of different contrast-enhancement methods on the imaging results and tissue characteristics, several samples were treated with one of the following methods: a) tissue staining with 1% iodine in 96% ethanol; b) tissue staining with 0.3% phosphotungstic acid (PTA) in 70% ethanol; c) desiccation with Hexamethyldisilazane (HMDS). Protocols for both iodine and PTA staining follow <xref ref-type="bibr" rid="B63">Metscher (2009a)</xref>. In both solutions, smaller samples were stained in 2 ml for 24 hours to several days, larger samples in PTA required longer staining (up to 3 weeks) in larger amounts (10 ml), the solution was renewed every five days to allow PTA to penetrate into the tissue. Samples treated with HMDS were left in the chemical for two to four hours, in the larger specimen (<italic><named-content content-type="taxon-name">Hermodice carunculata</named-content></italic>) the chemical was renewed after two hours. The amount of HMDS and the treatment time depends on the size of the specimen: as a general guide, an amount twice the body volume of the specimen was used. Afterwards, specimens were removed from the chemical and left to dry for several hours, causing them to desiccate while retaining their morphology. Details on treatment for each specimen are presented in <xref ref-type="table" rid="T1">Table 1</xref>.</p><p><pmc-comment>PageBreak</pmc-comment><pmc-comment>PageBreak</pmc-comment>Wet samples were scanned in heat-sealed 200 µl polypropylene pipette tips, either in ethanol or in air. The top of the container was sealed with a plasticine cap to prevent the specimen from drying out during scanning (for a similar setup see <xref ref-type="bibr" rid="B63">Metscher 2009a</xref>). Samples dried with HMDS were partially enclosed in a small piece of styrofoam which in turn was mounted on a thin metallic sample holder. For assessing the quality of the scans with regard to distinguishing features, in this study only the anterior end of most worms was scanned. Scanning only the anterior end reduced scanning time and allowed us to choose a higher resolution. In polychaetes, the anterior end usually comprises most diagnostic characters, thus allowing us to assess the usefulness of the scans based on taxonomic criteria.</p></sec><sec sec-type="Image acquisition"><title>Image acquisition</title><p>Samples were imaged with a SkyScan 1172 microtomograph (<ext-link ext-link-type="uri" xlink:href="http://www.skyscan.be/products/1172.htm">http://www.skyscan.be/products/1172.htm</ext-link> ) at the biodiversity laboratory of the Hellenic Centre for Marine Research. This system uses a tungsten source with energies ranging from 20–100kV and is equipped with an 11 megapixel CCD camera (4000×2672 pixel) with a maximal resolution of <0.8 µm/pixel. Specimens were scanned at a voltage of 60 kV with a flux of 167µA and scans were performed for a full rotation of 360°. Except for <italic><named-content content-type="taxon-name">Alitta succinea</named-content></italic>, for which a camera pixel binning of 2 × 2 was chosen, images were always acquired at highest camera resolution. Individual scanning parameters can be found in <xref ref-type="table" rid="T1">Table 1</xref>. Projection images acquired during the scanning process were subsequently reconstructed into cross sections with SkyScan’s NRecon software which employs a modified Feldkamp’s back-projection algorithm. Sections were always reconstructed from the total number of projection images (360°) to obtain a greater level of detail, other reconstruction parameters were chosen individually for each sample. In case of strong density differences in the scanned sample, the upper limit of the grey scale histogram was lowered to unite very dense values. This causes dense values above the set limit to be assigned to the same grey scale value without differentiation and allows softer (less dense) tissues to be visualised with greater detail. The lower limit of the histogram was set at the value for the surrounding medium (air or ethanol). To reduce the size of the resulting images, only areas containing relevant data (regions of interest) were reconstructed, thus excluding the surrounding air or enclosure medium.</p></sec><sec sec-type="Molecular analyses"><title>Molecular analyses</title><p><italic><named-content content-type="taxon-name">Hediste diversicolor</named-content></italic> (O.F. Müller, 1776) specimens collected in Tsopeli lagoon in Amvrakikos Gulf (Western Greece) were sequenced before and after X-ray exposure in order to assess whether the radiation had an effect on the 16S rRNA sequence obtained. Samples were exposed either to high energy of radiation for a relatively short time (100kV for 1.5h) or repetitively exposed for three cycles of 12 hours at medium <pmc-comment>PageBreak</pmc-comment>energy (12h, 24h, 36h at 60kV). In the latter series, some tissue was removed from the specimen for DNA extraction after each cycle. A fragment of the 16S rRNA gene (~ 500 bp) was amplified using a primer pair designed for polychaetes: 16SAN-F (TACCTTTTCATCATGG) and 16SEU-R (ACCTTTGCACGGTCAGGRTACCGC) (<xref ref-type="bibr" rid="B101">Zanol et al. 2010</xref>). Genomic DNA concentration of samples was calculated using a Nanodrop 1000 spectrophotometer (Wilmington DE, USA) on wavelength 260 and 280nm. Polymerase Chain Reactions (PCRs) were performed in a final volume of 20 µl containing 0.4 units KAPA Taq DNA polymerase (Kapa Biosystems, Inc, USA), 2 µl PCR buffer 10x, 4.5 mM MgCl2, 2 mM dNTPs, 10 pmol of each primer, and 0.5 µl (~5 ng/μl) of DNA template. Amplification was performed in a MJ Research PTC-200 Thermo Cycler (Harlow Scientific, USA) programmed as follows: initial denaturation cycle at 96°C (4 min) followed by 35 cycles of denaturation at 93°C (45 sec), annealing at 52°C (1 min) and extension at 72°C (1 min); last cycle was followed by a final extension at 72°C for 7 min. The sequences were processed with MEGA v. 5 software (<xref ref-type="bibr" rid="B92">Tamura et al. 2011</xref>). Obtained sequences were submitted to GenBank (<xref ref-type="bibr" rid="B9">Benson et al. 2005</xref>) under the accession numbers KC113440-KC113445.</p></sec><sec sec-type="Processing, presentation and dissemination of image data"><title>Processing, presentation and dissemination of image data</title><p><bold>Two-dimensional images:</bold></p><p>All resulting datasets of cross sections were post-processed with the CTAnalyzer (CTAn) software (SkyScan, Kontich, Belgium) by selecting a Region of Interest (ROI) containing the sample but removing further superfluous information, thus creating a dataset of reduced size. To obtain a three-dimensional representation of the sequence of cross section images, the data were visualised with two different volume rendering software packages: both CTVox (SkyScan, Kontich, Belgium) and the free software Drishti were employed (<ext-link ext-link-type="uri" xlink:href="http://anusf.anu.edu.au/Vizlab/drishti/">http://anusf.anu.edu.au/Vizlab/drishti/</ext-link> ). Volume rendering displays the data by assigning a colour value and an opacity value to each data point (voxel) in the dataset. By changing these transfer values, different features of the dataset can be visualised and explored. Density-based false-colour renderings were applied to the data where this was considered helpful to visualise structures. Isosurface models (geometrical representations of surfaces of equal values) were created with Amira v. 5.2 (Visage Imaging, Berlin, Germany). Two-dimensional images were extracted as bitmap files with the image export function of the respective software and consequently cropped to final dimensions and minimally edited in Adobe Photoshop to enhance contrast (adjusting image levels and curves) or transform colour tint (adjusting hue and saturation), as well as to add annotations.</p><p><bold>Interactive volumetric data:</bold></p><p>The <italic><named-content content-type="taxon-name">Lumbrineris latreilli</named-content></italic> dataset was first processed with custom functions of CTAn (thresholding, smoothing, noise removal) to isolate the jaws from the surrounding tissue and saved as a separate dataset. This new dataset was subsequently loaded into <pmc-comment>PageBreak</pmc-comment>the free image editor Fiji (<ext-link ext-link-type="uri" xlink:href="http://fiji.sc">http://fiji.sc</ext-link> ) and reduced in size to a stack of 320 images with dimensions of 205 × 173 pixels. These bitmaps were converted into TGA (Truevision Graphics Adapter) files with the free ImageMagick tool <ext-link ext-link-type="uri" xlink:href="http://www.imagemagick.org">http://www.imagemagick.org</ext-link> ) and rendered with the C++ programmevolren (<xref ref-type="bibr" rid="B86">Ruthensteiner et al. 2010</xref>) which is based on the plotting library S2PLOT (<xref ref-type="bibr" rid="B7">Barnes et al. 2006</xref>). With this library, the data were converted into a three-dimensional VRML (Virtual Reality Modelling Language) object, accompanied by a PNG (Portable Network Graphics) file of each angular view. A corresponding script provided by the authors ensures that the correct view is rendered when the object is manipulated. In the volren script parameters were adjusted to AMIN=0.0001 and AMAX=0.1 and the colour map “iron” was chosen.In the resulting VRML file the texture transparency parameter was changed from 0.4 to 0.1 throughout the file in order to increase the contrast of the embedded model in the PDF (Portable Document Format) file.</p><p><bold>Interactive surface description models:</bold></p><p>Using the segmentation editor of Amira, features of interest were manually segmented (“labelled”) with the brush tool. For each feature a new LabelField was created, thus allowing the different objects to be manipulated separately at later stages. Labelled features were converted into surfaces with the SurfaceGen module and where the number of polygons was too high (>1,000,000) they were reduced with the Simplifier tool to increase computation efficiency during further processing. Amira’s SmoothSurface module did not produce satisfactory results in models with small detailed structures, since the module does not allow for selective smoothing and small structures disappeared after the application of the module. The models were, therefore, exported as OBJ (Wavefront Object) files and further processed with Blender 2.63a (<ext-link ext-link-type="uri" xlink:href="http://www.blender.org">http://www.blender.org</ext-link> ), a high end, open source, 3D design program. In Blender, the surface was cleaned of artefacts by applying the Vertices’ Relaxation and the Vertices Smoothing operations, which replaced the original model’s points (vertices) in average positions between them, thus smoothing surface anomalies. This process was repeated where necessary, until the model’s surface appeared smooth and even, without alienating main parts or the overall morphology of the model. If surface noise still persisted in parts of the model, then a second, manual part of cleaning was applied. In this case, specific parts of the model were selected individually and corrected by using additional tools in Blender (e.g. sculpting smooth brush). Some geometrically elegant (small, narrow, light) parts of the model (e.g. chaetae) could not be cleaned or were destroyed by the above techniques, in this case these parts and elements were reconstructed (remodelled) manually and added again to the model.</p><p><bold>Videos:</bold></p><p>The videos of 3D volume renderings were created with CTVox, using the flight recorder function, and saved as an AVI (Audio Video Interface) file. The video of cross sections through the sample was created by loading the image stack of the sections into Fiji and re-sampling the data to reduce image size. The resulting new stack was exported as an AVI video file with 20 frames per second. To be able to embed the vid<pmc-comment>PageBreak</pmc-comment>eos into the PDF document, the AVI files were converted with an online conversion software (<ext-link ext-link-type="uri" xlink:href="http://www.online-convert.com">http://www.online-convert.com</ext-link> ) into FLV (Flash Video) files with a bit rate of 1000kb/s and a width of 400px.</p><p><bold>Embedding multimedia and interactive objects into PDFs:</bold></p><p>Both surface models and volume renderings were embedded into the PDF with the Acrobat X Pro 3D PDF Converter Suite (Tetra 4D, Seattle, USA). The 3D Reviewer module was used to define colours and views and to add annotations. The resulting data were again exported to PDF format, specifying in the export options “PCR tessellation” and “Compress tessellation”. These options reduced the final file size to about one third of the original object size. The proper rendering of the volumetric data required also the inclusion of a JavaScript file which is distributed along with the S2PLOT library (<ext-link ext-link-type="uri" xlink:href="http://astronomy.swin.edu.au/s2plot/peripheral/s2plot.js">http://astronomy.swin.edu.au/s2plot/peripheral/s2plot.js</ext-link> ). Videos were added with Adobe’s Add Multimedia function. The process of embedding models and multimedia content into PDFs is relatively straightforward, detailed descriptions are provided by <xref ref-type="bibr" rid="B7">Barnes et al. (2006)</xref>, <xref ref-type="bibr" rid="B87">Ruthensteiner and Hess (2008)</xref>, <xref ref-type="bibr" rid="B57">Kumar et al. (2010)</xref> and <xref ref-type="bibr" rid="B86">Ruthensteiner et al. (2010)</xref>.</p><p><bold>Electronic publication and data dissemination:</bold></p><p>All media included in this publication as well as supporting material (surface models, image files, videos) are published under a Creative Commons Attribution 3.0 (CC-BY) licence in a Virtual Research Environment, the Polychaete Scratchpad (<ext-link ext-link-type="uri" xlink:href="http://polychaetes.marbigen.org">http://polychaetes.marbigen.org</ext-link> ). The full volumetric datasets have been archived at the Dryad Data Repository (<ext-link ext-link-type="uri" xlink:href="http://datadryad.org">http://datadryad.org</ext-link> , doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.5061/dryad.84m54">10.5061/dryad.84m54</ext-link>). Since the direct inclusion of interactive, three-dimensional models in web pages is still in its infancy and requires specific browser and driver configurations on the client side, interactive models have been included as separate PDF files on the web site of the journal as well as on the Scratchpad site, thus allowing the majority of users to access this content. Most Acrobat products (Reader, Professional) from Version 8 onwards support the display of embedded media. However, specific versions of the software still show incompatibility problems and some users might therefore encounter problems viewing the interactive content. In this case, it is recommended to download the multimedia content (videos, interactive models) from Dryad, or the Polychaete Scratchpad, and view it with other software (e.g. a multimedia player).</p></sec></sec><sec sec-type="Results"><title>Results</title><sec sec-type="Information content of the datasets"><title>Information content of the datasets</title><p><bold><italic><named-content content-type="taxon-name">Lumbrineris latreilli</named-content></italic>:</bold></p><p>Only the anterior end of the specimen was scanned, the total length of the visualised part being 2.4 mm. At a resolution of 1.9 µm/pixel, the smallest discernible structures <pmc-comment>PageBreak</pmc-comment>are about 4 µm in size. Since the specimen has been scanned in air without prior desiccation, a thin film of ethanol partially covers the body and obscures parts of the external morphology; however, general external characters (size and shape of larger features such as segments, head, mouth opening) are clearly recognisable (<xref ref-type="fig" rid="F1">Fig. 1a</xref>). The shape of the parapodia is visible but obscured by the ethanol film. The number and arrangement of the chaetae can be discerned but the chosen resolution prevents finer details such as chaetal articulation or dentation from being captured. The internal anatomy, on the contrary, has been recorded in substantial detail. The resolution is high enough to allow the observation of even the fine structure of the vascular system in the anterior part of the head (<xref ref-type="fig" rid="F1">Figs 1b–c</xref>). Likewise, muscular groups and even their fibres are clearly visible (<xref ref-type="fig" rid="F1">Fig. 1d</xref>). Nervous tissues (brain, ganglia, nerves) have a very low density and are difficult to depict in detail in the volume renderings. However, large nervous <pmc-comment>PageBreak</pmc-comment>structures can be visualised by applying appropriate transfer functions (<xref ref-type="fig" rid="F2">Video 1</xref>) and through careful remodelling (<xref ref-type="fig" rid="F1">Fig. 1b</xref>). The most prominent visible feature is the jaw apparatus, a calcified complex system of maxillae and mandibles (<xref ref-type="fig" rid="F3">Figs 2</xref><xref ref-type="fig" rid="F4"></xref><xref ref-type="fig" rid="F5">–4</xref>). However, the maxillary pair MV, the maxillary carriers, as well as the accessory lamellae of mandibles III and IV appear to have a similar density to the surrounding muscle tissue. This makes it impossible to visualise them in the low-resolution interactive volume model included in this publication (<xref ref-type="fig" rid="F4">Fig. 3</xref>). In the high-resolution dataset, however, they can be visualised by applying suitable transfer functions (<xref ref-type="fig" rid="F5">Figs 4b–c</xref>), which allows them to be included in the surface model (<xref ref-type="fig" rid="F3">Fig. 2</xref>). The accessory lamellae of MI and the connecting plates of MI and MII, described as “weakly sclerotised in <italic><named-content content-type="taxon-name">Lumbrineris</named-content></italic>” by <xref ref-type="bibr" rid="B17">Carrera-Parra (2006b)</xref>, cannot not be clearly discerned in the data.</p><fig id="F1" orientation="portrait" position="float"><label>Figure 1.</label><caption><p><italic><named-content content-type="taxon-name">Lumbrineris latreilli</named-content></italic>, <bold>a</bold> false-colour volume rendering of the anterior region, dorso-lateral view, specimen partly covered by a thin film of ethanol <bold>b</bold> surface model of the vascular system (yellow) and brain (purple) superimposed on volume rendering of the worm, dorso-anterior view <bold>c</bold> false-colour, semi-transparent volume rendering of the anterior region, dorsal view <bold>d</bold> volume rendering of virtually dissected anterior region, lateral view.</p></caption><graphic xlink:href="ZooKeys-263-001-g001"></graphic></fig><fig id="F2" orientation="portrait" position="float"><label>Video 1.</label><caption><p><italic><named-content content-type="taxon-name">Lumbrineris latreilli</named-content></italic>, false-colour volume rendering, virtual dissection of anterior end, dorsal view. Video available for download in full resolution from <ext-link ext-link-type="uri" xlink:href="http://polychaetes.marbigen.org/lumbrineris-latreilli-micro-ct-video">http://polychaetes.marbigen.org/lumbrineris-latreilli-micro-ct-video</ext-link> .</p></caption><media xlink:href="ZooKeys-263-001-g002.flv" orientation="portrait" xlink:type="simple" id="d35e1033" position="anchor" mimetype="video" mime-subtype="x-flv"></media></fig><fig id="F3" orientation="portrait" position="float"><label>Figure 2.</label><caption><p><italic><named-content content-type="taxon-name">Lumbrineris latreilli</named-content></italic>, surface model of the jaw apparatus. Terminology follows Carrera-Parra (2006). If viewed with Adobe Acrobat Reader (version 8 or higher), the interactive 3D-mode can be activated by clicking on the image, allowing the user to rotate, move and magnify the model, to isolate elements and to change the light settings.</p></caption><media xlink:href="ZooKeys-263-001-g003.pdf" orientation="portrait" xlink:type="simple" id="d35e1043" position="anchor" mimetype="application" mime-subtype="pdf"></media></fig><fig id="F4" orientation="portrait" position="float"><label>Figure 3.