Laser Capture Microdissection Protocol for Xylem Tissues of Woody Plants.
Identifieur interne : 001796 ( Main/Exploration ); précédent : 001795; suivant : 001797Laser Capture Microdissection Protocol for Xylem Tissues of Woody Plants.
Auteurs : Olga Blokhina [Finlande] ; Concetta Valerio [Suède] ; Katarzyna Sokołowska [Pologne] ; Lei Zhao [Finlande] ; Anna K Rkönen [Finlande] ; Totte Niittyl [Suède] ; Kurt Fagerstedt [Finlande]Source :
- Frontiers in plant science [ 1664-462X ] ; 2016.
Abstract
Laser capture microdissection (LCM) enables precise dissection and collection of individual cell types from complex tissues. When applied to plant cells, and especially to woody tissues, LCM requires extensive optimization to overcome such factors as rigid cell walls, large central vacuoles, intercellular spaces, and technical issues with thickness and flatness of the sections. Here we present an optimized protocol for the laser-assisted microdissection of developing xylem from mature trees: a gymnosperm (Norway spruce, Picea abies) and an angiosperm (aspen, Populus tremula) tree. Different cell types of spruce and aspen wood (i.e., ray cells, tracheary elements, and fibers) were successfully microdissected from tangential, cross and radial cryosections of the current year's growth ring. Two approaches were applied to achieve satisfactory flatness and anatomical integrity of the spruce and aspen specimens. The commonly used membrane slides were ineffective as a mounting surface for the wood cryosections. Instead, in the present protocol we use glass slides, and introduce a glass slide sandwich assembly for the preparation of aspen sections. To ascertain that not only the anatomical integrity of the plant tissue, but also the molecular features were not compromised during the whole LCM procedure, good quality total RNA could be extracted from the microdissected cells. This showed the efficiency of the protocol and established that our methodology can be integrated in transcriptome analyses to elucidate cell-specific molecular events regulating wood formation in trees.
DOI: 10.3389/fpls.2016.01965
PubMed: 28101088
PubMed Central: PMC5209384
Affiliations:
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Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural SciencesUmeå, Sweden.</nlm:affiliation><country xml:lang="fr">Suède</country><wicri:regionArea>Plant Stress Signaling, Instituto Gulbenkian de CiênciaOeiras, Portugal; Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural SciencesUmeå</wicri:regionArea><wicri:noRegion>Swedish University of Agricultural SciencesUmeå</wicri:noRegion></affiliation></author><author><name sortKey="Sokolowska, Katarzyna" sort="Sokolowska, Katarzyna" uniqKey="Sokolowska K" first="Katarzyna" last="Sokołowska">Katarzyna Sokołowska</name><affiliation wicri:level="1"><nlm:affiliation>Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural SciencesUmeå, Sweden; Department of Plant Developmental Biology, Institute of Experimental Biology, University of WrocławWrocław, Poland.</nlm:affiliation><country xml:lang="fr">Pologne</country><wicri:regionArea>Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural SciencesUmeå, Sweden; Department of Plant Developmental Biology, Institute of Experimental Biology, University of WrocławWrocław</wicri:regionArea><wicri:noRegion>University of WrocławWrocław</wicri:noRegion></affiliation></author><author><name sortKey="Zhao, Lei" sort="Zhao, Lei" uniqKey="Zhao L" first="Lei" last="Zhao">Lei Zhao</name><affiliation wicri:level="1"><nlm:affiliation>Viikki Plant Science Centre, Department of Biosciences, University of Helsinki Helsinki, Finland.</nlm:affiliation><country xml:lang="fr">Finlande</country><wicri:regionArea>Viikki Plant Science Centre, Department of Biosciences, University of Helsinki Helsinki</wicri:regionArea><wicri:noRegion>University of Helsinki Helsinki</wicri:noRegion></affiliation></author><author><name sortKey="K Rkonen, Anna" sort="K Rkonen, Anna" uniqKey="K Rkonen A" first="Anna" last="K Rkönen">Anna K Rkönen</name><affiliation wicri:level="1"><nlm:affiliation>Viikki Plant Science Centre, Department of Agricultural Sciences, University of Helsinki Helsinki, Finland.</nlm:affiliation><country xml:lang="fr">Finlande</country><wicri:regionArea>Viikki Plant Science Centre, Department of Agricultural Sciences, University of Helsinki Helsinki</wicri:regionArea><wicri:noRegion>University of Helsinki Helsinki</wicri:noRegion></affiliation></author><author><name sortKey="Niittyl, Totte" sort="Niittyl, Totte" uniqKey="Niittyl T" first="Totte" last="Niittyl">Totte Niittyl</name><affiliation wicri:level="1"><nlm:affiliation>Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences Umeå, Sweden.