</label><caption><p><italic><named-content content-type="taxon-name">Lumbrineris latreilli</named-content></italic>, volume model of the jaw apparatus. Note the absence of MV as well as the accessory lamellae of MI, MIII and MIV which cannot be displayed in this low-resolution version. If viewed with Adobe Acrobat Reader (version 8 or higher), the interactive 3D-mode can be activated by clicking on the image, allowing the user to rotate, move and magnify the model.</p></caption><media xlink:href="ZooKeys-263-001-g004.pdf" orientation="portrait" xlink:type="simple" id="d35e1053" position="anchor" mimetype="application" mime-subtype="pdf"></media></fig><fig id="F5" orientation="portrait" position="float"><label>Figure 4.</label><caption><p><italic><named-content content-type="taxon-name">Lumbrineris latreilli</named-content></italic>, volume rendering of <bold>a</bold> anterior regionwith maxillary apparatus superimposed as a surface model, dorsal view <bold>b</bold> virtually dissected jaw apparatus, ventral view with mandibles partly removed <bold>c</bold> same, dorsal view.</p></caption><graphic xlink:href="ZooKeys-263-001-g005"></graphic></fig><p><bold><italic><named-content content-type="taxon-name">Eunice</named-content></italic> sp. (juveniles):</bold></p><p>The specimens were similar in size and developmental stage, making them ideal for testing the effect of different tissue stains (discussed below). As in the <italic><named-content content-type="taxon-name">Lumbrineris</named-content></italic> dataset, larger external features such as body shape, antennae and parapodia are excel<pmc-comment>PageBreak</pmc-comment>lently visible (<xref ref-type="fig" rid="F6">Figs 5a–b</xref>), although the animals are smaller and the resolution is slightly coarser (ca 2.25 µm/pixel). No details of chaetae are visible, but the subacicular hooks are clearly depicted especially in the iodine-stained specimen (<xref ref-type="fig" rid="F6">Fig. 5b</xref>) and the chaetal bundle is well visible in cross sections (<xref ref-type="fig" rid="F7">Fig. 6</xref>). Internally, large organs and muscle groups are fairly distinct (<xref ref-type="fig" rid="F7">Fig. 6</xref>), but the small size of the animal (width ca 0.3 mm) does not allow details that would be visible in histological sections to be discerned (e.g. vascular system, ganglia), with the exception of muscle fibres. The general structure of the jaw apparatus is evident, but finer details are difficult to see, and only MI–MIII can be unambiguously identified (<xref ref-type="fig" rid="F6">Fig. 5c</xref>).</p><fig id="F6" orientation="portrait" position="float"><label>Figure 5.</label><caption><p><italic><named-content content-type="taxon-name">Eunice</named-content></italic> sp., false-colour volume renderings (colours indicate relative densities: blue: low, green: medium, red: high) of <bold>a</bold> iodine-stained specimen, ventral view <bold>b</bold> PTA-stained specimen, ventral view <bold>c</bold> false-colour volume rendering of virtually dissected maxillary apparatus (iodine-stained individual), dorso-lateral view.</p></caption><graphic xlink:href="ZooKeys-263-001-g006"></graphic></fig><fig id="F7" orientation="portrait" position="float"><label>Figure 6.</label><caption><p><italic><named-content content-type="taxon-name">Eunice</named-content></italic> sp., transaxial cross section at mid-body (iodine-stained individual).</p></caption><graphic xlink:href="ZooKeys-263-001-g007"></graphic></fig><p><bold><italic><named-content content-type="taxon-name">Eunice roussaei</named-content></italic>:</bold></p><p>The dissected parapodium shows, especially in the cross sections, the exact arrangement of the different chaetal types (aciculae, subacicular hook, compound chaetae, supra-ac<pmc-comment>PageBreak</pmc-comment><pmc-comment>PageBreak</pmc-comment>icular limbate and pectinate chaetae as well as internal notopodial chaetae) (<xref ref-type="fig" rid="F8">Figs 7a–b</xref>). External features, such as dorsal and ventral cirri, parapodial lobes, branchial stem and branchial filaments, can likewise be observed (<xref ref-type="fig" rid="F8">Fig. 7c</xref>). Due to the position in which the specimen dried, the branchial stem is slightly recoiled, making the exact count of <pmc-comment>PageBreak</pmc-comment>branchial filaments not straightforward; they are better delineated in the interactive, three-dimensional model of the specimen (<xref ref-type="fig" rid="F9">Fig. 8</xref>). Details of chaetae (e.g. serration, dentation) are not visible (<xref ref-type="fig" rid="F8">Fig. 7d</xref>). This is primarily caused by the coarse resolution of the scan (ca 2.9 μm/pixel), which prevents capture of these tiny details. Furthermore, because the specimen was dried and scanned in air, the chaetae vibrate slightly during each rotation step, resulting in a slight blur in the final images (<xref ref-type="fig" rid="F8">Fig. 7d</xref>). Several internal structures are visible, such as muscle groups or connective tissues, but no blood vessels (e.g. in the branchiae) can be seen. Generally, most internal structures are difficult to identify, since the contrast between neighbouring tissues was equalised by the desiccation process.</p><fig id="F8" orientation="portrait" position="float"><label>Figure 7.</label><caption><p><italic><named-content content-type="taxon-name">Eunice roussaei</named-content></italic>, mid-body parapodium, <bold>a</bold> cross section through parapodial base <bold>b</bold> semi-transparent volume rendering <bold>c</bold> opaque volume rendering <bold>d</bold> volume rendering of chaetae.</p></caption><graphic xlink:href="ZooKeys-263-001-g008"></graphic></fig><fig id="F9" orientation="portrait" position="float"><label>Figure 8.</label><caption><p><italic><named-content content-type="taxon-name">Eunice roussaei</named-content></italic>, surface model of mid-body parapodium. Limbate chaetae, pectinate chaetae and notochaetae are not shown. Compound chaetae have been included as simplified, remodelled version and do not depict true shapes. If viewed with Adobe Acrobat Reader (version 8 or higher), the interactive 3D-mode can be activated by clicking on the image, allowing the user to rotate, move and magnify the model, to isolate elements or to change the light settings.</p></caption><media xlink:href="ZooKeys-263-001-g009.pdf" orientation="portrait" xlink:type="simple" id="d35e1193" position="anchor" mimetype="application" mime-subtype="pdf"></media></fig><p><bold><italic><named-content content-type="taxon-name">Alitta succinea</named-content></italic>:</bold></p><p>The overall morphology of the specimen, especially cephalic features (shape, appendages) and parapodia, can be clearly observed. The true three-dimensional structure of <pmc-comment>PageBreak</pmc-comment>the parapodia, which consist of a complex arrangement of parapodial lobes, as well as dorsal and ventral cirri, are best distinguished in stereo display (e.g. with anaglyph 3D glasses) (<xref ref-type="fig" rid="F10">Figs 9a–b</xref>, visible with red-cyan glasses). Chaetal structures cannot be observed, specimen preparation as well as scanning parameters were inappropriate for their proper visualisation. The pharynx in this specimen is everted, thus the determination of the shape of the jaws as well as of the paragnath shapes (conical) and distribution patterns—important taxonomic characters—is straightforward without having to virtually dissect the specimen (<xref ref-type="fig" rid="F10">Fig. 9a</xref>). Since X-ray imaging is attenuation-based and thus records differences in density (or, depending on the energy, differences in the atomic number of the material), the colour of the paragnaths (another diagnostic character) cannot be observed. Eyes are visible as slightly darker spots on the epidermis, their density is apparently slightly different from the surrounding tissue (<xref ref-type="fig" rid="F10">Fig. 9d</xref>). The internal anatomy is, as in the other samples, well visible. Major muscle groups can be discerned (<xref ref-type="fig" rid="F10">Fig. 9c, e</xref>), as well as the vascular system. The latter seems to be discontinuous in parts (<xref ref-type="fig" rid="F10">Fig. 9c</xref>), possibly a result of fixation in ethanol. The brain or other nervous tissues cannot be discerned. Apparently hollow regions in the pharyngeal area result from insufficient staining (<xref ref-type="fig" rid="F10">Fig. 9f</xref>). The density of these unstained regions is too low to be visualised and the corresponding information is lost.</p><fig id="F10" orientation="portrait" position="float"><label>Figure 9.</label><caption><p><italic><named-content content-type="taxon-name">Alitta succinea</named-content></italic>, <bold>a</bold> normal view and <bold>b</bold> stereo view of volume-rendering of anterior region, lateral view (3D effect is revealed when viewed with red-cyan glasses) <bold>c</bold> coronal cross section of anterior region depicting areas of insufficient tissue staining with PTA. Black areas in pharyngeal region remained unstained and information is lost <bold>d</bold> dorsal view and <bold>e</bold> posterior-lateral view of false-colour volume rendering of virtually dissected anterior region <bold>f</bold> volume rendering of anterior region, dorsal view.</p></caption><graphic xlink:href="ZooKeys-263-001-g010"></graphic></fig><pmc-comment>PageBreak</pmc-comment>
<pmc-comment>PageBreak</pmc-comment>
<p><bold><italic><named-content content-type="taxon-name">Phyllodoce lineata</named-content></italic> and <italic><named-content content-type="taxon-name">Phyllodoce</named-content></italic> sp.:</bold></p><p><italic><named-content content-type="taxon-name">Phyllodoce lineata</named-content></italic> can be identified to species level almost solely on the basis of the virtual specimen; all taxonomic key characters are visible, with the exception of mid-body parapodia which were not included in the imaged part of the specimen. However, the number, shape, length and arrangement of tentacular cirri and antennae, the shape (fused, partly fused, covered) of the prostomium and the anterior segments of the parapodia with dorsal and ventral cirri, and the position of chaetae can be observed (<xref ref-type="fig" rid="F12">Fig. 10a</xref>), but finer details of the parapodial lobes and the chaetae cannot. The pharynx is partly everted, which makes the assessment of the arrangement of the pharyngeal papillae straightforward. The subdivision of the pharynx is well visible, with the proximal end being covered with scattered smaller papillae, the distal end with six rows of large papillae (<xref ref-type="fig" rid="F12">Fig. 10a</xref>). The papillae around the pharyngeal opening can only be seen when the animal is virtually dissected (<xref ref-type="fig" rid="F12">Fig. 10b</xref>). The eyes, containing lenses as in most phyllodocid species (Rouse and Pleijel 2001), appear as dense structures (<xref ref-type="fig" rid="F12">Figs 10c–e</xref>, <xref ref-type="fig" rid="F11">Video 2</xref>). Internally, all muscular features, the gastrointestinal tract, ganglia and the <pmc-comment>PageBreak</pmc-comment><pmc-comment>PageBreak</pmc-comment>brain and large connecting nerves are well visible (<xref ref-type="fig" rid="F12">Figs 10d–e</xref>, <xref ref-type="fig" rid="F13">11a</xref>, <xref ref-type="fig" rid="F11">Video 2</xref>), especially when viewed in stereo display with red-cyan glasses (<xref ref-type="fig" rid="F12">Fig. 10e</xref>). The other <italic><named-content content-type="taxon-name">Phyllodoce</named-content></italic> specimen was imaged with a similar resolution and can thus be compared to the scan of <italic><named-content content-type="taxon-name">Phyllodoce lineata</named-content></italic>. Morphologically, no differences between the two specimens are evident, however, the pharynx in <italic><named-content content-type="taxon-name">Phyllodoce</named-content></italic> sp. is not everted. Virtual dissections reveal two subdivisions of the pharynx (<xref ref-type="fig" rid="F13">Fig. 11b</xref>), a distal part covered with large papillae and a proximal part with small papillae. The cross sections of the pharyngeal regions show that the distal part has six rows of large papillae (<xref ref-type="fig" rid="F13">Fig. 11c</xref>), however, neither the number nor arrangement of the smaller papillae can be determined (<xref ref-type="fig" rid="F13">Figs 11d</xref>), making an unambiguous identification of the species impossible. Naturally, no colour patterns — a species-specific character in several phyllodocid taxa (<xref ref-type="bibr" rid="B69">Nygren et al. 2010</xref>, <xref ref-type="bibr" rid="B70">Nygren and Pleijel 2011</xref>) — are visible in any of the scans.</p><fig id="F11" orientation="portrait" position="float"><label>Video 2.</label><caption><p><italic><named-content content-type="taxon-name">Phyllodoce lineata</named-content></italic>, false-colour volume rendering, virtual dissection. Video available for download in full resolution from <ext-link ext-link-type="uri" xlink:href="http://polychaetes.marbigen.org/phyllodoce-lineata-micro-ct-video">http://polychaetes.marbigen.org/phyllodoce-lineata-micro-ct-video</ext-link> .</p></caption><media xlink:href="ZooKeys-263-001-g011.flv" orientation="portrait" xlink:type="simple" id="d35e1341" position="anchor" mimetype="video" mime-subtype="x-flv"></media></fig><fig id="F12" orientation="portrait" position="float"><label>Figure 10.</label><caption><p><italic><named-content content-type="taxon-name">Phyllodoce lineata</named-content></italic>, <bold>a</bold> false-colour volume rendering of anterior region, dorso-lateral view<bold>b</bold> false-colour volume rendering of virtually dissected distal end of partly everted pharynx, focusing on terminal pharyngeal papillae, anterior view <bold>c</bold> transaxial cross section at eye level <bold>d</bold> volume rendering of virtually dissected anterior region showing the brain, dorso-posterior view <bold>e</bold> same, in stereo view (3D effect is revealed when viewed with red-cyan glasses).</p></caption><graphic xlink:href="ZooKeys-263-001-g012"></graphic></fig><fig id="F13" orientation="portrait" position="float"><label>Figure 11.</label><caption><p><bold>a</bold><italic><named-content content-type="taxon-name">Phyllodoce lineata</named-content></italic>, volume rendering of <bold>a</bold> anterior end, virtually dissected behind prostomial region, posterior view <bold>b</bold><italic><named-content content-type="taxon-name">Phyllodoce</named-content></italic> sp., volume rendering of virtually dissected pharynx, dorsal view <bold>c</bold><italic><named-content content-type="taxon-name">Phyllodoce</named-content></italic> sp., virtual dissection of distal pharyngeal subdivision with large papillae, anterior view <bold>d</bold><italic><named-content content-type="taxon-name">Phyllodoce</named-content></italic> sp., virtual dissection of proximal pharyngeal subdivision with small papillae, antero-lateral view.</p></caption><graphic xlink:href="ZooKeys-263-001-g013"></graphic></fig><p><bold><italic><named-content content-type="taxon-name">Syllis gracilis</named-content></italic>:</bold></p><p>As in the other data, large external morphological features of the specimen are well defined. Appendages, their articulation and number of articles are clearly depicted (<xref ref-type="fig" rid="F14">Fig. 12a</xref>). Internally, features such as muscles (<xref ref-type="fig" rid="F14">Figs 12b, d</xref>), the brain (<xref ref-type="fig" rid="F14">Fig. 12c</xref>), the gastrointestinal tract (<xref ref-type="fig" rid="F14">Fig. 12d</xref>) and muscle groups with their individual fibres can be identified. Other features such as eyes, pharyngeal papillae and the pharyngeal tooth are difficult to detect in volume renderings but can be observed in cross sections (<xref ref-type="fig" rid="F15">Figs 13a–c</xref>). <pmc-comment>PageBreak</pmc-comment>The length of the proventricle and the number of its muscle rows are likewise important diagnostic systematic characters in the family. The strong muscle fibres of the proventricle are extremely well visible, and through virtual dissection their three-dimensional <pmc-comment>PageBreak</pmc-comment>arrangement as well as their number can be well observed (<xref ref-type="fig" rid="F16">Fig. 14</xref>). However, one of the key characters in syllid systematics are fine differences in chaetal structures. These cannot be discerned with the present resolution; only the rather large and robust Y-shaped chaetae typical for <italic><named-content content-type="taxon-name">Syllis gracilis</named-content></italic> are visible (<xref ref-type="fig" rid="F14">Fig. 12e</xref>).</p><fig id="F14" orientation="portrait" position="float"><label>Figure 12.</label><caption><p><italic><named-content content-type="taxon-name">Syllis gracilis</named-content></italic>, false-colour volume rendering of <bold>a</bold> anterior region, anterior view <bold>b</bold> anterior region virtually dissected at pharyngeal level, posterior view <bold>c</bold> virtually dissected anterior region showing the brain, anterior view <bold>d</bold> virtually dissected region in front of proventricle, dorsal view; e) mid-body parapodia, posterior-lateral view.</p></caption><graphic xlink:href="ZooKeys-263-001-g014"></graphic></fig><fig id="F15" orientation="portrait" position="float"><label>Figure 13.</label><caption><p><italic><named-content content-type="taxon-name">Syllis gracilis</named-content></italic>, cross sections, <bold>a</bold> coronal view showing prostomium, palps and posterior eye pair <bold>b</bold> transaxial view at level of pharyngeal opening, showing papillae around pharyngeal opening <bold>c</bold> dorsal view, pharyngeal opening and pharyngeal tooth. Double lines at borders of cirri are artefacts resulting from either movement of specimen during the scan or from settings during dataset reconstruction.</p></caption><graphic xlink:href="ZooKeys-263-001-g015"></graphic></fig><fig id="F16" orientation="portrait" position="float"><label>Figure 14.</label><caption><p><italic><named-content content-type="taxon-name">Syllis gracilis</named-content></italic>, false-colour volume rendering of virtually dissected proventricle.</p></caption><graphic xlink:href="ZooKeys-263-001-g016"></graphic></fig><p><bold><italic><named-content content-type="taxon-name">Hermodice carunculata</named-content></italic>:</bold></p><p>This specimen has been scanned at a low resolution (8.9 µm/pixel) because of its large size, thus details of smaller structures such as chaetae are lost. Individual chaetae can be differentiated, but at the base of the parapodia they appear as a thick, merged structure (<xref ref-type="fig" rid="F17">Figs 15a, d</xref>). Larger external morphological features such as branchiae, parapodial structures, antennae and the caruncle are all clearly visible (<xref ref-type="fig" rid="F17">Fig. 15a</xref>). Eyes, <pmc-comment>PageBreak</pmc-comment>located under the cuticle, are visible in virtual dissection (<xref ref-type="fig" rid="F17">Fig. 15b</xref>). Internally, the heavily vascularised area around the pharynx is well defined (<xref ref-type="fig" rid="F17">Fig. 15c</xref>, <xref ref-type="fig" rid="F18">Video 3</xref>). The complex folds of the pharyngeal system can be observed (<xref ref-type="fig" rid="F17">Fig. 15b</xref>, <xref ref-type="fig" rid="F18">Videos 3</xref><xref ref-type="fig" rid="F19">–4</xref>), as well as various parts of the muscular system (pharyngeal muscles, longitudinal muscles, parapodial muscles). The brain is clearly visible (<xref ref-type="fig" rid="F17">Fig. 15b</xref>) and large nervous fibres such as the circumesophageal connective can be traced from the brain to the ventral ganglia (<xref ref-type="fig" rid="F17">Fig. 15d</xref>).</p><fig id="F17" orientation="portrait" position="float"><label>Figure 15.</label><caption><p><italic><named-content content-type="taxon-name">Hermodice carunculata</named-content></italic>, false-colour volume rendering of <bold>a</bold> anterior region, antero-dorsal view <bold>b</bold> virtually dissected anterior end showing pharyngeal structures, lateral view <bold>c</bold> virtually dissected anterior end, lateral view <bold>d</bold> virtually dissected anterior end at level of mouth opening, anterior view.</p></caption><graphic xlink:href="ZooKeys-263-001-g017"></graphic></fig><fig id="F18" orientation="portrait" position="float"><label>Video 3.</label><caption><p><italic><named-content content-type="taxon-name">Hermodice carunculata</named-content></italic>, false-colour volume rendering, virtual dissection of anterior end, dorsal view. Video available for download in full resolution from <ext-link ext-link-type="uri" xlink:href="http://polychaetes.marbigen.org/hermodice-carunculata-micro-ct-video">http://polychaetes.marbigen.org/hermodice-carunculata-micro-ct-video</ext-link> .</p></caption><media xlink:href="ZooKeys-263-001-g018.flv" orientation="portrait" xlink:type="simple" id="d35e1569" position="anchor" mimetype="video" mime-subtype="x-flv"></media></fig></sec><sec sec-type="Contrast enhancement"><title>Contrast enhancement</title><p>Two different contrast enhancement techniques were employed in this study. These involved the removal of the surrounding liquid medium and tissue staining with electron-dense substances. Both result in an increased density difference between the specimen and the surrounding medium and thus produce sharp, contrasting images. However, different methods accentuate different morphological and anatomical features and thus the information content of the data differs accordingly. <italic><named-content content-type="taxon-name">Lumbrineris latreilli</named-content></italic> was simply scanned in a sealed tube to prevent the specimen from drying out <pmc-comment>PageBreak</pmc-comment>and X-rays were absorbed by the tissues according to their natural density differences and atomic number. Mineralised structures (jaw apparatus, chaetae) showed up most clearly, as well as the vascular system in the cephalic part of the animal. Muscle tissues are less dense but clearly visible, whereas nervous tissues (brain, ganglia) have almost no X-ray attenuation. Through careful observation, almost all anatomical features can be observed in the data (<xref ref-type="fig" rid="F2">Video 1</xref>); however, external features are partly obscured by a thin ethanol film clinging to the specimen during scanning. By drying the specimen with HMDS, such artefacts can be avoided, since the specimen is fully desiccated while retaining its morphology. (e.g. <xref ref-type="fig" rid="F8">Figs 7</xref>, <xref ref-type="fig" rid="F17">15</xref>). The process removes the liquid medium both from the cells and the surrounding area, resulting in a sharp overall contrast of all tissues. However, density variations between different tissues are less pronounced than in the wet specimen, creating difficulties in distinguishing neighbouring organs (<xref ref-type="fig" rid="F12">Figs 10a</xref>, <xref ref-type="fig" rid="F17">15c–d</xref>). Scanning specimens in air can create other artefacts caused by slight vibration of protruding structures during rotation, such as the chaetae in the parapodium of <italic><named-content content-type="taxon-name">Eunice roussaei</named-content></italic> (<xref ref-type="fig" rid="F8">Fig. 7d</xref>). These artefacts become more pronounced with increasing magnification, since the effect of the movement becomes stronger.</p><p>Iodine and PTA bind to tissues and thus increase their X-ray energy absorption rate. The two stains generally bind to all tissues but exhibited different affinities to certain tissue types, staining them more intensely. Iodine seems to stain calcified struc<pmc-comment>PageBreak</pmc-comment>tures and polysaccharides more strongly, whereas phosphotungstic acid is known to bind to certain proteins (fibrin, collagen) (<xref ref-type="bibr" rid="B84">Quintarelli et al. 1973</xref>) and stains the cuticle and muscle tissues more intensely. In the <italic><named-content content-type="taxon-name">Eunice</named-content></italic> specimen stained with iodine, the cuticle (containing polysaccharides), the jaw apparatus (mineralised with aragonite) and the subacicular hooks were stained strongly compared to other tissues (<xref ref-type="fig" rid="F8">Fig. 7b</xref>). The chemical composition of the subacicular hooks is unknown, but calcified chaetae are known to occur in other polychaetes (e.g. <named-content content-type="taxon-name">Amphinomidae</named-content> (<xref ref-type="bibr" rid="B94">Westheide 1997</xref>), Pogonophora (<xref ref-type="bibr" rid="B35">George and Southward 1973</xref>)), thus their strong staining with iodine could indeed be caused by calcium components. In <italic><named-content content-type="taxon-name">Syllis gracilis</named-content></italic>, the proventricle showed an increased density after staining, here the muscle fibres likewise contain calcium (<xref ref-type="bibr" rid="B13">Briggs et al. 1985</xref>) (<xref ref-type="fig" rid="F16">Fig. 14</xref>). In the <italic><named-content content-type="taxon-name">Eunice</named-content></italic> specimen stained with PTA, the cuticle, which consists of collagen fibres, was stained very intensely, especially in the anterior region of the animal (<xref ref-type="fig" rid="F8">Fig. 7a</xref>). Muscular fibres surrounding the jaw apparatus, the longitudinal muscles and the parapodial muscles are likewise more pronounced with PTA than <pmc-comment>PageBreak</pmc-comment>with iodine (not shown). Contrary to the iodine-stained specimen, the jaw apparatus and the subacicular hooks of the PTA-stained specimen were not stained and remain almost invisible (<xref ref-type="fig" rid="F8">Fig. 7a</xref>). In the two phyllodocid specimens, PTA worked exceptionally well on the pharyngeal apparatus (<xref ref-type="fig" rid="F12">Figs 10</xref><xref ref-type="fig" rid="F13">–11</xref>). In larger specimens, such as in <italic><named-content content-type="taxon-name">Alitta succinea</named-content></italic>, the stain does not easily penetrate into the tissue—black areas in the images indicate unstained tissue (<xref ref-type="fig" rid="F10">Fig. 9f</xref>). PTA penetrates slowly and tissue can bind large volumes of the chemical, so renewal every few days (or larger amounts) of the solution and longer staining times are required for larger specimens.</p></sec><sec sec-type="Visualisation of results"><title>Visualisation of results</title><p>Different approaches to communicate three-dimensional data through a scientific publication have been explored in this study. Firstly, two-dimensional images (screenshots) have been created from the volume rendering software, both normal images and stereo view images which, when viewed with red-cyan glasses, create a 3D-effect. Secondly, videos of interaction with the three-dimensional data have been created and directly embedded into the PDF version of this article. Thirdly, three-dimensional models have been embedded into the PDF which, when viewed with an Adobe Acrobat product, allow the user to interact with them (e.g. rotating, zooming). Both images and videos communicate predefined views of the data. Videos, however, contain a substantially larger amount of information than a single static image. The video of the sequence of cross sections through the data (<xref ref-type="fig" rid="F19">Video 4</xref>) allows the user to investigate the full dataset in a very compact version, thus information is conveyed which would be impossible to include in a publication if only images were used. In other cases, videos allow the viewer to better perceive the spatial relation of structures to each other and understand their relative position and perhaps their functioning in the organism (<xref ref-type="fig" rid="F2">Videos 1</xref><xref ref-type="fig" rid="F11"></xref><xref ref-type="fig" rid="F18">–3</xref>). The interactive models give the reader the greatest freedom to explore the data. Surface models provide an excellent method to present selected information on specific structures (e.g. jaw apparatus, parapodium with chaetae (<xref ref-type="fig" rid="F3">Figs 2</xref><xref ref-type="fig" rid="F9">, 8</xref>)). The resulting models can be rotated, magnified and individual parts can be isolated, allowing the user to explore shapes and spatial relationships from all angles. However, surface models are more suitable for compact structures such as internal organs (see e.g. <xref ref-type="bibr" rid="B87">Ruthensteiner and Hess 2008</xref>) than for the display of fine details. In models with complex shapes and details the number of vertices increased dramatically, leading to very large file sizes which were not suitable for further processing. The relatively complex shape of the parapodium with branchiae is already at the limit of what a current standard desktop computer can process. Smoothing the surface and reduction of vertices reduced the data significantly, but automated smoothing or reduction risks eliminating small but taxonomically important details. The limbate and pectinate chaetae had to be omitted from the model since their thin structures were reduced to random dots along their length as soon as the number of vertices was reduced. The compound chaetae had to be completely remodelled and were finally included as simplified shapes, showing general <pmc-comment>PageBreak</pmc-comment>shape and position but no details of structure. Manual post-processing of the model produced better accounts but requires detailed original data and a good knowledge of the morphology of the specimen. The process can furthermore become very time-consuming and the effort/outcome ratio has to be carefully considered. In the data produced during this study a hybrid approach was used: for most structures manual processing was considered not necessary, some characters were re-modelled as simplified shapes, others omitted. The optimal balance between model size and conveyed information will always depend on the research question being addressed. Software approaches, such as SPIERS (<xref ref-type="bibr" rid="B91">Sutton et al. 2012</xref>) that optimise the rendering of models with a high number of vertices can be an excellent solution if simplified shapes should be presented to the user.</p><fig id="F19" orientation="portrait" position="float"><label>Video 4.</label><caption><p><italic><named-content content-type="taxon-name">Hermodice carunculata</named-content></italic>, sequence of transaxial cross sections from post-pharyngeal chaetigers to the prostomium. Video available for download in full resolution from <ext-link ext-link-type="uri" xlink:href="http://polychaetes.marbigen.org/hermodice-carunculata-slices-micro-ct-video">http://polychaetes.marbigen.org/hermodice-carunculata-slices-micro-ct-video</ext-link> .</p></caption><media xlink:href="ZooKeys-263-001-g019.flv" orientation="portrait" xlink:type="simple" id="d35e1704" position="anchor" mimetype="video" mime-subtype="x-flv"></media></fig><pmc-comment>PageBreak</pmc-comment>
<p>To include the interactive volume data, the dataset had to be substantially reduced in size, resulting in a loss of many details. The embedded data are not a true volume rendering (based on a stack of images) but a pseudo-volume rendering: the software exports images from each angle of the volume-rendered objects and presents these to the user, creating the illusion of a three-dimensional object. The transfer function (and thus the information content) are predefined during model creation and cannot be changed by the end-user. The resulting model has thus not only limitations concerning the available detail of data but also towards the options for the user to explore the data, rotating and zooming being the only options of interactivity. The available transfer functions are furthermore not as sophisticated as in a desktop software. The slight density differences between the muscle tissue of the jaw apparatus and certain structures of the maxillary apparatus (maxillary carriers, accessory lamellae, MV) could not be visualised with the S2PLOT library (<xref ref-type="fig" rid="F6">Fig. 5</xref>). The information value of these embedded objects is therefore limited, and in the present data the surface model is actually able to convey more information than the volume rendering.</p></sec><sec sec-type="Molecular analyses"><title>Molecular analyses</title><p>No differences between the 16s rRNA sequences from samples before and after scanning could be detected; moreover, the 16s rRNA sequences of samples with increasing exposure time to X-ray radiation were also identical. GenBank accession numbers of sequences before scanning and after different radiation energy and exposure time are listed in <xref ref-type="table" rid="T2">Table 2</xref>.</p><table-wrap id="T2" orientation="portrait" position="float"><label>Table 2.</label><caption><p>GenBank accession numbers of the sequences obtained from <italic><named-content content-type="taxon-name">Hediste diversicolor</named-content></italic> specimens before and after scanning.</p></caption><table frame="hsides" rules="groups"><tbody><tr><th rowspan="1" colspan="1"><bold>Sample code</bold></th><th rowspan="1" colspan="1"><bold>Scanning time and voltage</bold></th><th rowspan="1" colspan="1"><bold>GenBank accession number</bold></th></tr><tr><td rowspan="1" colspan="1">NER015</td><td rowspan="1" colspan="1">none (control)</td><td rowspan="1" colspan="1">KC113440</td></tr><tr><td rowspan="1" colspan="1">NER015</td><td rowspan="1" colspan="1">1.5 h, 100 kV</td><td rowspan="1" colspan="1">KC113442</td></tr><tr><td rowspan="1" colspan="1">NER063</td><td rowspan="1" colspan="1">none (control)</td><td rowspan="1" colspan="1">KC113441</td></tr><tr><td rowspan="1" colspan="1">NER063</td><td rowspan="1" colspan="1">12 h, 60 kV</td><td rowspan="1" colspan="1">KC113443</td></tr><tr><td rowspan="1" colspan="1">NER063</td><td rowspan="1" colspan="1">24 h, 60 kV</td><td rowspan="1" colspan="1">KC113444</td></tr><tr><td rowspan="1" colspan="1">NER063</td><td rowspan="1" colspan="1">36 h, 60 kV</td><td rowspan="1" colspan="1">KC113445</td></tr></tbody></table></table-wrap><pmc-comment>PageBreak</pmc-comment>
</sec></sec><sec sec-type="Discussion"><title>Discussion</title><sec sec-type="Is micro-computed tomography suitable for the creation of cybertypes"><title>Is micro-computed tomography suitable for the creation of cybertypes?</title><p><bold>Accuracy and reliability of information:</bold></p><p>Three-dimensional data resulting from micro-CT contain a wealth of information for systematists and taxonomists. The examination of characters in their natural position within the organism allows researchers to assess their true shape but also to infer functionality from morphological structures or even discover new diagnostic characters (e.g. <xref ref-type="bibr" rid="B105">Ziegler et al. 2010</xref>, <xref ref-type="bibr" rid="B106">Zimmermann et al. 2011</xref>). The non-destructive character of the technology allows examining internal features (even of very dense materials) without the need for dissection, leaving the original material intact. At the same time, characters can be investigated truly in three dimensions, whereas dissected material often has to be manipulated or squeezed for its adequate observation under a light microscope, thus rearranging the position of characters. The digital data allow the accurate identification of characters in three dimensions and relate them to each other. If needed, parts of the data can be isolated and examined separately, allowing their free manipulation without other body parts obscuring them (e.g. <xref ref-type="fig" rid="F5">Figs 4b–c</xref>). With appropriate software, measurements can be performed in three dimensions, paving the way for a greater accuracy in these analyses which cannot be obtained with conventional methods.</p><p>Despite these obvious advantages, there are also certain limitations of micro-CT. A crucial point for research on small-sized organisms is image resolution which currently lies—depending on the system—in the range of 0.8 to 100 µm/pixel (although nano-CT systems can reach 0.1 µm/pixel (<xref ref-type="bibr" rid="B93">Wang et al. 2008</xref>)). This is a far coarser resolution than other techniques such as scanning electron microscopy, histological sectioning, cLSM or even light microscopy can achieve. Depending on the size of the specimen and the taxonomically important characters of the taxon, this resolution can prove sufficient. In many taxa, however, fine details in the micrometre range are important characters to distinguish species. These structures might be at the resolution limit of current micro-CT desktop scanners and cannot be adequately displayed. In some cases, the proper observation of characters might also be restricted due to their positioning during the scan. Under a stereo microscope specimens can be twisted or stretched, and obscured features can be made visible by careful manipulation with forceps. The data resulting from X-ray scanning are of course static, since only one position of the animal is imaged in a scan. Virtual dissections or measurements can partly overcome the problems, but in some cases the desired feature might simply not be visible in the data. Another limitation of micro-computed tomography is its inability to detect true colours. Pigmentation patterns can contain valuable taxonomic information in many taxa, but with current systems this information is not available. Recent developments of hybrid systems combining micro-CT with photon counting (<xref ref-type="bibr" rid="B82">Qiong et al. 2012</xref>) show, however, the first promising results of true-colour micro-CT imaging and might become a standard component of desktop scanners in the future. Until such hybrid systems become widely available, a solution for creating true-colour 3D models would be to use photographs or laser scans of the organisms and wrap these around the surface models to create realistic surfaces. However, these options require a substantial amount of manual post-processing and familiarity with 3D creation software.</p><p>Apart from inherent limitations of the technique, the information value of a dataset also depends on a range of parameters and settings during the image production process. Besides artefacts which might be created during image acquisition (<xref ref-type="bibr" rid="B1">Abel et al. 2012</xref>), each step from the acquisition to the final presentation presents the user with a number of settings and choices, each of which can influence the final results (<xref ref-type="fig" rid="F20">Fig. 16</xref>) and cre<pmc-comment>PageBreak</pmc-comment><pmc-comment>PageBreak</pmc-comment>ate problems of comparability between datasets. Since at each step a level of subjectivity is added, the closest approximate to raw data in this chain—the initial volumetric data—are most suitable to represent a virtual type, although the settings during sample preparation, scanning and reconstructing already determine much of the information content. By this definition, derivative data products, such as the interactive surface and volumetric models included in this publication would not qualify for a cybertype. However, the ultimate target which governs these settings and choices is to obtain the optimum result and an adequate view of the character(s) in question. In disciplines other than zoology, different conventions might be more adequate (e.g. <xref ref-type="bibr" rid="B91">Sutton et al. 2012</xref>).</p><fig id="F20" orientation="portrait" position="float"><label>Figure 16.</label><caption><p>Diagram of the image acquisition process from the choice of method to the final presentation of the data, including factors influencing the outcome and information value of the results.