</nlm:affiliation><country xml:lang="fr">Suède</country><wicri:regionArea>Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences Umeå</wicri:regionArea><wicri:noRegion>Swedish University of Agricultural Sciences Umeå</wicri:noRegion></affiliation></author><author><name sortKey="Fagerstedt, Kurt" sort="Fagerstedt, Kurt" uniqKey="Fagerstedt K" first="Kurt" last="Fagerstedt">Kurt Fagerstedt</name><affiliation wicri:level="1"><nlm:affiliation>Viikki Plant Science Centre, Department of Biosciences, University of Helsinki Helsinki, Finland.</nlm:affiliation><country xml:lang="fr">Finlande</country><wicri:regionArea>Viikki Plant Science Centre, Department of Biosciences, University of Helsinki Helsinki</wicri:regionArea><wicri:noRegion>University of Helsinki Helsinki</wicri:noRegion></affiliation></author></analytic><series><title level="j">Frontiers in plant science</title><idno type="ISSN">1664-462X</idno><imprint><date when="2016" type="published">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Laser capture microdissection (LCM) enables precise dissection and collection of individual cell types from complex tissues. When applied to plant cells, and especially to woody tissues, LCM requires extensive optimization to overcome such factors as rigid cell walls, large central vacuoles, intercellular spaces, and technical issues with thickness and flatness of the sections. Here we present an optimized protocol for the laser-assisted microdissection of developing xylem from mature trees: a gymnosperm (Norway spruce, <i>Picea abies</i>) and an angiosperm (aspen, <i>Populus tremula</i>) tree. Different cell types of spruce and aspen wood (i.e., ray cells, tracheary elements, and fibers) were successfully microdissected from tangential, cross and radial cryosections of the current year's growth ring. Two approaches were applied to achieve satisfactory flatness and anatomical integrity of the spruce and aspen specimens. The commonly used membrane slides were ineffective as a mounting surface for the wood cryosections. Instead, in the present protocol we use glass slides, and introduce a glass slide sandwich assembly for the preparation of aspen sections. To ascertain that not only the anatomical integrity of the plant tissue, but also the molecular features were not compromised during the whole LCM procedure, good quality total RNA could be extracted from the microdissected cells. This showed the efficiency of the protocol and established that our methodology can be integrated in transcriptome analyses to elucidate cell-specific molecular events regulating wood formation in trees.</div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">28101088</PMID><DateRevised><Year>2020</Year><Month>10</Month><Day>01</Day></DateRevised><Article PubModel="Electronic-eCollection"><Journal><ISSN IssnType="Print">1664-462X</ISSN><JournalIssue CitedMedium="Print"><Volume>7</Volume><PubDate><Year>2016</Year></PubDate></JournalIssue><Title>Frontiers in plant science</Title><ISOAbbreviation>Front Plant Sci</ISOAbbreviation></Journal><ArticleTitle>Laser Capture Microdissection Protocol for Xylem Tissues of Woody Plants.</ArticleTitle><Pagination><MedlinePgn>1965</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.3389/fpls.2016.01965</ELocationID><Abstract><AbstractText>Laser capture microdissection (LCM) enables precise dissection and collection of individual cell types from complex tissues. When applied to plant cells, and especially to woody tissues, LCM requires extensive optimization to overcome such factors as rigid cell walls, large central vacuoles, intercellular spaces, and technical issues with thickness and flatness of the sections. Here we present an optimized protocol for the laser-assisted microdissection of developing xylem from mature trees: a gymnosperm (Norway spruce, <i>Picea abies</i>) and an angiosperm (aspen, <i>Populus tremula</i>) tree. Different cell types of spruce and aspen wood (i.e., ray cells, tracheary elements, and fibers) were successfully microdissected from tangential, cross and radial cryosections of the current year's growth ring. Two approaches were applied to achieve satisfactory flatness and anatomical integrity of the spruce and aspen specimens. The commonly used membrane slides were ineffective as a mounting surface for the wood cryosections. Instead, in the present protocol we use glass slides, and introduce a glass slide sandwich assembly for the preparation of aspen sections. To ascertain that not only the anatomical integrity of the plant tissue, but also the molecular features were not compromised during the whole LCM procedure, good quality total RNA could be extracted from the microdissected cells. This showed the efficiency of the protocol and established that our methodology can be integrated in transcriptome analyses to elucidate cell-specific molecular events regulating wood formation in trees.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Blokhina</LastName><ForeName>Olga</ForeName><Initials>O</Initials><AffiliationInfo><Affiliation>Viikki Plant Science Centre, Department of Biosciences, University of Helsinki Helsinki, Finland.