</p></caption><graphic xlink:href="ZooKeys-263-001-g020"></graphic></fig><p><bold>Effects on physical specimens:</bold></p><p>Micro-CT is commonly characterised as a non-destructive imaging technique, and indeed neither does the specimen need to be physically manipulated before scanning nor does the exposure to X-ray energies have any visible effect on the morphology of the specimen. However, nothing is known yet about possible tissue damage at the cellular level or after elongated or repeated exposure, so whether the technique is absolutely non-destructive remains to be proved. A certain risk of altering the specimen’s characters lies in their preparation for image acquisition, specifically in methods for contrast enhancement which might irreversibly change tissue characteristics. Although micro-CT does not <italic>per se</italic> require contrast enhancement, such treatment might be necessary, especially when scanning soft-bodied organisms. Tissue staining can lead to excellent results and sharp image contrast (<xref ref-type="bibr" rid="B63">Metscher 2009a</xref>, <xref ref-type="bibr" rid="B64">b</xref>) but no universally applicable protocols for the removal of these stains exist so far. The selectivity of stains towards different tissue types particularly enhances the contrast of certain tissues, whereas signals of other tissues are suppressed and show up less clearly or not at all in the scans. This alteration of natural tissue contrasts renders the specimen potentially useless for future micro-CT examinations with a different purpose. Tissue staining should therefore be used with care, especially if the data are intended for general purpose studies and/or if the material is valuable, since the long-term effects of these chemicals remain yet unknown. On the contrary, specimens scanned in air (either wet or dried) can simply be immersed again into the preservation medium after scanning and are thus available both for imaging methods and for investigation through traditional microscopy techniques, but again not all samples are suitable for this type of treatment. The absolute impact of each method is almost impossible to predict due to the large diversity of tissue types, chemical components and material combinations existing in invertebrates (<xref ref-type="bibr" rid="B33">Faulwetter et al. 2012</xref>). Protocols for best practice have, therefore, still to be identified experimentally for each species. The most suitable method is always determined by a number of factors such as the characteristics of the sample (density, size, shape), the surrounding medium and the scope of the study (<xref ref-type="table" rid="T3">Table 3</xref>). As far as indicated by the results of the current study, X-ray radiation induced by micro-CT seems not to have affected the molecular identity of the specimens, at least no effects of the radiation on the sequenced fragment of the 16S rRNA gene could be detected. Neither exposure to high energies nor repetitive <pmc-comment>PageBreak</pmc-comment>exposure seems to have caused any mutations in this specific part of the molecular material. Previous studies attempting to determine whether exposure of preserved tissue to X-ray radiation causes a fragmentation of the DNA have reached contradicting conclusions (<xref ref-type="bibr" rid="B42">Götherstrom et al. 1995</xref>, <xref ref-type="bibr" rid="B43">Grieshaber et al. 2008</xref>, <xref ref-type="bibr" rid="B73">Paredes et al. 2012</xref>). As <xref ref-type="bibr" rid="B73">Paredes et al. (2012)</xref> point out, the findings of <xref ref-type="bibr" rid="B42">Götherstrom et al. (1995)</xref> and <xref ref-type="bibr" rid="B43">Grieshaber et al. (2008)</xref> might however be biased by the setup of the experiment. <xref ref-type="bibr" rid="B42">Götherstrom et al. (1995)</xref> examined the effect of X-ray radiation on the ability to amplify DNA fragments from pig bones, basing their conclusions on the results on the differences of PCR products brightness in an agarose gel. Results of <xref ref-type="bibr" rid="B43">Grieshaber et al. (2008)</xref> did not show any significant effect of the radiation on the DNA amplification with RT-PCR; nevertheless both studies conclude that exposure to radiation had caused degradation of the DNA. <xref ref-type="bibr" rid="B73">Paredes et al. (2012)</xref>, by comparing pre- and post-CT DNA fragmentation profiles of preserved bird skin, did not detect significant differences of DNA quality before and after scanning, concluding that no quantifiable DNA fragmentation was induced by exposure of the sample to X-ray. However, all <pmc-comment>PageBreak</pmc-comment>studies use different exposure times and energies as well as different methods to detect the effect of radiation on the genetic material, thus all these results are only indicative. Furthermore, none of the previous studies used sequencing data to compare the X-Ray effect on DNA samples. If the methodology is to be applied more broadly, further experiments are clearly needed. Likewise, the possible effect of staining substances on the molecular material should be tested; however, DNA of specimens subjected to different contrast-enhancement methods could be successfully extracted and amplified in previous experiments (<xref ref-type="bibr" rid="B5">Austin and Dillon 1997</xref>, <xref ref-type="bibr" rid="B33">Faulwetter et al. 2012</xref>). The above aspects of the potential impact of the micro-CT technology on preserved biological material must be entirely resolved before this method is suggested for wide use on the specimens deposited in museums. This holds especially true for type-material of nominal species which is unique by definition.</p><table-wrap id="T3" orientation="portrait" position="float"><label>Table 3.</label><caption><p>Overview of different contrast enhancing techniques, their applications and limitations.</p></caption><table frame="hsides" rules="groups"><tbody><tr><th rowspan="1" colspan="1"><bold>Method</bold></th><th rowspan="1" colspan="1"><bold>Use cases</bold></th><th rowspan="1" colspan="1"><bold>Limitations</bold></th><th rowspan="1" colspan="1"><bold>Reversibility</bold></th></tr><tr><td rowspan="1" colspan="1">Removal of preservation medium, scan in humid state</td><td rowspan="1" colspan="1">Natural contrast between different tissue types should be kept
Specimen cannot be stained
Specimen contains dense parts and only these should be visualised</td><td rowspan="1" colspan="1">Remaining drops of liquid between external features might obscure details
Specimens might dry out if scanning time is too long
External body parts such as branchiae, membranes, might collapse into unrecognisable shapes
Soft tissue inside very dense structures cannot be visualised appropriately</td><td rowspan="1" colspan="1">Immerse into preservation medium </td></tr><tr><td rowspan="1" colspan="1">Drying (HMDS, possibly also critical point drying or freeze drying)</td><td rowspan="1" colspan="1">Specimen contains both dense parts and soft tissue and both should be visualised
Specimen cannot be stained nor scanned in a humid state</td><td rowspan="1" colspan="1">Specimens become fragile, external body parts might break off
Long appendages vibrate during rotation and create blur
Natural contrast between tissues is reduced, individual organs might be difficult to separate</td><td rowspan="1" colspan="1">Immerse in >95% ethanol </td></tr><tr><td rowspan="1" colspan="1">Tissue stains (e.g. iodine, PTA, silver staining)</td><td rowspan="1" colspan="1">Specimen contains both dense parts and soft tissue and both should be visualised
Specimen is very small or fragile
Specimen cannot be removed from liquid medium
Specimen has appendages that could vibrate in air or collapse to body when liquid medium is removed
Only certain tissues should be visualised (selective staining)</td><td rowspan="1" colspan="1">Natural contrast between tissues is reduced, individual organs might be difficult to separate
Tissues stain selectively, some tissues might not show up at all in the image
PTA: large specimens need very long staining times and large amounts of stain
Iodine: soft tissue does not stain well when specimen contains large calcareous structures (e.g. mollusc shells, serpulid tubes) </td><td rowspan="1" colspan="1">unknown</td></tr></tbody></table></table-wrap></sec><sec sec-type="Data access and curation of cybertypes"><title>Data access and curation of cybertypes</title><p>Three-dimensional data can be communicated in various ways and through various media. Their true potential lies, however, in the wealth of information that a full volumetric dataset offers to the skilled researcher. Volumetric datasets can easily reach a size of several gigabytes per dataset (<xref ref-type="table" rid="T1">Table 1</xref>), a fact that poses new challenges concerning the management, archival, backup and dissemination of these data. Currently, the community lacks infrastructures, standards and policies that allow the adequate curation of three-dimensional data (<xref ref-type="bibr" rid="B85">Rowe and Frank 2011</xref>). An urgent priority is the creation of an infrastructure for sharing these data and encouraging their reuse. Although a number of morphological databases exist at present (<xref ref-type="bibr" rid="B105">Ziegler et al. 2010</xref>), only few focus on the curation and visualisation of three-dimensional morphology data (e.g. Digimorph – <ext-link ext-link-type="uri" xlink:href="http://www.digimorph.org">http://www.digimorph.org</ext-link> ), Digital Fish Library – <ext-link ext-link-type="uri" xlink:href="http://www.digitalfishlibrary.org">http://www.digitalfishlibrary.org</ext-link> , <xref ref-type="bibr" rid="B10">Berquist et al. 2012</xref>), and none acts as a broad-scale repository of high-resolution volumetric datasets. Apart from these archives, hundreds of three-dimensional datasets of biological specimens have already been produced but remain inaccessible to the research community (<xref ref-type="bibr" rid="B103">Ziegler et al. 2011a</xref>, <xref ref-type="bibr" rid="B11">Boistel et al. 2011</xref>). The development of standards and protocols for archiving and disseminating three-dimensional data as well as the creation of centralised registers to make the information retrievable remains an immediate priority (<xref ref-type="bibr" rid="B105">Ziegler et al. 2010</xref>, <xref ref-type="bibr" rid="B85">Rowe and Frank 2011</xref>) and is crucial for the future success of these developments. In this context, natural history museums and other large natural collections will have to play a central role, not only by digitising their collections and thus massively producing data (as already exemplified by recent efforts (<xref ref-type="bibr" rid="B89">Smith and Blagoderov 2012</xref>)), but also by taking the lead in the development of standards and software for virtual museum collections and curation of cybertypes (for a discussion on the lack of standard file formats for interchange see <xref ref-type="bibr" rid="B91">Sutton et al. 2012</xref>). This leads to another important issue—the current lack of standards to properly document and exchange volumetric data. Without metadata, datasets are neither retrievable nor interpretable. The medical community has developed the <pmc-comment>PageBreak</pmc-comment>DICOM standard (Digital Imaging and Communications in Medicine, <ext-link ext-link-type="uri" xlink:href="http://dicom.nema.org/">http://dicom.nema.org/</ext-link> ) which contains both format definitions and a communication protocol for the description and exchange of volume data. However, this standard is not as universally implemented as one might expect, with different formats and versions being used by different parties (<xref ref-type="bibr" rid="B66">Mildenberger and Jensch 1999</xref>, <xref ref-type="bibr" rid="B78">Pianykh 2012</xref>). Furthermore, to what extent the DICOM specifications are in compliance with the purposes of the taxonomic community remains to be investigated, thus DICOM might or might not prove to be a suitable format for volumetric data definition and exchange. Another limiting factor for the optimal use of volumetric data is that powerful computing capabilities are required for the visualisation, exploration and analysis of large data sets. Although computing power is becoming increasingly more inexpensive, large-scale analyses and comparisons of datasets will likely be limited to high-performance computing centres. Virtual laboratories providing remote access to these facilities could however give a true boost to widespread usage of these data (<xref ref-type="bibr" rid="B105">Ziegler et al. 2010</xref>). System architectures to provide rapid access to three-dimensional data are already being developed by the informatics research community (e.g. <xref ref-type="bibr" rid="B31">Engel et al. 1999</xref>, <xref ref-type="bibr" rid="B54">Kaupp et al. 2002</xref>, <xref ref-type="bibr" rid="B80">Prohaska et al. 2004</xref>, <xref ref-type="bibr" rid="B21">Congote et al. 2009</xref>, <xref ref-type="bibr" rid="B20">2012</xref>). Finally, a third challenge for the biomedical informatics community is the incorporation of three-dimensional information on web sites. A major bottleneck for the publication of volumetric data is still their large size which has to be transferred to the client for visualisation, but also the lack of native integration of this data into HTML standards. Developments such as the VAXML standard (<xref ref-type="bibr" rid="B91">Sutton et al. 2012</xref>) could potentially become the basis for such integrations. The near future will see several promising developments such as the Arivis WebView browser (<ext-link ext-link-type="uri" xlink:href="http://webview3d.arivis.com/">http://webview3d.arivis.com/</ext-link> ) or Voluminous, the web-based version of Drishti (under development, presentation at <ext-link ext-link-type="uri" xlink:href="https://sites.google.com/site/ozvizworkshop/ozviz-2011">https://sites.google.com/site/ozvizworkshop/ozviz-2011</ext-link> ).</p></sec><sec sec-type="The future of virtual taxonomy"><title>The future of virtual taxonomy</title><p>The increasing availability of accurate, three-dimensional virtual representations of biological specimens offers an exciting range of new research opportunities and will significantly accelerate access to first-hand morphological information, thus helping to overcome one of the major bottlenecks in systematic and taxonomic research: the continuous availability of type material to all potential users simultaneously. At present, although virtual specimens in most cases cannot—and should not—replace physical type material, often the desired information can be obtained from a virtual representation. In this study, none of the scanned specimens comply to the hypothetical requirement for a virtual type to provide as much information as the original material. However, different imaging methods (e.g. photography, nano-CT, MRI, OPT) can be employed to produce complementary datasets, and sophisticated future methods could provide a seamless integration of different datasets, incrementally loading additional data when zooming in or focusing on certain characteristics. The availability <pmc-comment>PageBreak</pmc-comment>of information-rich cybertypes would not only protect the actual type material from loss or damage through careless handling, but would also provide simultaneous access by multiple users to the material. This approach would also provide a way to access collections where local restrictions prevent removing specimens from the institution or country.</p><p>The increased creation of three-dimensional taxonomic data will also inevitably influence the way taxonomic data is published. Embedding data as interactive, three-dimensional objects into publications will likely become a standard to usefully convey information. With the recent amendment of the International Code of Zoological Nomenclature (ICZN) that allows taxonomic treatments to be published exclusively electronically (<xref ref-type="bibr" rid="B49">International Commission on Zoological Nomenclature 2012</xref>), multimedia and interactive data can be embedded directly into the publication. This allows these large datasets to be embedded within the presentation of the paper as a single “coherent scientific report” (<xref ref-type="bibr" rid="B104">Ziegler et al. 2011b</xref>). With these regulations, an important milestone has been reached in the process of transforming taxonomy into a cyber-discipline. However, if the concept of virtual type material types is to be officially established, further changes to the ICZN will have to be made that regulate the use of cybertypes to complement physical type material. Specifically, Article 72, which regulates the type concept in nomenclature, should be amended to include the definition of the concept of a “cybertype”, “e-type” or “virtual type” and provide recommendations upon the nomenclatural status, electronic format, access and longevity of such datasets, as well as ensuring that they are made accessible to the public domain under a licence that ensures open access and encourages the creation of derivative works. Furthermore, as <xref ref-type="bibr" rid="B96">Wheeler et al. (2012)</xref> point out, regulations concerning registry of cybertypes in central access points such as ZooBank should be included. Such regulations will allow entirely new ways of data access to emerge, such as virtual realities (<xref ref-type="bibr" rid="B25">Deligiannidis and Jacob 2005</xref>, <xref ref-type="bibr" rid="B59">Laha et al. 2012</xref>), interactive access to virtual specimens on mobile devices (<xref ref-type="bibr" rid="B51">Johnson et al. 2012</xref>) or holographic representations (<xref ref-type="bibr" rid="B50">Javidi and Tajahuerce 2000</xref>, <xref ref-type="bibr" rid="B65">Midgley and Dunin-Borkowski 2009</xref>, <xref ref-type="bibr" rid="B11">Boistel et al. 2011</xref>).</p><p>Despite certain obvious advantages of 3D-imaging technologies, they will need time to evolve into a widely adopted method. At its core, taxonomy is a very traditional discipline, and commonly, changes are adopted at a slow pace. The aforementioned transition of taxonomy into a more data-centric and electronic discipline will need time so the community can learn how to make the best use of this new type of data and the information it contains, as well as to develop the necessary skills to handle these data. This goes hand in hand with technical obstacles that prevent the method from becoming widely used at the present: access to 3D imaging facilities is, although steadily increasing, still limited and often expensive. Special technical skills are needed to produce and process the data, and even with such expertise, the creation of the final dataset (the cybertype) is still a very time-consuming process. However, these arguments hold true for many new technologies, and it will be for the community to decide whether the information value contained in 3D-datasets will allow the technology to survive and to shape the future direction of taxonomy.</p><pmc-comment>PageBreak</pmc-comment>
</sec></sec><sec sec-type="Conclusions"><title>Conclusions</title><p>Up to now, morphology-based systematics and taxonomy have not been able to keep pace with the rapid developments and data creation that characterise other disciplines. New technologies, such as micro-CT and other imaging techniques, will allow massive, computer-accessible data production. This will, in turn, inspire the development of new tools to manage and analyse these data, allowing large-scale morphology-based phylogenies, semi-automated identifications, the formulation of new systematic hypotheses, and advanced research on novel ways of managing, visualising and publishing data. The combined efforts of humans and new technologies will help the discipline to find its way into the digital age and might trigger its renaissance with an impact rivalling the discoveries of the great naturalist era of the 19<sup>th</sup> century.</p></sec></body><back><ack><title>Acknowledgements</title><p>The authors are grateful for support from the following colleagues: Dr Costas Dounas (Hellenic Centre for Marine Research, Greece), Dr Didier Aurelle (Université de la Méditerranée Centre d’Océanologie de Marseille, France) and Dr Chariton Chintiroglou (Aristotelian University of Thessaloniki, Greece) for providing specimens; Dr Jeroen Hostens (SkyScan, Belgium) and Dr Brian Metscher (Department of Theoretical Biology, University of Vienna, Austria) for support and training in the application of various staining methods and scanning techniques; Dr Lyubomir Penev (PENSOFT, Sofia, Bulgaria) and Dr Gordon Paterson (Natural History Museum of London, UK) for inspiring discussions; Dr Farah Ahmed and Daniel Sykes (Natural History Museum of London, UK) for sharing experiences and for an introduction to Drishti, and finally Dr Antonios Magoulas (Hellenic Centre for Marine Research, Greece) for his continuous support through the MARBIGEN project. The reviewers (Dr Daniel Mietchen, Dr Russel Garwood and one anonymous reviewer) are thanked for their suggestions to improve the manuscript. Financial support was provided by the European Union projects MARBIGEN (FP7-REGPOT-2010-1) and ViBRANT (RI-261532).</p></ack><ref-list><title>References</title><ref id="B1"><mixed-citation publication-type="other"><person-group><name><surname>Abel</surname><given-names>RL</given-names></name><name><surname>Laurini</surname><given-names>CR</given-names></name><name><surname>Richter</surname><given-names>M</given-names></name></person-group> (<year>2012</year>)<source> A palaeobiologist’s guide to “virtual” micro-CT preparation. Palaeontologia Electronica 15: 1–16. <ext-link ext-link-type="uri" xlink:href="http://palaeo-electronica.org/content/pdfs/284.pdf">http://palaeo-electronica.org/content/pdfs/284.pdf</ext-link></source></mixed-citation></ref><ref id="B2"><mixed-citation publication-type="other"><person-group><name><surname>Alba-Tercedor</surname><given-names>J</given-names></name><name><surname>Sánchez-Tocino</surname><given-names>L</given-names></name></person-group> (<year>2011</year>)<source> The use of the SkyScan 1172 high-resolution micro-CT to elucidate if the spicules of the “sea slugs” (Mollusca: Nudibranchia, Opisthobranchia) have a structural or a defensive function. In: MicroCT User Meeting Abstract Book, Leuven (Belgium), April 2011. SkyScan, Kontich, Belgium, 113–121. <ext-link ext-link-type="uri" xlink:href="http://www.skyscan.be/company/UM2011/abstract_22.pdf">http://www.skyscan.be/company/UM2011/abstract_22.pdf</ext-link></source></mixed-citation></ref><pmc-comment>PageBreak</pmc-comment>
<ref id="B3"><mixed-citation publication-type="journal"><person-group><name><surname>Audzijonyte</surname><given-names>A</given-names></name><name><surname>Ovcarenko</surname><given-names>I</given-names></name><name><surname>Bastrop</surname><given-names>R</given-names></name><name><surname>Väinölä</surname><given-names>R</given-names></name></person-group> (<year>2008</year>)<article-title> Two cryptic species of the <italic>Hediste diversicolor</italic> group (Polychaeta, Nereididae) in the Baltic Sea, with mitochondrial signatures of different population histories.</article-title><source>Marine Biology </source><volume>155</volume>: <fpage>599</fpage>-<lpage>612</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1007/s00227-008-1055-3">10.1007/s00227-008-1055-3</ext-link></mixed-citation></ref><ref id="B4"><mixed-citation publication-type="other"><person-group><name><surname>Audouin</surname><given-names>JV</given-names></name><name><surname>Milne-Edwards</surname><given-names>H</given-names></name></person-group> (<year>1834</year>)<source> Recherches pour servir a l’histoire naturelle du littoral de la France, ou recuil de mémoires sur l’anatomie, la physiologie, la classification et les moeurs des animaux de nos côtes, 2. Annélides, Crochard, Paris, 290 pp. <ext-link ext-link-type="uri" xlink:href="http://www.biodiversitylibrary.org/ia/recherchespourse21834audo">http://www.biodiversitylibrary.org/ia/recherchespourse21834audo</ext-link></source></mixed-citation></ref><ref id="B5"><mixed-citation publication-type="journal"><person-group><name><surname>Austin</surname><given-names>AD</given-names></name><name><surname>Dillon</surname><given-names>N</given-names></name></person-group> (<year>1997</year>)<article-title> Extraction and PCR of DNA from parasitoid wasps that have been chemically dried.</article-title><source>Australian Journal of Entomology </source><volume>36</volume>: <fpage>241</fpage>-<lpage>244</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1111/j.1440-6055.1997.tb01461.x">10.1111/j.1440-6055.1997.tb01461.x</ext-link></mixed-citation></ref><ref id="B6"><mixed-citation publication-type="other"><person-group><name><surname>Bakken</surname><given-names>T</given-names></name></person-group> (<year>2004</year>)<source> A revision of Nereidinae (Polychaeta, Nereididae). PhD Thesis, Trondheim, Norway: Norwegian University of Science and Technology. <ext-link ext-link-type="uri" xlink:href="http://ntnu.diva-portal.org/smash/record.jsf?pid=diva2:123502">http://ntnu.diva-portal.org/smash/record.jsf?pid=diva2:123502</ext-link></source></mixed-citation></ref><ref id="B7"><mixed-citation publication-type="journal"><person-group><name><surname>Barnes</surname><given-names>D</given-names></name><name><surname>Fluke</surname><given-names>C</given-names></name><name><surname>Bourke</surname><given-names>P</given-names></name><name><surname>Parry</surname><given-names>OT</given-names></name></person-group> (<year>2006</year>)<article-title> An advanced, three-dimensional plotting library for astronomy.</article-title><source>Publications of the Astronomical Society of Australia </source><volume>23</volume>: <fpage>82</fpage>-<lpage>93</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1071/AS06009">10.1071/AS06009</ext-link></mixed-citation></ref><ref id="B8"><mixed-citation publication-type="journal"><person-group><name><surname>Barroso</surname><given-names>R</given-names></name><name><surname>Klautau</surname><given-names>M</given-names></name><name><surname>Solé-Cava</surname><given-names>AM</given-names></name><name><surname>de Paiva</surname><given-names>PC</given-names></name></person-group> (<year>2009</year>)<article-title><italic>Eurythoe complanata</italic> (Polychaeta: Amphinomidae), the “cosmopolitan” fireworm, consists of at least three cryptic species.</article-title><source>Marine Biology </source><volume>157</volume>: <fpage>69</fpage>-<lpage>80</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1007/s00227-009-1296-9">10.1007/s00227-009-1296-9</ext-link></mixed-citation></ref><ref id="B9"><mixed-citation publication-type="other"><person-group><name><surname>Benson</surname><given-names>DA</given-names></name><name><surname>Karsch-Mizrachi</surname><given-names>I</given-names></name><name><surname>Lipman</surname><given-names>DJ</given-names></name><name><surname>Ostell</surname><given-names>J</given-names></name><name><surname>Wheeler</surname><given-names>DL</given-names></name></person-group> (<year>2005</year>)<source> GenBank. Nucleic Acids Research 33: D34-D38. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1093/nar/gki063">10.1093/nar/gki063</ext-link></mixed-citation></ref><ref id="B10"><mixed-citation publication-type="other"><person-group><name><surname>Berquist</surname><given-names>RM</given-names></name><name><surname>Gledhill</surname><given-names>KM</given-names></name><name><surname>Peterson</surname><given-names>MW</given-names></name><name><surname>Doan</surname><given-names>AH</given-names></name><name><surname>Baxter</surname><given-names>GT</given-names></name><name><surname>Yopak</surname><given-names>KE</given-names></name><name><surname>Kang</surname><given-names>N</given-names></name><name><surname>Walker</surname><given-names>HJ</given-names></name><name><surname>Hastings</surname><given-names>PA</given-names></name><name><surname>Frank</surname><given-names>LR</given-names></name></person-group> (<year>2012</year>)<source> The Digital Fish Library: Using MRI to Digitize, Database, and Document the Morphological Diversity of Fish. PLoS ONE 7: e34499. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1371/journal.pone.0034499">10.1371/journal.pone.0034499</ext-link></mixed-citation></ref><ref id="B11"><mixed-citation publication-type="journal"><person-group><name><surname>Boistel</surname><given-names>R</given-names></name><name><surname>Swoger</surname><given-names>J</given-names></name><name><surname>Kržič</surname><given-names>U</given-names></name><name><surname>Fernandez</surname><given-names>V</given-names></name><name><surname>Gillet</surname><given-names>B</given-names></name><name><surname>Reynaud</surname><given-names>EG</given-names></name></person-group> (<year>2011</year>)<article-title> The future of three-dimensional microscopic imaging in marine biology.</article-title><source>Marine Ecology </source><volume>32</volume>: <fpage>438</fpage>-<lpage>452</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1111/j.1439-0485.2011.00442.x">10.1111/j.1439-0485.2011.00442.x</ext-link></mixed-citation></ref><ref id="B12"><mixed-citation publication-type="other"><person-group><name><surname>Bond</surname><given-names>J</given-names></name><name><surname>Hörnschmeyer</surname><given-names>T</given-names></name><name><surname>Young</surname><given-names>P</given-names></name><name><surname>Yang</surname><given-names>L</given-names></name></person-group> (<year>2008</year>)<source> Meshing of XMT data to study the functional morphology of beetle <italic>Priacma serrata</italic>’s head and mandible. In: Proceedings of the 2nd International Conference of Bionic Engineering (ICBE’08), Changchun (China) October 2008. Jilin University, Changchun, 1–11.</source></mixed-citation></ref><ref id="B13"><mixed-citation publication-type="journal"><person-group><name><surname>Briggs</surname><given-names>RT</given-names></name><name><surname>Chaffee</surname><given-names>JE</given-names></name><name><surname>Anderson</surname><given-names>M</given-names></name></person-group> (<year>1985</year>)<article-title> Calcium-containgn granules in myoepithelial cells of the polychaete <italic>Syllis spongiphila</italic>: Possible ionic modulators.</article-title><source>Tissue </source><volume>17</volume>: <fpage>923</fpage>-<lpage>928</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/0040-8166(85)90046-1">10.1016/0040-8166(85)90046-1</ext-link></mixed-citation></ref><ref id="B14"><mixed-citation publication-type="other"><person-group><name><surname>Budd</surname><given-names>GE</given-names></name><name><surname>Olsson</surname><given-names>L</given-names></name></person-group> (<year>2007</year>)<source> A renaissance for evolutionary morphology. Acta Zoologica 88: 1. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1111/j.1463-6395.2007.00262.x">10.1111/j.1463-6395.2007.00262.x</ext-link></mixed-citation></ref><ref id="B15"><mixed-citation publication-type="journal"><person-group><name><surname>Böggemann</surname><given-names>M</given-names></name></person-group> (<year>2002</year>)<article-title> Revision of the Glyceridae Grube 1850 (Annelida: Polychaeta).</article-title><source>Abhandlungen der Senckenbergischen Naturforschenden Gesellschaft </source><volume>555</volume>: <fpage>1</fpage>-<lpage>249</lpage></mixed-citation></ref><ref id="B16"><mixed-citation publication-type="other"><person-group><name><surname>Carrera-Parra</surname><given-names>LF</given-names></name></person-group> (<year>2006a</year>)<source> Revision of <italic>Lumbrineris</italic> de Blainville, 1828 (Polychaeta: Lumbrineridae). Zootaxa 1336: 1–64. <ext-link ext-link-type="uri" xlink:href="https://www.scopus.com/record/display.url?eid=2-s2.0-33750323441&origin=resultslist">https://www.scopus.com/record/display.url?eid=2-s2.0-33750323441&origin=resultslist</ext-link></source></mixed-citation></ref><pmc-comment>PageBreak</pmc-comment>
<ref id="B17"><mixed-citation publication-type="other"><person-group><name><surname>Carrera-Parra</surname><given-names>LF</given-names></name></person-group> (<year>2006b</year>)<source> Phylogenetic analysis of Lumbrineridae Schmarda, 1861 (Annelida: Polychaeta). Zootaxa 36: 1–36. <ext-link ext-link-type="uri" xlink:href="https://www.scopus.com/record/display.url?eid=2-s2.0-33750375288&origin=resultslist">https://www.scopus.com/record/display.url?eid=2-s2.0-33750375288&origin=resultslist</ext-link></source></mixed-citation></ref><ref id="B18"><mixed-citation publication-type="journal"><person-group><name><surname>Carvalho</surname><given-names>MR</given-names></name><name><surname>Bockmann</surname><given-names>FA</given-names></name><name><surname>Amorim</surname><given-names>DS</given-names></name><name><surname>Brandão</surname><given-names>CRF</given-names></name><name><surname>Vivo</surname><given-names>M</given-names></name><name><surname>Figueiredo</surname><given-names>JL</given-names></name><name><surname>Britski</surname><given-names>HA</given-names></name><name><surname>Pinna</surname><given-names>MCC</given-names></name><name><surname>Menezes</surname><given-names>NA</given-names></name><name><surname>Marques</surname><given-names>FPL</given-names></name><name><surname>Papavero</surname><given-names>N</given-names></name><name><surname>Cancello</surname><given-names>EM</given-names></name><name><surname>Crisci</surname><given-names>JV</given-names></name><name><surname>McEachran</surname><given-names>JD</given-names></name><name><surname>Schelly</surname><given-names>RC</given-names></name><name><surname>Lundberg</surname><given-names>JG</given-names></name><name><surname>Gill</surname><given-names>AC</given-names></name><name><surname>Britz</surname><given-names>R</given-names></name><name><surname>Wheeler</surname><given-names>QD</given-names></name><name><surname>Stiassny</surname><given-names>MLJ</given-names></name><name><surname>Parenti</surname><given-names>LR</given-names></name><name><surname>Page</surname><given-names>LM</given-names></name><name><surname>Wheeler</surname><given-names>WC</given-names></name><name><surname>Faivovich</surname><given-names>J</given-names></name><name><surname>Vari</surname><given-names>RP</given-names></name><name><surname>Grande</surname><given-names>L</given-names></name><name><surname>Humphries</surname><given-names>CJ</given-names></name><name><surname>DeSalle</surname><given-names>R</given-names></name><name><surname>Ebach</surname><given-names>MC</given-names></name><name><surname>Nelson</surname><given-names>GJ</given-names></name></person-group> (<year>2007</year>)<article-title> Taxonomic Impediment or Impediment to Taxonomy? A Commentary on Systematics and the Cybertaxonomic-Automation Paradigm.</article-title><source>Evolutionary Biology </source><volume>34</volume>: <fpage>140</fpage>-<lpage>143</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1007/s11692-007-9011-6">10.1007/s11692-007-9011-6</ext-link></mixed-citation></ref><ref id="B19"><mixed-citation publication-type="journal"><person-group><name><surname>Claparède</surname><given-names>É</given-names></name></person-group> (<role>1870</role>) <source>Les annélides chétopodes du Golfe de Naples. Supplément. Mémoires de la Société de physique et d’histoire naturelle de Genève</source><volume>20</volume>: <fpage>365</fpage>–<lpage>542</lpage><ext-link ext-link-type="uri" xlink:href="http://www.biodiversitylibrary.org/item/18576">http://www.biodiversitylibrary.org/item/18576</ext-link></mixed-citation></ref><ref id="B20"><mixed-citation publication-type="other"><person-group><name><surname>Congote</surname><given-names>J</given-names></name><name><surname>Kabongo</surname><given-names>L</given-names></name><name><surname>Moreno</surname><given-names>A</given-names></name><name><surname>Segura</surname><given-names>A</given-names></name><name><surname>Beristain</surname><given-names>A</given-names></name><name><surname>Posada</surname><given-names>J</given-names></name><name><surname>Ruiz</surname><given-names>O</given-names></name></person-group> (<year>2012</year>)<article-title> Volume Ray Casting in WebGL.</article-title> In: <person-group><name><surname>Mukai</surname><given-names>N</given-names></name></person-group> (<role>Ed</role>). <issue-title>Computer Graphics.</issue-title><source>InTech, Rijeka</source>: <fpage>157</fpage>-<lpage>178</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.5772/34878">10.5772/34878</ext-link></mixed-citation></ref><ref id="B21"><mixed-citation publication-type="other"><person-group><name><surname>Congote</surname><given-names>J</given-names></name><name><surname>Kabongo</surname><given-names>L</given-names></name><name><surname>Moreno</surname><given-names>A</given-names></name><name><surname>Segura</surname><given-names>A</given-names></name><name><surname>Posada</surname><given-names>J</given-names></name><name><surname>Ruiz</surname><given-names>O</given-names></name></person-group> (<year>2009</year>)<source> Interactive visualization of volumetric data with WebGL in real-time. In: Web3D ’11 Proceedings of the 16th International Conference on 3D Web Technology, Paris (France), June 2011. ACM New York, 137–146. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1145/2010425.2010449">10.1145/2010425.2010449</ext-link></mixed-citation></ref><ref id="B22"><mixed-citation publication-type="other"><person-group><name><surname>Cook</surname><given-names>LG</given-names></name><name><surname>Edwards</surname><given-names>RD</given-names></name><name><surname>Crisp</surname><given-names>MD</given-names></name><name><surname>Hardy</surname><given-names>NB</given-names></name></person-group> (<year>2010</year>)<source> Need morphology always be required for new species descriptions? Invertebrate Systematics 24: 322–326. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1071/IS10011">10.1071/IS10011</ext-link></mixed-citation></ref><ref id="B23"><mixed-citation publication-type="other"><person-group><name><surname>Csösz</surname><given-names>S</given-names></name></person-group> (<year>2012</year>)<source> Nematode infection as significant source of unjustified taxonomic descriptions in ants (Hymenoptera: Formicidae). Myrmecological News 17: 27–31. <ext-link ext-link-type="uri" xlink:href="http://www.myrmecologicalnews.org/cms/images/pdf/volume17/mn17_27-31_non-printable.pdf">http://www.myrmecologicalnews.org/cms/images/pdf/volume17/mn17_27-31_non-printable.pdf</ext-link></source></mixed-citation></ref><ref id="B24"><mixed-citation publication-type="journal"><person-group><name><surname>Deans</surname><given-names>AR</given-names></name><name><surname>Yoder</surname><given-names>MJ</given-names></name><name><surname>Balhoff</surname><given-names>JP</given-names></name></person-group> (<year>2011</year>)<article-title> Time to change how we describe biodiversity.</article-title><source>Trends in Ecology & Evolution </source><volume>27</volume>: <fpage>78</fpage>-<lpage>84</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.tree.2011.11.007">10.1016/j.tree.2011.11.007</ext-link><pub-id pub-id-type="pmid">22189359</pub-id></mixed-citation></ref><ref id="B25"><mixed-citation publication-type="other"><person-group><name><surname>Deligiannidis</surname><given-names>L</given-names></name><name><surname>Jacob</surname><given-names>RJK</given-names></name></person-group> (<year>2005</year>)<source> An Immersive Environment for the Vase Museum. In: HCI’05 International Conference on Human-Computer Interaction, Las Vegas (USA), July 2005. HCI International, 54–62. <ext-link ext-link-type="uri" xlink:href="https://www.scopus.com/record/display.url?eid=2-s2.0-60749101560&origin=resultslist">https://www.scopus.com/record/display.url?eid=2-s2.0-60749101560&origin=resultslist</ext-link></source></mixed-citation></ref><ref id="B26"><mixed-citation publication-type="journal"><person-group><name><surname>Dierick</surname><given-names>M</given-names></name><name><surname>Cnudde</surname><given-names>V</given-names></name><name><surname>Masschaele</surname><given-names>B</given-names></name><name><surname>Vlassenbroeck</surname><given-names>J</given-names></name><name><surname>Van Hoorebeke</surname><given-names>L</given-names></name><name><surname>Jacobs</surname><given-names>P</given-names></name></person-group> (<year>2007</year>)<article-title> Micro-CT of fossils preserved in amber.