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Valerio</LastName><ForeName>Concetta</ForeName><Initials>C</Initials><AffiliationInfo><Affiliation>Plant Stress Signaling, Instituto Gulbenkian de CiênciaOeiras, Portugal; Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural SciencesUmeå, Sweden.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Sokołowska</LastName><ForeName>Katarzyna</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural SciencesUmeå, Sweden; Department of Plant Developmental Biology, Institute of Experimental Biology, University of WrocławWrocław, Poland.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhao</LastName><ForeName>Lei</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>Viikki Plant Science Centre, Department of Biosciences, University of Helsinki Helsinki, Finland.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Kärkönen</LastName><ForeName>Anna</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Viikki Plant Science Centre, Department of Agricultural Sciences, University of Helsinki Helsinki, Finland.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Niittylä</LastName><ForeName>Totte</ForeName><Initials>T</Initials><AffiliationInfo><Affiliation>Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences Umeå, Sweden.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Fagerstedt</LastName><ForeName>Kurt</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>Viikki Plant Science Centre, Department of Biosciences, University of Helsinki Helsinki, Finland.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2017</Year><Month>01</Month><Day>04</Day></ArticleDate></Article><MedlineJournalInfo><Country>Switzerland</Country><MedlineTA>Front Plant Sci</MedlineTA><NlmUniqueID>101568200</NlmUniqueID><ISSNLinking>1664-462X</ISSNLinking></MedlineJournalInfo><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">RNA integrity</Keyword><Keyword MajorTopicYN="N">cryosection</Keyword><Keyword MajorTopicYN="N">laser capture microdissection</Keyword><Keyword MajorTopicYN="N">ray cells</Keyword><Keyword MajorTopicYN="N">tracheids</Keyword><Keyword MajorTopicYN="N">xylem fibers</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2016</Year><Month>10</Month><Day>24</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2016</Year><Month>12</Month><Day>12</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2017</Year><Month>1</Month><Day>20</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2017</Year><Month>1</Month><Day>20</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2017</Year><Month>1</Month><Day>20</Day><Hour>6</Hour><Minute>1</Minute></PubMedPubDate></History><PublicationStatus>epublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">28101088</ArticleId><ArticleId IdType="doi">10.3389/fpls.2016.01965</ArticleId><ArticleId IdType="pmc">PMC5209384</ArticleId></ArticleIdList><ReferenceList><Reference><Citation>Plant Physiol. 2003 May;132(1):27-35</Citation><ArticleIdList><ArticleId IdType="pubmed">12746508</ArticleId></ArticleIdList></Reference><Reference><Citation>J Biomed Opt. 2007 Sep-Oct;12(5):054016</Citation><ArticleIdList><ArticleId IdType="pubmed">17994904</ArticleId></ArticleIdList></Reference><Reference><Citation>New Phytol. 2011 Apr;190(2):351-68</Citation><ArticleIdList><ArticleId IdType="pubmed">21091694</ArticleId></ArticleIdList></Reference><Reference><Citation>Plant Physiol. 2016 Jul;171(3):2256-76</Citation><ArticleIdList><ArticleId 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IdType="pubmed">22992511</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed><affiliations><list><country><li>Finlande</li><li>Pologne</li><li>Suède</li></country></list><tree><country name="Finlande"><noRegion><name sortKey="Blokhina, Olga" sort="Blokhina, Olga" uniqKey="Blokhina O" first="Olga" last="Blokhina">Olga Blokhina</name></noRegion><name sortKey="Fagerstedt, Kurt" sort="Fagerstedt, Kurt" uniqKey="Fagerstedt K" first="Kurt" last="Fagerstedt">Kurt Fagerstedt</name><name sortKey="K Rkonen, Anna" sort="K Rkonen, Anna" uniqKey="K Rkonen A" first="Anna" last="K Rkönen">Anna K Rkönen</name><name sortKey="Zhao, Lei" sort="Zhao, Lei" uniqKey="Zhao L" first="Lei" last="Zhao">Lei Zhao</name></country><country name="Suède"><noRegion><name sortKey="Valerio, Concetta" sort="Valerio, Concetta" uniqKey="Valerio C" first="Concetta" last="Valerio">Concetta Valerio</name></noRegion><name sortKey="Niittyl, Totte" sort="Niittyl, Totte" uniqKey="Niittyl T" first="Totte" last="Niittyl">Totte Niittyl</name></country><country name="Pologne"><noRegion><name sortKey="Sokolowska, Katarzyna" sort="Sokolowska, Katarzyna" uniqKey="Sokolowska K" first="Katarzyna" last="Sokołowska">Katarzyna Sokołowska</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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