</article-title><source>Nuclear Instruments and Methods in Physics Research A </source><volume>580</volume>: <fpage>641</fpage>-<lpage>643</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.nima.2007.05.030">10.1016/j.nima.2007.05.030</ext-link></mixed-citation></ref><ref id="B27"><mixed-citation publication-type="journal"><person-group><name><surname>Dinley</surname><given-names>J</given-names></name><name><surname>Hawkins</surname><given-names>L</given-names></name><name><surname>Paterson</surname><given-names>GLJ</given-names></name><name><surname>Ball</surname><given-names>AD</given-names></name><name><surname>Sinclair</surname><given-names>I</given-names></name><name><surname>Sinnett-Jones</surname><given-names>P</given-names></name><name><surname>Lanham</surname><given-names>S</given-names></name></person-group> (<year>2010</year>)<article-title> Micro-computed X-ray tomography: a new non-destructive method of assessing sectional, fly-through and 3D imaging of a soft-bodied marine worm.</article-title><source>Journal of Microscopy </source><volume>238</volume>: <fpage>123</fpage>-<lpage>33</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1111/j.1365-2818.2009.03335.x">10.1111/j.1365-2818.2009.03335.x</ext-link><pub-id pub-id-type="pmid">20529060</pub-id></mixed-citation></ref><ref id="B28"><mixed-citation publication-type="journal"><person-group><name><surname>Dunlop</surname><given-names>JA</given-names></name><name><surname>Wirth</surname><given-names>S</given-names></name><name><surname>Penney</surname><given-names>D</given-names></name><name><surname>McNeil</surname><given-names>A</given-names></name><name><surname>Bradley</surname><given-names>RS</given-names></name><name><surname>Withers</surname><given-names>PJ</given-names></name><name><surname>Preziosi</surname><given-names>RF</given-names></name></person-group> (<year>2011</year>)<article-title> A minute fossil phoretic mite recovered by phase-contrast X-ray computed tomography.</article-title><source>Biology letters </source><volume>8</volume>: <fpage>457</fpage>-<lpage>460</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1098/rsbl.2011.0923">10.1098/rsbl.2011.0923</ext-link><pub-id pub-id-type="pmid">22072283</pub-id></mixed-citation></ref><ref id="B29"><mixed-citation publication-type="journal"><person-group><name><surname>Dunn</surname><given-names>CW</given-names></name><name><surname>Hejnol</surname><given-names>A</given-names></name><name><surname>Matus</surname><given-names>DQ</given-names></name><name><surname>Pang</surname><given-names>K</given-names></name><name><surname>Browne</surname><given-names>WE</given-names></name><name><surname>Smith</surname><given-names>SA</given-names></name><name><surname>Seaver</surname><given-names>E</given-names></name><name><surname>Rouse</surname><given-names>GW</given-names></name><name><surname>Obst</surname><given-names>M</given-names></name><name><surname>Edgecombe</surname><given-names>GD</given-names></name><name><surname>Sørensen</surname><given-names>MV</given-names></name><name><surname>Haddock</surname><given-names>SHD</given-names></name><name><surname>Schmidt-Rhaesa</surname><given-names>A</given-names></name><name><surname>Okusu</surname><given-names>A</given-names></name><name><surname>Kristensen</surname><given-names>RM</given-names></name><name><surname>Wheeler</surname><given-names>WC</given-names></name><name><surname>Martindale</surname><given-names>MQ</given-names></name><name><surname>Giribet</surname><given-names>G</given-names></name></person-group> (<year>2008</year>)<article-title> Broad phylogenomic sampling improves resolution of the animal tree of life.</article-title><source>Nature </source><volume>452</volume>: <fpage>745</fpage>-<lpage>749</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1038/nature06614">10.1038/nature06614</ext-link><pub-id pub-id-type="pmid">18322464</pub-id></mixed-citation></ref><ref id="B30"><mixed-citation publication-type="journal"><person-group><name><surname>Edgecombe</surname><given-names>GD</given-names></name><name><surname>Giribet</surname><given-names>G</given-names></name><name><surname>Dunn</surname><given-names>CW</given-names></name><name><surname>Hejnol</surname><given-names>A</given-names></name><name><surname>Kristensen</surname><given-names>RM</given-names></name><name><surname>Neves</surname><given-names>RC</given-names></name><name><surname>Rouse</surname><given-names>GW</given-names></name><name><surname>Worsaae</surname><given-names>K</given-names></name><name><surname>Sørensen</surname><given-names>MV</given-names></name></person-group> (<year>2011</year>)<article-title> Higher-level metazoan relationships: recent progress and remaining questions.</article-title><source>Organisms Diversity & Evolution </source><volume>11</volume>: <fpage>151</fpage>-<lpage>172</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1007/s13127-011-0044-4">10.1007/s13127-011-0044-4</ext-link></mixed-citation></ref><ref id="B31"><mixed-citation publication-type="other"><person-group><name><surname>Engel</surname><given-names>K</given-names></name><name><surname>Sommer</surname><given-names>O</given-names></name><name><surname>Ernst</surname><given-names>C</given-names></name><name><surname>Ertl</surname><given-names>T</given-names></name></person-group> (<year>1999</year>)<source> Remote 3D Visualization using Image-Streaming Techniques. In: Proceedings of the 11th International Conference on Systems Research, Informatics and Cybernetics, Baden-Baden (Germany), August 1999. International Institute for Advanced Studies in Systems Research and Cybernetics, 91–96.</source></mixed-citation></ref><ref id="B32"><mixed-citation publication-type="other"><person-group><name><surname>Evenhuis</surname><given-names>NL</given-names></name></person-group> (<year>2007</year>)<source> Helping Solve the “other” Taxonomic Impediment: Completing the Eight Steps to Total Enlightenment and Taxonomic Nirvana. Zootaxa 1407: 3–12. <ext-link ext-link-type="uri" xlink:href="https://www.scopus.com/record/display.url?eid=2-s2.0-33847357817&origin=resultslist">https://www.scopus.com/record/display.url?eid=2-s2.0-33847357817&origin=resultslist</ext-link></source></mixed-citation></ref><ref id="B33"><mixed-citation publication-type="other"><person-group><name><surname>Faulwetter</surname><given-names>S</given-names></name><name><surname>Dailianis</surname><given-names>T</given-names></name><name><surname>Vasileiadou</surname><given-names>A</given-names></name><name><surname>Arvanitidis</surname><given-names>C</given-names></name></person-group> (<year>2012</year>)<source> Investigation of contrast enhancing techniques for the application of Micro-CT in marine biodiversity studies. In: SkyScan MicroCT User meeting 2012. SkyScan, Brussels, Belgium, 12–20. <ext-link ext-link-type="uri" xlink:href="http://www.skyscan.be/company/UM2012/02.pdf">http://www.skyscan.be/company/UM2012/02.pdf</ext-link></source></mixed-citation></ref><ref id="B34"><mixed-citation publication-type="journal"><person-group><name><surname>Folmer</surname><given-names>O</given-names></name><name><surname>Black</surname><given-names>M</given-names></name><name><surname>Hoeh</surname><given-names>W</given-names></name><name><surname>Lutz</surname><given-names>R</given-names></name><name><surname>Vrijenhoek</surname><given-names>R</given-names></name></person-group> (<year>1994</year>)<article-title> DNA primers for amplification of mitochondrial cytochrome c oxidase unit I from diverse metazoan invertebrates.</article-title><source>Molecular Marine Biology and Biotechnology </source><volume>3</volume>: <fpage>294</fpage>-<lpage>299</lpage><pub-id pub-id-type="pmid">7881515</pub-id></mixed-citation></ref><ref id="B35"><mixed-citation publication-type="journal"><person-group><name><surname>George</surname><given-names>JD</given-names></name><name><surname>Southward</surname><given-names>EC</given-names></name></person-group> (<year>1973</year>)<article-title> A comparative study of the setae of Pogonophora and polychaetous Annelida.</article-title><source>Journal of the Marine Biological Association of the United Kingdom </source><volume>53</volume>: <fpage>397</fpage>-<lpage>401</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1017/S0025315400022359">10.1017/S0025315400022359</ext-link></mixed-citation></ref><ref id="B36"><mixed-citation publication-type="journal"><person-group><name><surname>Giribet</surname><given-names>G</given-names></name></person-group> (<year>2010</year>)<article-title> A new dimension in combining data? The use of morphology and phylogenomic data in metazoan systematics.</article-title><source>Acta Zoologica </source><volume>91</volume>: <fpage>11</fpage>-<lpage>19</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1111/j.1463-6395.2009.00420.x">10.1111/j.1463-6395.2009.00420.x</ext-link></mixed-citation></ref><ref id="B37"><mixed-citation publication-type="journal"><person-group><name><surname>Godfray</surname><given-names>HCJ</given-names></name></person-group> (<year>2002a</year>)<article-title> Challenges for taxonomy.</article-title><source>Nature </source><volume>417</volume>: <fpage>17</fpage>-<lpage>19</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1038/417017a">10.1038/417017a</ext-link><pub-id pub-id-type="pmid">11986643</pub-id></mixed-citation></ref><ref id="B38"><mixed-citation publication-type="other"><person-group><name><surname>Godfray</surname><given-names>HCJ</given-names></name></person-group> (<year>2002b</year>)<source> Towards taxonomy’s “glorious revolution”. Nature 420: 461. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1038/420461a">10.1038/420461a</ext-link></mixed-citation></ref><ref id="B39"><mixed-citation publication-type="journal"><person-group><name><surname>Godfray</surname><given-names>HCJ</given-names></name></person-group> (<year>2007</year>)<article-title> Linnaeus in the information age.</article-title><source>Nature </source><volume>446</volume>: <fpage>259</fpage>-<lpage>260</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1038/446259a">10.1038/446259a</ext-link><pub-id pub-id-type="pmid">17361160</pub-id></mixed-citation></ref><ref id="B40"><mixed-citation publication-type="other"><person-group><name><surname>Golding</surname><given-names>RE</given-names></name><name><surname>Jones</surname><given-names>AS</given-names></name></person-group> (<year>2006</year>)<source> Micro-CT as a novel technique for 3D reconstruction of molluscan anatomy. Molluscan Research 27: 123–128. <ext-link ext-link-type="uri" xlink:href="https://www.scopus.com/record/display.url?eid=2-s2.0-36048997602&origin=resultslist">https://www.scopus.com/record/display.url?eid=2-s2.0-36048997602&origin=resultslist</ext-link></source></mixed-citation></ref><ref id="B41"><mixed-citation publication-type="journal"><person-group><name><surname>Golding</surname><given-names>RE</given-names></name><name><surname>Ponder</surname><given-names>WF</given-names></name><name><surname>Byrne</surname><given-names>M</given-names></name></person-group> (<year>2009</year>)<article-title> Three-dimensional reconstruction of the odontophoral cartilages of Caenogastropoda (Mollusca: Gastropoda) using micro-CT: Morphology and phylogenetic significance.</article-title><source>Journal of Morphology </source><volume>270</volume>: <fpage>558</fpage>-<lpage>587</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1002/jmor.10699">10.1002/jmor.10699</ext-link><pub-id pub-id-type="pmid">19107810</pub-id></mixed-citation></ref><ref id="B42"><mixed-citation publication-type="journal"><person-group><name><surname>Götherstrom</surname><given-names>A</given-names></name><name><surname>Fischer</surname><given-names>C</given-names></name><name><surname>Linden</surname><given-names>K</given-names></name></person-group> (<year>1995</year>)<article-title> X-raying ancient bone: a destructive method in connection with DNA analysis.</article-title><source>Laborativ Arkeologi </source><volume>8</volume>: <fpage>26</fpage>-<lpage>28</lpage></mixed-citation></ref><ref id="B43"><mixed-citation publication-type="journal"><person-group><name><surname>Grieshaber</surname><given-names>BM</given-names></name><name><surname>Osborne</surname><given-names>DL</given-names></name><name><surname>Doubleday</surname><given-names>AF</given-names></name><name><surname>Kaestle</surname><given-names>FA</given-names></name></person-group> (<year>2008</year>)<article-title> A pilot study into the effects of X-ray and computed tomography exposure on the amplification of DNA from bone.</article-title><source>Journal of Archaeological Science </source><volume>35</volume>: <fpage>681</fpage>-<lpage>687</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.jas.2007.06.001">10.1016/j.jas.2007.06.001</ext-link></mixed-citation></ref><pmc-comment>PageBreak</pmc-comment>
<ref id="B44"><mixed-citation publication-type="other"><person-group><name><surname>Grube</surname><given-names>AE</given-names></name></person-group> (<year>1840</year>)<source> Actinien, Echinodermen und Würmer des Adriatischen- und Mittelmeers, nach eigenen Sammlungen beschrieben. J.H. Bon, Königsberg, 92 pp. <ext-link ext-link-type="uri" xlink:href="http://www.biodiversitylibrary.org/bibliography/10133">http://www.biodiversitylibrary.org/bibliography/10133</ext-link></source></mixed-citation></ref><ref id="B45"><mixed-citation publication-type="other"><person-group><name><surname>Heim</surname><given-names>I</given-names></name><name><surname>Nickel</surname><given-names>M</given-names></name></person-group> (<year>2010</year>)<source> Description and molecular phylogeny of <italic>Tethya leysae</italic> sp. nov. (Porifera, Demospongiae, Hadromerida) from the Canadian Northeast Pacific with remarks on the use of microtomography in sponge taxonomy. Zootaxa 21: 1–21. <ext-link ext-link-type="uri" xlink:href="https://www.scopus.com/record/display.url?eid=2-s2.0-77953923313&origin=resultslist">https://www.scopus.com/record/display.url?eid=2-s2.0-77953923313&origin=resultslist</ext-link></source></mixed-citation></ref><ref id="B46"><mixed-citation publication-type="other"><person-group><name><surname>Hendrickx</surname><given-names>H</given-names></name><name><surname>Cnudde</surname><given-names>V</given-names></name><name><surname>Masschaele</surname><given-names>B</given-names></name><name><surname>Dierick</surname><given-names>M</given-names></name><name><surname>Vlassenbroeck</surname><given-names>J</given-names></name><name><surname>Van Hoorebeke</surname><given-names>L</given-names></name></person-group> (<year>2006</year>)<source> Description of a new fossil Pseudogarypus (Pseudoscorpiones: Pseudogarypidae) with the use of X-ray micro-CT to penetrate opaque amber. Zootaxa 1305: 41–50. <ext-link ext-link-type="uri" xlink:href="https://www.scopus.com/record/display.url?eid=2-s2.0-33748599256&origin=resultslist">https://www.scopus.com/record/display.url?eid=2-s2.0-33748599256&origin=resultslist</ext-link></source></mixed-citation></ref><ref id="B47"><mixed-citation publication-type="journal"><person-group><name><surname>Holford</surname><given-names>P</given-names></name></person-group> (<year>2008</year>)<article-title> X-ray computerised microtomography (MicroCT): a new technique for assessing external and internal morphology of bees.</article-title><source>Journal of Apicultural Research </source><volume>47</volume>: <fpage>286</fpage>-<lpage>291</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.3896/IBRA.1.47.4.09">10.3896/IBRA.1.47.4.09</ext-link></mixed-citation></ref><ref id="B48"><mixed-citation publication-type="journal"><person-group><name><surname>Huckstorf</surname><given-names>K</given-names></name><name><surname>Wirkner</surname><given-names>CS</given-names></name></person-group> (<year>2011</year>)<article-title> Comparative morphology of the hemolymph vascular system in krill (Euphausiacea; Crustacea).</article-title><source>Arthropod structure & development </source><volume>40</volume>: <fpage>39</fpage>-<lpage>53</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.asd.2010.07.004">10.1016/j.asd.2010.07.004</ext-link><pub-id pub-id-type="pmid">20678584</pub-id></mixed-citation></ref><ref id="B49"><mixed-citation publication-type="journal"><person-group><name><surname>International</surname><given-names>Commission on Zoological Nomenclature</given-names></name></person-group> (<year>2012</year>)<article-title> Amendment of Articles 8, 9, 10, 21 and 78 of the International Code of Zoological Nomenclature to expand and refine methods of publication.</article-title><source>ZooKeys </source><volume>219</volume>: <fpage>1</fpage>-<lpage>10</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.3897/zookeys.219.3944">10.3897/zookeys.219.3944</ext-link><pub-id pub-id-type="pmid">22977348</pub-id></mixed-citation></ref><ref id="B50"><mixed-citation publication-type="journal"><person-group><name><surname>Javidi</surname><given-names>B</given-names></name><name><surname>Tajahuerce</surname><given-names>E</given-names></name></person-group> (<year>2000</year>)<article-title> Three-dimensional object recognition by use of digital holography.</article-title><source>Optics Letters </source><volume>25</volume>: <fpage>610</fpage>-<lpage>612</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1364/OL.25.000610">10.1364/OL.25.000610</ext-link><pub-id pub-id-type="pmid">18064126</pub-id></mixed-citation></ref><ref id="B51"><mixed-citation publication-type="journal"><person-group><name><surname>Johnson</surname><given-names>PT</given-names></name><name><surname>Zimmerman</surname><given-names>SL</given-names></name><name><surname>Heath</surname><given-names>D</given-names></name><name><surname>Eng</surname><given-names>J</given-names></name><name><surname>Horton</surname><given-names>KM</given-names></name><name><surname>Scott</surname><given-names>WW</given-names></name><name><surname>Fishman</surname><given-names>EK</given-names></name></person-group> (<year>2012</year>)<article-title> The iPad as a mobile device for CT display and interpretation: diagnostic accuracy for identification of pulmonary embolism.</article-title><source>Emergency Radiology </source><volume>19</volume>: <fpage>323</fpage>-<lpage>327</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1007/s10140-012-1037-0">10.1007/s10140-012-1037-0</ext-link><pub-id pub-id-type="pmid">22450843</pub-id></mixed-citation></ref><ref id="B52"><mixed-citation publication-type="journal"><person-group><name><surname>Joppa</surname><given-names>LN</given-names></name><name><surname>Roberts</surname><given-names>DL</given-names></name><name><surname>Pimm</surname><given-names>SL</given-names></name></person-group> (<year>2011a</year>)<article-title> The population ecology and social behaviour of taxonomists.</article-title><source>Trends in Ecology & Evolution </source><volume>26</volume>: <fpage>551</fpage>-<lpage>553</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.tree.2011.07.010">10.1016/j.tree.2011.07.010</ext-link><pub-id pub-id-type="pmid">21862170</pub-id></mixed-citation></ref><ref id="B53"><mixed-citation publication-type="other"><person-group><name><surname>Joppa</surname><given-names>LN</given-names></name><name><surname>Roberts</surname><given-names>DL</given-names></name><name><surname>Pimm</surname><given-names>SL</given-names></name></person-group> (<year>2011b</year>)<source> Taxonomy that matters: Response to Bacher. Trends in Ecology & Evolution 27: 66. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.tree.2011.11.003">10.1016/j.tree.2011.11.003</ext-link></mixed-citation></ref><ref id="B54"><mixed-citation publication-type="other"><person-group><name><surname>Kaupp</surname><given-names>G</given-names></name><name><surname>Stepanenko</surname><given-names>S</given-names></name><name><surname>Herrmann</surname><given-names>A</given-names></name></person-group> (<year>2002</year>)<source> World-wide interaction with 3D-data. In: SSGRR2002S, L’Aquila (Italy), July/August 2002. SSGRR, 1–11.</source></mixed-citation></ref><ref id="B55"><mixed-citation publication-type="journal"><person-group><name><surname>Kersemans</surname><given-names>V</given-names></name><name><surname>Thompson</surname><given-names>J</given-names></name><name><surname>Cornelissen</surname><given-names>B</given-names></name><name><surname>Woodcock</surname><given-names>M</given-names></name><name><surname>Allen</surname><given-names>PD</given-names></name><name><surname>Buls</surname><given-names>N</given-names></name><name><surname>Muschel</surname><given-names>RJ</given-names></name><name><surname>Hill</surname><given-names>MA</given-names></name><name><surname>Smart</surname><given-names>SC</given-names></name></person-group> (<year>2011</year>)<article-title> Micro-CT for anatomic referencing in PET and SPECT: radiation dose, biologic damage, and image quality.</article-title><source>The Journal of Nuclear Medicine </source><volume>52</volume>: <fpage>1827</fpage>-<lpage>1833</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.2967/jnumed.111.089151">10.2967/jnumed.111.089151</ext-link></mixed-citation></ref><ref id="B56"><mixed-citation publication-type="other"><person-group><name><surname>Knapp</surname><given-names>S</given-names></name><name><surname>Bateman</surname><given-names>RM</given-names></name><name><surname>Chalmers</surname><given-names>NR</given-names></name><name><surname>Humphries</surname><given-names>CJ</given-names></name><name><surname>Rainbow</surname><given-names>PS</given-names></name><name><surname>Smith</surname><given-names>AB</given-names></name><name><surname>Taylor</surname><given-names>PD</given-names></name><name><surname>Vane-Wright</surname><given-names>RI</given-names></name><name><surname>Wilkinson</surname><given-names>M</given-names></name></person-group> (<year>2002</year>)<source> Taxonomy needs evolution, not revolution. Nature 419: 559. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1038/419559a">10.1038/419559a</ext-link></mixed-citation></ref><ref id="B57"><mixed-citation publication-type="journal"><person-group><name><surname>Kumar</surname><given-names>P</given-names></name><name><surname>Ziegler</surname><given-names>A</given-names></name><name><surname>Grahn</surname><given-names>A</given-names></name><name><surname>Hee</surname><given-names>CS</given-names></name><name><surname>Ziegler</surname><given-names>A</given-names></name></person-group> (<year>2010</year>)<article-title> Leaving the structural ivory tower, assisted by interactive 3D PDF.</article-title><source>Trends in Biochemical Sciences </source><volume>35</volume>: <fpage>419</fpage>-<lpage>422</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.tibs.2010.03.008">10.1016/j.tibs.2010.03.008</ext-link><pub-id pub-id-type="pmid">20541422</pub-id></mixed-citation></ref><pmc-comment>PageBreak</pmc-comment>
<ref id="B58"><mixed-citation publication-type="other"><person-group><name><surname>Laforsch</surname><given-names>C</given-names></name><name><surname>Imhof</surname><given-names>H</given-names></name><name><surname>Sigl</surname><given-names>R</given-names></name><name><surname>Settles</surname><given-names>M</given-names></name><name><surname>Heß</surname><given-names>M</given-names></name><name><surname>Wanninger</surname><given-names>A</given-names></name></person-group> (<year>2012</year>)<article-title> Applications of Computational 3D–Modeling in Organismal Biology.</article-title> In: <person-group><name><surname>Alexandru</surname><given-names>C</given-names></name></person-group> (<role>Ed</role>). <issue-title>Modeling and Simulation in Engineering.</issue-title><source>InTech, Rijeka</source>: <fpage>117</fpage>-<lpage>142</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.5772/31092">10.5772/31092</ext-link></mixed-citation></ref><ref id="B59"><mixed-citation publication-type="other"><person-group><name><surname>Laha</surname><given-names>B</given-names></name><name><surname>Sensharma</surname><given-names>K</given-names></name><name><surname>Schiffbauer</surname><given-names>JD</given-names></name><name><surname>Bowman</surname><given-names>DA</given-names></name></person-group> (<year>2012</year>)<source> Effects of immersion on visual analysis of volume data. IEEE transactions on visualization and computer graphics 18: 597–606. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1109/TVCG.2012.42">10.1109/TVCG.2012.42</ext-link></mixed-citation></ref><ref id="B60"><mixed-citation publication-type="other"><person-group><name><surname>Leuckart</surname><given-names>R</given-names></name></person-group> (<year>1847</year>)<source> Verzeichniss der zur Fauna Helgolands gehörenden wirbellosen Seethiere. In: Frey and Leuckart, Beiträge zur Kenntniss wirbelloser Thiere mit besonderer Berücksichtigung der Fauna des Norddeutschen Meeres. Vieweg, Braunschweig, 136–168. <ext-link ext-link-type="uri" xlink:href="http://www.biodiversitylibrary.org/bibliography/2128">http://www.biodiversitylibrary.org/bibliography/2128</ext-link></source></mixed-citation></ref><ref id="B61"><mixed-citation publication-type="journal"><person-group><name><surname>Marxen</surname><given-names>JC</given-names></name><name><surname>Prymak</surname><given-names>O</given-names></name><name><surname>Beckmann</surname><given-names>F</given-names></name><name><surname>Neues</surname><given-names>F</given-names></name><name><surname>Epple</surname><given-names>M</given-names></name></person-group> (<year>2007</year>)<article-title> Embryonic shell formation in the snail <italic>Biomphalaria glabrata</italic>: a comparison between scanning electron microscopy (SEM) and synchrotron radiation micro computer tomography (SR CT).</article-title><source>Journal of Molluscan Studies </source><volume>74</volume>: <fpage>19</fpage>-<lpage>26</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1093/mollus/eym044">10.1093/mollus/eym044</ext-link></mixed-citation></ref><ref id="B62"><mixed-citation publication-type="journal"><person-group><name><surname>McPeek</surname><given-names>MA</given-names></name><name><surname>Symes</surname><given-names>LB</given-names></name><name><surname>Zong</surname><given-names>DM</given-names></name><name><surname>McPeek</surname><given-names>CL</given-names></name></person-group> (<year>2011</year>)<article-title> Species recognition and patterns of population variation in the reproductive structures of a damselfly genus.</article-title><source>Evolution </source><volume>65</volume>: <fpage>419</fpage>-<lpage>428</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1111/j.1558-5646.2010.01138.x">10.1111/j.1558-5646.2010.01138.x</ext-link><pub-id pub-id-type="pmid">20874736</pub-id></mixed-citation></ref><ref id="B63"><mixed-citation publication-type="other"><person-group><name><surname>Metscher</surname><given-names>BD</given-names></name></person-group> (<year>2009a</year>)<source> MicroCT for comparative morphology: simple staining methods allow high-contrast 3D imaging of diverse non-mineralized animal tissues. BMC Physiology 9: 11. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1186/1472-6793-9-11">10.1186/1472-6793-9-11</ext-link></mixed-citation></ref><ref id="B64"><mixed-citation publication-type="journal"><person-group><name><surname>Metscher</surname><given-names>BD</given-names></name></person-group> (<year>2009b</year>)<article-title> MicroCT for developmental biology: a versatile tool for high-contrast 3D imaging at histological resolutions.</article-title><source>Developmental Dynamics </source><volume>238</volume>: <fpage>632</fpage>-<lpage>640</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1002/dvdy.21857">10.1002/dvdy.21857</ext-link><pub-id pub-id-type="pmid">19235724</pub-id></mixed-citation></ref><ref id="B65"><mixed-citation publication-type="journal"><person-group><name><surname>Midgley</surname><given-names>PA</given-names></name><name><surname>Dunin-Borkowski</surname><given-names>RE</given-names></name></person-group> (<year>2009</year>)<article-title> Electron tomography and holography in materials science.</article-title><source>Nature Materials </source><volume>8</volume>: <fpage>271</fpage>-<lpage>280</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1038/nmat2406">10.1038/nmat2406</ext-link></mixed-citation></ref><ref id="B66"><mixed-citation publication-type="other"><person-group><name><surname>Mildenberger</surname><given-names>P</given-names></name><name><surname>Jensch</surname><given-names>P</given-names></name></person-group> (<year>1999</year>)<source> Implementation of the DICOM standard in a heterogeneous environment: incompatibility or interoperability? Der Radiologe 39: 282–285. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1007/s001170050510">10.1007/s001170050510</ext-link></mixed-citation></ref><ref id="B67"><mixed-citation publication-type="other"><person-group><name><surname>Molineux</surname><given-names>A</given-names></name><name><surname>Scott</surname><given-names>RW</given-names></name><name><surname>Ketcham</surname><given-names>RA</given-names></name><name><surname>Maisano</surname><given-names>JA</given-names></name></person-group> (<year>2007</year>)<source> Rudist taxonomy using X-ray computed tomography. Palaeontologica Electronica 10: 13. <ext-link ext-link-type="uri" xlink:href="http://palaeo-electronica.org/2007_3/135/index.html">http://palaeo-electronica.org/2007_3/135/index.html</ext-link></source></mixed-citation></ref><ref id="B68"><mixed-citation publication-type="journal"><person-group><name><surname>Nickel</surname><given-names>M</given-names></name><name><surname>Donath</surname><given-names>T</given-names></name><name><surname>Schweikert</surname><given-names>M</given-names></name><name><surname>Beckmann</surname><given-names>F</given-names></name></person-group> (<year>2006</year>)<article-title> Functional morphology of <italic>Tethya</italic> species (Porifera): 1. Quantitative 3D-analysis of <italic>Tethya wilhelma</italic> by synchrotron radiation based X-ray microtomography.</article-title><source>Zoomorphology </source><volume>125</volume>: <fpage>209</fpage>-<lpage>223</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1007/s00435-006-0021-1">10.1007/s00435-006-0021-1</ext-link></mixed-citation></ref><ref id="B69"><mixed-citation publication-type="journal"><person-group><name><surname>Nygren</surname><given-names>A</given-names></name><name><surname>Eklöf</surname><given-names>J</given-names></name><name><surname>Pleijel</surname><given-names>F</given-names></name></person-group> (<year>2010</year>)<article-title> Cryptic species of <italic>Notophyllum</italic> (Polychaeta: Phyllodocidae) in Scandinavian waters.</article-title><source>Organisms Diversity & Evolution </source><volume>10</volume>: <fpage>193</fpage>-<lpage>204</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1007/s13127-010-0014-2">10.1007/s13127-010-0014-2</ext-link></mixed-citation></ref><ref id="B70"><mixed-citation publication-type="journal"><person-group><name><surname>Nygren</surname><given-names>A</given-names></name><name><surname>Pleijel</surname><given-names>F</given-names></name></person-group> (<year>2011</year>)<article-title> From one to ten in a single stroke – resolving the European <italic>Eumida sanguinea</italic> (Phyllodocidae, Annelida) species complex.</article-title><source>Molecular phylogenetics and evolution </source><volume>58</volume>: <fpage>132</fpage>-<lpage>141</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.ympev.2010.10.010">10.1016/j.ympev.2010.10.010</ext-link><pub-id pub-id-type="pmid">21034835</pub-id></mixed-citation></ref><ref id="B71"><mixed-citation publication-type="journal"><person-group><name><surname>Osborn</surname><given-names>KJ</given-names></name><name><surname>Rouse</surname><given-names>GW</given-names></name></person-group> (<year>2010</year>)<article-title> Phylogenetics of Acrocirridae and Flabelligeridae (Cirratuliformia, Annelida).</article-title><source>Zoologica Scripta </source><volume>40</volume>: <fpage>204</fpage>-<lpage>219</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1111/j.1463-6409.2010.00460.x">10.1111/j.1463-6409.2010.00460.x</ext-link></mixed-citation></ref><pmc-comment>PageBreak</pmc-comment>
<ref id="B72"><mixed-citation publication-type="other"><person-group><name><surname>Pallas</surname><given-names>PS</given-names></name></person-group> (<year>1766</year>)<source> Miscellanea zoologica. Quibus novae imprimis atque obscurae animalium species describuntur et observationibus iconibusque illustrantur. Petrum van Cleef, Hagí Comitum, 244 pp. <ext-link ext-link-type="uri" xlink:href="http://books.google.gr/books?id=cTQVAAAAQAAJ">http://books.google.gr/books?id=cTQVAAAAQAAJ</ext-link></source></mixed-citation></ref><ref id="B73"><mixed-citation publication-type="journal"><person-group><name><surname>Paredes</surname><given-names>UM</given-names></name><name><surname>Prys-Jones</surname><given-names>R</given-names></name><name><surname>Adams</surname><given-names>M</given-names></name><name><surname>Groombridge</surname><given-names>J</given-names></name><name><surname>Kundu</surname><given-names>S</given-names></name><name><surname>Agapow</surname><given-names>P-M</given-names></name><name><surname>Abel</surname><given-names>RL</given-names></name></person-group> (<year>2012</year>)<article-title> Micro-CT X-rays do not fragment DNA in preserved bird skins.</article-title><source>Journal of Zoological Systematics and Evolutionary Research </source><volume>50</volume>: <fpage>247</fpage>-<lpage>250</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1111/j.1439-0469.2012.00657.x">10.1111/j.1439-0469.2012.00657.x</ext-link></mixed-citation></ref><ref id="B74"><mixed-citation publication-type="journal"><person-group><name><surname>Patek</surname><given-names>SN</given-names></name><name><surname>Nowroozi</surname><given-names>BN</given-names></name><name><surname>Baio</surname><given-names>JE</given-names></name><name><surname>Caldwell</surname><given-names>RL</given-names></name><name><surname>Summers</surname><given-names>AP</given-names></name></person-group> (<year>2007</year>)<article-title> Linkage mechanics and power amplification of the mantis shrimp’s strike.</article-title><source>The Journal of Experimental Biology </source><volume>210</volume>: <fpage>3677</fpage>-<lpage>3688</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1242/jeb.006486">10.1242/jeb.006486</ext-link><pub-id pub-id-type="pmid">17921168</pub-id></mixed-citation></ref><ref id="B75"><mixed-citation publication-type="journal"><person-group><name><surname>Penev</surname><given-names>L</given-names></name><name><surname>Agosti</surname><given-names>D</given-names></name><name><surname>Georgiev</surname><given-names>T</given-names></name><name><surname>Catapano</surname><given-names>T</given-names></name><name><surname>Miller</surname><given-names>J</given-names></name><name><surname>Blagoderov</surname><given-names>V</given-names></name><name><surname>Roberts</surname><given-names>D</given-names></name><name><surname>Smith</surname><given-names>VS</given-names></name><name><surname>Brake</surname><given-names>I</given-names></name><name><surname>Ryrcroft</surname><given-names>S</given-names></name><name><surname>Scott</surname><given-names>B</given-names></name><name><surname>Johnson</surname><given-names>NF</given-names></name><name><surname>Morris</surname><given-names>R a.</given-names></name><name><surname>Sautter</surname><given-names>G</given-names></name><name><surname>Chavan</surname><given-names>V</given-names></name><name><surname>Robertson</surname><given-names>T</given-names></name><name><surname>Remsen</surname><given-names>D</given-names></name><name><surname>Stoev</surname><given-names>P</given-names></name><name><surname>Parr</surname><given-names>C</given-names></name><name><surname>Knapp</surname><given-names>S</given-names></name><name><surname>Kress</surname><given-names>WJ</given-names></name><name><surname>Thompson</surname><given-names>FC</given-names></name><name><surname>Erwin</surname><given-names>T</given-names></name></person-group> (<year>2010</year>)<article-title> Semantic tagging of and semantic enhancements to systematics papers: ZooKeys working examples.</article-title><source>ZooKeys </source><volume>50</volume>: <fpage>1</fpage>-<lpage>16</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.3897/zookeys.50.538">10.3897/zookeys.50.538</ext-link><pub-id pub-id-type="pmid">21594113</pub-id></mixed-citation></ref><ref id="B76"><mixed-citation publication-type="other"><person-group><name><surname>Penney</surname><given-names>D</given-names></name><name><surname>Dierick</surname><given-names>M</given-names></name><name><surname>Cnudde</surname><given-names>V</given-names></name><name><surname>Masschaele</surname><given-names>B</given-names></name><name><surname>Vlassenbroeck</surname><given-names>J</given-names></name><name><surname>Van Hoorebeke,</surname><given-names>L</given-names></name><name><surname>Jacobs</surname><given-names>P</given-names></name></person-group> (<year>2007</year>)<source> First fossil Micropholcommatidae (Araneae), imaged in Eocene Paris amber using X-Ray Computed Tomography. Zootaxa 53: 47–53. <ext-link ext-link-type="uri" xlink:href="https://www.scopus.com/record/display.url?eid=2-s2.0-35748974880&origin=resultslist">https://www.scopus.com/record/display.url?eid=2-s2.0-35748974880&origin=resultslist</ext-link></source></mixed-citation></ref><ref id="B77"><mixed-citation publication-type="journal"><person-group><name><surname>Philippe</surname><given-names>H</given-names></name><name><surname>Derelle</surname><given-names>R</given-names></name><name><surname>Lopez</surname><given-names>P</given-names></name><name><surname>Pick</surname><given-names>K</given-names></name><name><surname>Borchiellini</surname><given-names>C</given-names></name><name><surname>Boury-Esnault</surname><given-names>N</given-names></name><name><surname>Vacelet</surname><given-names>J</given-names></name><name><surname>Renard</surname><given-names>E</given-names></name><name><surname>Houliston</surname><given-names>E</given-names></name><name><surname>Quéinnec</surname><given-names>E</given-names></name><name><surname>Da</surname><given-names>Silva C</given-names></name><name><surname>Wincker</surname><given-names>P</given-names></name><name><surname>Le Guyader</surname><given-names>H</given-names></name><name><surname>Leys</surname><given-names>S</given-names></name><name><surname>Jackson</surname><given-names>DJ</given-names></name><name><surname>Schreiber</surname><given-names>F</given-names></name><name><surname>Erpenbeck</surname><given-names>D</given-names></name><name><surname>Morgenstern</surname><given-names>B</given-names></name><name><surname>Wörheide</surname><given-names>G</given-names></name><name><surname>Manuel</surname><given-names>M</given-names></name></person-group> (<year>2009</year>)<article-title> Phylogenomics revives traditional views on deep animal relationships.</article-title><source>Current Biology </source><volume>19</volume>: <fpage>706</fpage>-<lpage>712</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.cub.2009.02.052">10.1016/j.cub.2009.02.052</ext-link><pub-id pub-id-type="pmid">19345102</pub-id></mixed-citation></ref><ref id="B78"><mixed-citation publication-type="book"><person-group><name><surname>Pianykh</surname><given-names>OS</given-names></name></person-group> (<year>2012</year>)<source> Digital Imaging and Communications in Medicine (DICOM): A Practical Introduction and Survival Guide.</source><publisher-name>Springer</publisher-name>, <publisher-loc>Berlin</publisher-loc>, <lpage>417</lpage> pp.doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1007/978-3-642-10850-1">10.1007/978-3-642-10850-1</ext-link></mixed-citation></ref><ref id="B79"><mixed-citation publication-type="journal"><person-group><name><surname>Postnov</surname><given-names>A</given-names></name><name><surname>de Clerck</surname><given-names>N</given-names></name><name><surname>Sasov</surname><given-names>A</given-names></name><name><surname>Van Dyck</surname><given-names>D</given-names></name></person-group> (<year>2002</year>)<article-title> 3D in-vivo X-ray microtomography of living snails.</article-title><source>Journal of Microscopy </source><volume>205</volume>: <fpage>201</fpage>-<lpage>204</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1046/j.0022-2720.2001.00986.x">10.1046/j.0022-2720.2001.00986.x</ext-link><pub-id pub-id-type="pmid">11879434</pub-id></mixed-citation></ref><ref id="B80"><mixed-citation publication-type="other"><person-group><name><surname>Prohaska</surname><given-names>S</given-names></name><name><surname>Hutanu</surname><given-names>A</given-names></name><name><surname>Hege</surname><given-names>H-C</given-names></name></person-group> (<year>2004</year>)<source> Interactive Exploration of Large Remote Micro-CT Scans. In: Proceedings of the Conference on Visualization ‘04, San Diego (USA), October 2001 IEEE Computer Society Washington, DC, 345–352. <ext-link ext-link-type="uri" xlink:href="https://www.scopus.com/record/display.url?eid=2-s2.0-17044418003&origin=resultslist">https://www.scopus.com/record/display.url?eid=2-s2.0-17044418003&origin=resultslist</ext-link></source></mixed-citation></ref><ref id="B81"><mixed-citation publication-type="journal"><person-group><name><surname>Puce</surname><given-names>S</given-names></name><name><surname>Pica</surname><given-names>D</given-names></name><name><surname>Brun</surname><given-names>F</given-names></name><name><surname>Mancini</surname><given-names>L</given-names></name><name><surname>Bavestrello</surname><given-names>G</given-names></name></person-group> (<year>2012</year>)<article-title> Genus <italic>Distichopora</italic> (Cnidaria, Hydrozoa): from primary cyclosystem to adult pore organisation.</article-title><source>Coral Reefs </source><volume>31</volume>: <fpage>715</fpage>-<lpage>730</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1007/s00338-012-0885-0">10.1007/s00338-012-0885-0</ext-link></mixed-citation></ref><ref id="B82"><mixed-citation publication-type="journal"><person-group><name><surname>Qiong</surname><given-names>X</given-names></name><name><surname>Hengyong</surname><given-names>Y</given-names></name><name><surname>Bennet</surname><given-names>J</given-names></name><name><surname>Peng</surname><given-names>H</given-names></name><name><surname>Zainon</surname><given-names>R</given-names></name><name><surname>Doesburg</surname><given-names>R</given-names></name><name><surname>OpieE</surname><given-names>A</given-names></name><name><surname>Walsh</surname><given-names>M</given-names></name><name><surname>Haiou</surname><given-names>S</given-names></name><name><surname>Butler</surname><given-names>A</given-names></name><name><surname>Butler</surname><given-names>P</given-names></name><name><surname>Xuanqin</surname><given-names>M</given-names></name><name><surname>Ge</surname><given-names>W</given-names></name></person-group> (<year>2012</year>)<article-title> Image Reconstruction for Hybrid True-Color Micro-CT.</article-title><source>IEEE transactions on biomedical engineering </source><volume>59</volume>: <fpage>1711</fpage>-<lpage>1719</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1109/TBME.2012.2192119">10.1109/TBME.2012.2192119</ext-link><pub-id pub-id-type="pmid">22481806</pub-id></mixed-citation></ref><ref id="B83"><mixed-citation publication-type="book"><person-group><name><surname>Quatrefages</surname><given-names>A</given-names></name></person-group> (<year>1866</year>)<source> Histoire naturelle des Annelés marins et d’eau douce. Annélides et Géphyriens, Volume 1.</source><publisher-name>Librarie Encyclopédique de Roret</publisher-name>, <publisher-loc>Paris</publisher-loc>, <lpage>588</lpage> pp.</mixed-citation></ref><pmc-comment>PageBreak</pmc-comment>
<ref id="B84"><mixed-citation publication-type="journal"><person-group><name><surname>Quintarelli</surname><given-names>G</given-names></name><name><surname>Bellocci</surname><given-names>M</given-names></name><name><surname>Geremia</surname><given-names>R</given-names></name></person-group> (<year>1973</year>)<article-title> On phosphotungstic acid staining. IV. Selectivity of the staining reaction.</article-title><source>Journal of Histochemistry & Cytochemistry </source><volume>21</volume>: <fpage>155</fpage>-<lpage>160</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1177/21.2.155">10.1177/21.2.155</ext-link><pub-id pub-id-type="pmid">4121049</pub-id></mixed-citation></ref><ref id="B85"><mixed-citation publication-type="journal"><person-group><name><surname>Rowe</surname><given-names>T</given-names></name><name><surname>Frank</surname><given-names>LR</given-names></name></person-group> (<year>2011</year>)<article-title> The disappearing third dimension.</article-title><source>Science </source><volume>331</volume>: <fpage>712</fpage>-<lpage>714</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1126/science.1202828">10.1126/science.1202828</ext-link><pub-id pub-id-type="pmid">21311010</pub-id></mixed-citation></ref><ref id="B86"><mixed-citation publication-type="other"><person-group><name><surname>Ruthensteiner</surname><given-names>B</given-names></name><name><surname>Baeumler</surname><given-names>N</given-names></name><name><surname>Barnes</surname><given-names>DG</given-names></name></person-group> (<year>2010</year>)<source> Interactive 3D volume rendering in biomedical publications. Micron 41: 886.e1-886.e17. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.micron.2010.03.010">10.1016/j.micron.2010.03.010</ext-link></mixed-citation></ref><ref id="B87"><mixed-citation publication-type="journal"><person-group><name><surname>Ruthensteiner</surname><given-names>B</given-names></name><name><surname>Hess</surname><given-names>M</given-names></name></person-group> (<year>2008</year>)<article-title> Embedding 3D models of biological specimens in PDF publications.</article-title><source>Microscopy Research and Technique </source><volume>71</volume>: <fpage>778</fpage>-<lpage>786</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1002/jemt.20618">10.1002/jemt.20618</ext-link><pub-id pub-id-type="pmid">18785246</pub-id></mixed-citation></ref><ref id="B88"><mixed-citation publication-type="book"><person-group><name><surname>San Martín</surname><given-names>G</given-names></name></person-group> (<year>2003</year>)<source> Annelida Polychaeta II. Syllidae. In: Ramos MA et al. (Eds) Fauna Ibérica, vol. 21.</source><publisher-name>Museo Nacional de Ciencias Naturales, CSIC</publisher-name>, <publisher-loc>Madrid</publisher-loc>, <lpage>554</lpage> pp.</mixed-citation></ref><ref id="B89"><mixed-citation publication-type="journal"><person-group><name><surname>Smith</surname><given-names>V</given-names></name><name><surname>Blagoderov</surname><given-names>V</given-names></name></person-group> (<year>2012</year>)<article-title> Bringing collections out of the dark.</article-title><source>ZooKeys </source><volume>209</volume>: <fpage>1</fpage>-<lpage>6</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.3897/zookeys.209.3699">10.3897/zookeys.209.3699</ext-link><pub-id pub-id-type="pmid">22859874</pub-id></mixed-citation></ref><ref id="B90"><mixed-citation publication-type="journal"><person-group><name><surname>Sutton</surname><given-names>MD</given-names></name></person-group> (<year>2008</year>)<article-title> Tomographic techniques for the study of exceptionally preserved fossils.</article-title><source>Proceedings of the Royal Society of London, Series B </source><volume>275</volume>: <fpage>1587</fpage>-<lpage>1593</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1098/rspb.2008.0263">10.1098/rspb.2008.0263</ext-link><pub-id pub-id-type="pmid">18426749</pub-id></mixed-citation></ref><ref id="B91"><mixed-citation publication-type="other"><person-group><name><surname>Sutton</surname><given-names>MD</given-names></name><name><surname>Garwood</surname><given-names>RJ</given-names></name><name><surname>Siveter</surname><given-names>DJ</given-names></name><name><surname>Siveter</surname><given-names>DJ</given-names></name></person-group> (<year>2012</year>)<source> SPIERS and VAXML; A software toolkit for tomographic visualisation and a format for virtual specimen interchange. Paleontologica electronica 15: 1–15. <ext-link ext-link-type="uri" xlink:href="http://palaeo-electronica.org/content/issue-2-2012-technical-articles/226-virtual-palaeontology-toolkit">http://palaeo-electronica.org/content/issue-2-2012-technical-articles/226-virtual-palaeontology-toolkit</ext-link></source></mixed-citation></ref><ref id="B92"><mixed-citation publication-type="journal"><person-group><name><surname>Tamura</surname><given-names>K</given-names></name><name><surname>Peterson</surname><given-names>D</given-names></name><name><surname>Peterson</surname><given-names>N</given-names></name><name><surname>Stecher</surname><given-names>G</given-names></name><name><surname>Nei</surname><given-names>M</given-names></name><name><surname>Kumar</surname><given-names>S</given-names></name></person-group> (<year>2011</year>)<article-title> MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods.</article-title><source>Molecular Biology and Evolution </source><volume>28</volume>: <fpage>2731</fpage>-<lpage>2739</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1093/molbev/msr121">10.1093/molbev/msr121</ext-link><pub-id pub-id-type="pmid">21546353</pub-id></mixed-citation></ref><ref id="B93"><mixed-citation publication-type="journal"><person-group><name><surname>Wang</surname><given-names>G</given-names></name><name><surname>Yu</surname><given-names>H</given-names></name><name><surname>De Man</surname><given-names>B</given-names></name></person-group> (<year>2008</year>)<article-title> An outlook on x-ray CT research and development.</article-title><source>Medical Physics </source><volume>35</volume>: <fpage>1051</fpage>-<lpage>1064</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1118/1.2836950">10.1118/1.2836950</ext-link><pub-id pub-id-type="pmid">18404940</pub-id></mixed-citation></ref><ref id="B94"><mixed-citation publication-type="journal"><person-group><name><surname>Westheide</surname><given-names>W</given-names></name></person-group> (<year>1997</year>)<article-title> The direction of evolution within the Polychaeta.</article-title><source>Journal of Natural History </source><volume>31</volume>: <fpage>1</fpage>-<lpage>15</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1080/00222939700770011">10.1080/00222939700770011</ext-link></mixed-citation></ref><ref id="B95"><mixed-citation publication-type="other"><person-group><name><surname>Wheeler</surname><given-names>QD</given-names></name><name><surname>Raven</surname><given-names>PH</given-names></name><name><surname>Wilson</surname><given-names>EO</given-names></name></person-group> (<year>2004</year>)<source> Taxonomy: impediment or expedient? Science 303: 285. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1126/science.303.5656.285">10.1126/science.303.5656.285</ext-link></mixed-citation></ref><ref id="B96"><mixed-citation publication-type="journal"><person-group><name><surname>Wheeler</surname><given-names>Q</given-names></name><name><surname>Thierry</surname><given-names>B</given-names></name><name><surname>Coddington</surname><given-names>J</given-names></name><name><surname>Gostony</surname><given-names>T</given-names></name><name><surname>Hamilton</surname><given-names>A</given-names></name><name><surname>Larimer</surname><given-names>R</given-names></name><name><surname>Polaszek</surname><given-names>A</given-names></name><name><surname>Schauff</surname><given-names>M</given-names></name><name><surname>Solis</surname><given-names>A</given-names></name></person-group> (<year>2012</year>)<article-title> Nomenclatural benchmarking: the roles of digital typification and telemicroscopy.</article-title><source>ZooKeys </source><volume>209</volume>: <fpage>193</fpage>-<lpage>202</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.3897/zookeys.209.3486">10.3897/zookeys.209.3486</ext-link><pub-id pub-id-type="pmid">22859888</pub-id></mixed-citation></ref><ref id="B97"><mixed-citation publication-type="journal"><person-group><name><surname>Wilhelm</surname><given-names>G</given-names></name><name><surname>Handschuh</surname><given-names>S</given-names></name><name><surname>Plant</surname><given-names>J</given-names></name><name><surname>Nemeschkal</surname><given-names>HL</given-names></name></person-group> (<year>2011</year>)<article-title> Sexual dimorphism in head structures of the weevil <italic>Rhopalapion longirostre</italic> (Olivier 1807) (Coleoptera: Curculionoidea): a response to ecological demands of egg deposition.</article-title><source>Biological Journal of the Linnean Society </source><volume>104</volume>: <fpage>642</fpage>-<lpage>660</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1111/j.1095-8312.2011.01751.x">10.1111/j.1095-8312.2011.01751.x</ext-link></mixed-citation></ref><ref id="B98"><mixed-citation publication-type="journal"><person-group><name><surname>Wirkner</surname><given-names>CS</given-names></name><name><surname>Prendini</surname><given-names>L</given-names></name></person-group> (<year>2007</year>)<article-title> Comparative Morphology of the Hemolymph Vascular System in Scorpions–A Survey Using Corrosion Casting, MicroCT, and 3D-Reconstruction.</article-title><source>Journal of Morphology </source><volume>413</volume>: <fpage>401</fpage>-<lpage>413</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1002/jmor.10512">10.1002/jmor.10512</ext-link><pub-id pub-id-type="pmid">17372915</pub-id></mixed-citation></ref><ref id="B99"><mixed-citation publication-type="journal"><person-group><name><surname>Wolff</surname><given-names>S</given-names></name></person-group> (<year>1967</year>)<article-title> Radiation Genetics.</article-title><source>Annual Review of Genetics </source><volume>1</volume>: <fpage>221</fpage>-<lpage>244</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1146/annurev.ge.01.120167.001253">10.1146/annurev.ge.01.120167.001253</ext-link></mixed-citation></ref><pmc-comment>PageBreak</pmc-comment>
<ref id="B100"><mixed-citation publication-type="journal"><person-group><name><surname>Zanol</surname><given-names>J</given-names></name><name><surname>Bettoso</surname><given-names>N</given-names></name></person-group> (<year>2006</year>)<article-title> Identity of <italic>Eunice roussaei</italic> (Eunicidae: Polychaeta: Annelida) from the Adriatic and Mediterranean Seas.</article-title><source>Journal of the Marine Biological Association of the United Kingdom </source><volume>86</volume>: <fpage>1017</fpage>-<lpage>1024</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1017/S0025315406013993">10.1017/S0025315406013993</ext-link></mixed-citation></ref><ref id="B101"><mixed-citation publication-type="journal"><person-group><name><surname>Zanol</surname><given-names>J</given-names></name><name><surname>Halanych</surname><given-names>KM</given-names></name><name><surname>Struck</surname><given-names>TH</given-names></name><name><surname>Fauchald</surname><given-names>K</given-names></name></person-group> (<year>2010</year>)<article-title> Phylogeny of the bristle worm family Eunicidae (Eunicida, Annelida) and the phylogenetic utility of noncongruent 16S, COI and 18S in combined analyses.</article-title><source>Molecular Phylogenetics and Evolution </source><volume>55</volume>: <fpage>660</fpage>-<lpage>676</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.ympev.2009.12.024">10.1016/j.ympev.2009.12.024</ext-link><pub-id pub-id-type="pmid">20040377</pub-id></mixed-citation></ref><ref id="B102"><mixed-citation publication-type="other"><person-group><name><surname>Ziegler</surname><given-names>A</given-names></name><name><surname>Faber</surname><given-names>C</given-names></name><name><surname>Mueller</surname><given-names>S</given-names></name><name><surname>Bartolomaeus</surname><given-names>T</given-names></name></person-group> (<year>2008</year>)<source> Systematic comparison and reconstruction of sea urchin (Echinoidea) internal anatomy: a novel approach using magnetic resonance imaging. BMC Biology 6: 33. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1186/1741-7007-6-33">10.1186/1741-7007-6-33</ext-link></mixed-citation></ref><ref id="B103"><mixed-citation publication-type="journal"><person-group><name><surname>Ziegler</surname><given-names>A</given-names></name><name><surname>Kunth</surname><given-names>M</given-names></name><name><surname>Mueller</surname><given-names>S</given-names></name><name><surname>Bock</surname><given-names>C</given-names></name><name><surname>Pohmann</surname><given-names>R</given-names></name><name><surname>Schröder</surname><given-names>L</given-names></name><name><surname>Faber</surname><given-names>C</given-names></name><name><surname>Giribet</surname><given-names>G</given-names></name></person-group> (<year>2011a</year>)<article-title> Application of magnetic resonance imaging in zoology.</article-title><source>Zoomorphology </source><volume>130</volume>: <fpage>227</fpage>-<lpage>254</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1007/s00435-011-0138-8">10.1007/s00435-011-0138-8</ext-link></mixed-citation></ref><ref id="B104"><mixed-citation publication-type="other"><person-group><name><surname>Ziegler</surname><given-names>A</given-names></name><name><surname>Mietchen</surname><given-names>D</given-names></name><name><surname>Faber</surname><given-names>C</given-names></name><name><surname>von Hausen</surname><given-names>W</given-names></name><name><surname>Schöbel</surname><given-names>C</given-names></name><name><surname>Sellerer</surname><given-names>M</given-names></name><name><surname>Ziegler</surname><given-names>A</given-names></name></person-group> (<year>2011b</year>)<source> Effectively incorporating selected multimedia content into medical publications. BMC Medicine 9: 17. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1186/1741-7015-9-17">10.1186/1741-7015-9-17</ext-link></mixed-citation></ref><ref id="B105"><mixed-citation publication-type="other"><person-group><name><surname>Ziegler</surname><given-names>A</given-names></name><name><surname>Ogurreck</surname><given-names>M</given-names></name><name><surname>Steinke</surname><given-names>T</given-names></name><name><surname>Beckmann</surname><given-names>F</given-names></name><name><surname>Prohaska</surname><given-names>S</given-names></name><name><surname>Ziegler</surname><given-names>A</given-names></name></person-group> (<year>2010</year>)<source> Opportunities and challenges for digital morphology. Biology Direct 5: 45. </source>doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1186/1745-6150-5-45">10.1186/1745-6150-5-45</ext-link></mixed-citation></ref><ref id="B106"><mixed-citation publication-type="journal"><person-group><name><surname>Zimmermann</surname><given-names>D</given-names></name><name><surname>Randolf</surname><given-names>S</given-names></name><name><surname>Metscher</surname><given-names>BD</given-names></name><name><surname>Aspöck</surname><given-names>U</given-names></name></person-group> (<year>2011</year>)<article-title> The function and phylogenetic implications of the tentorium in adult Neuroptera (Insecta).</article-title><source>Arthropod Structure & Development </source><volume>40</volume>: <fpage>571</fpage>-<lpage>582</lpage> doi: <ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1016/j.asd.2011.06.003">10.1016/j.asd.2011.06.003</ext-link><pub-id pub-id-type="pmid">21978822</pub-id></mixed-citation></ref></ref-list></back></pmc></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Belgique/explor/OpenAccessBelV2/Data/Pmc/Corpus
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000442 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Pmc/Corpus/biblio.hfd -nk 000442 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Wicri/Belgique
|area= OpenAccessBelV2
|flux= Pmc
|étape= Corpus
|type= RBID
|clé= PMC:3591762
|texte= Micro-computed tomography: Introducing new dimensions to taxonomy
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Pmc/Corpus/RBID.i -Sk "pubmed:23653515" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Corpus/biblio.hfd \
| NlmPubMed2Wicri -a OpenAccessBelV2
| This area was generated with Dilib version V0.6.25. |  |