In situ reverse transcription-polymerase chain reaction. Applications for light and electron microscopy
Identifieur interne : 002518 ( Istex/Corpus ); précédent : 002517; suivant : 002519In situ reverse transcription-polymerase chain reaction. Applications for light and electron microscopy
Auteurs : Gérard Morel ; Martina Berger ; Brice Ronsin ; Sophie Recher ; Sylvie Ricard-Blum ; Hichem C. Mertani ; Peter E. LobieSource :
- Biology of the Cell [ 0248-4900 ] ; 1998.
English descriptors
- Teeft :
- Alkaline phosphatase, Amplification, Amplification mixture, Amplification products, Amplification reaction, Annealing, Appropriate controls, Avian myeloblastosis virus, Bagasra, Biological material, Boca york, Boehringer mannheim, Careful optimization, Cdna, Cell membrane, Cell suspensions, Cycle number, Denaturation, Detection system, Diffusion artifacts, Digestion, Digestion step, Direct reaction, Dnase, Dnase digestion, Dntps, Electron microscopy, Elongation step, Embedding, Endocrine cell populations, Endogenous biotin, Ethanol, Extension time, Final concentration, First step, Fixation, Fixation time, Fixative, Gene expression, Gene rearrangements, Genome, Glass slides, Gold particles, Good preservation, Growth hormone receptor mrna, Haase, High number, Higher concentration, Host cell, Human papillomavirus, Human press, Humid chamber, Hybridization, Hybridization step, Hybridization temperature, Hydrophylic resin, Indirect reactions, Individual cells, Infectious agents, Komminoth, Light microscopy, Localization, Major pitfall, Mertani, Mispriming, Molecular biology, Molecular diagnosis, Morel, Mrna, Mrna coding, Negative strand, Nucleic, Nucleic acid, Nucleic acid sequences, Nucleic acid target, Nucleic acids, Nucleotide, Nuovo, Paraformaldehyde, Phosphate buffer, Pituitary gland section, Polymerase, Polymerase activity, Polymerase chain reaction, Positive cells, Positive strand, Possible adsorption, Possible degradation, Powerful technique, Primer, Proteolytic digestion, Protocol, Raven press, Receptor, Replication, Rnase, Ronsin, Room temperature, Second step, Secondary structures, Several approaches, Signal density, Target sequence, Tissue preparation, Tissue section, Tissue sections, Total volume, Transcriptase, Ultrathin, Ultrathin sections, Unlabeled nucleotides, Unlabeled probe, Vibratome sections, Viral, Viral genome, Viral methods, Visna virus, White resin.
Abstract
Abstract: Since its discovery in 1986 by Mullis, the polymerase chain reaction (PCR) has been extensively developed by morphologists in order to overcome the main limitation of in situ hybridization, the lack of sensitivity. In situ PCR combines the extreme sensitivity of PCR with the cell-localizing ability of in situ hybridization. The amplification of DNA (PCR) or a cDNA (RT-PCR) in cell or tissue sections has been developed at light and electron microscopic levels. A successful PCR experiment requires the careful optimization of several parameters depending on the tissue (or of cell types), and a compromise must be found between the fixation time, pretreatments and a good preservation of the morphology. Other crucial factors (primer design, concentration in MgCl2, annealing and elongation temperatures during the amplification steps) and their influence on the specificity and sensitivity of in situ PCR or RT-PCR are discussed. The necessity to run appropriate controls, especially to assess the lack of diffusion of the amplified products, is stressed. Current applications and future trends are also presented.
Url:
DOI: 10.1016/S0248-4900(98)80335-3
Links to Exploration step
ISTEX:7DC11D66EC5B5B3B9875983BDAFD1A98F370D651Le document en format XML
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Applications for light and electron microscopy</title><author><name sortKey="Morel, Gerard" sort="Morel, Gerard" uniqKey="Morel G" first="Gérard" last="Morel">Gérard Morel</name><affiliation><mods:affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Berger, Martina" sort="Berger, Martina" uniqKey="Berger M" first="Martina" last="Berger">Martina Berger</name><affiliation><mods:affiliation>Laboratoire de Virologie, Faculté de Médecine, Domaine Rockefeller, Lyon, France</mods:affiliation></affiliation></author><author><name sortKey="Ronsin, Brice" sort="Ronsin, Brice" uniqKey="Ronsin B" first="Brice" last="Ronsin">Brice Ronsin</name><affiliation><mods:affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Recher, Sophie" sort="Recher, Sophie" uniqKey="Recher S" first="Sophie" last="Recher">Sophie Recher</name><affiliation><mods:affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Ricard Blum, Sylvie" sort="Ricard Blum, Sylvie" uniqKey="Ricard Blum S" first="Sylvie" last="Ricard-Blum">Sylvie Ricard-Blum</name><affiliation><mods:affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Mertani, Hichem C" sort="Mertani, Hichem C" uniqKey="Mertani H" first="Hichem C" last="Mertani">Hichem C. 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Applications for light and electron microscopy</title><author><name sortKey="Morel, Gerard" sort="Morel, Gerard" uniqKey="Morel G" first="Gérard" last="Morel">Gérard Morel</name><affiliation><mods:affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Berger, Martina" sort="Berger, Martina" uniqKey="Berger M" first="Martina" last="Berger">Martina Berger</name><affiliation><mods:affiliation>Laboratoire de Virologie, Faculté de Médecine, Domaine Rockefeller, Lyon, France</mods:affiliation></affiliation></author><author><name sortKey="Ronsin, Brice" sort="Ronsin, Brice" uniqKey="Ronsin B" first="Brice" last="Ronsin">Brice Ronsin</name><affiliation><mods:affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Recher, Sophie" sort="Recher, Sophie" uniqKey="Recher S" first="Sophie" last="Recher">Sophie Recher</name><affiliation><mods:affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Ricard Blum, Sylvie" sort="Ricard Blum, Sylvie" uniqKey="Ricard Blum S" first="Sylvie" last="Ricard-Blum">Sylvie Ricard-Blum</name><affiliation><mods:affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Mertani, Hichem C" sort="Mertani, Hichem C" uniqKey="Mertani H" first="Hichem C" last="Mertani">Hichem C. Mertani</name><affiliation><mods:affiliation>Institute of Molecular and Cell Biology, National University of Singapore, Singapore</mods:affiliation></affiliation></author><author><name sortKey="Lobie, Peter E" sort="Lobie, Peter E" uniqKey="Lobie P" first="Peter E" last="Lobie">Peter E. Lobie</name><affiliation><mods:affiliation>Institute of Molecular and Cell Biology, National University of Singapore, Singapore</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Biology of the Cell</title><title level="j" type="abbrev">BICELL</title><idno type="ISSN">0248-4900</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1998">1998</date><biblScope unit="volume">90</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="137">137</biblScope><biblScope unit="page" to="154">154</biblScope></imprint><idno type="ISSN">0248-4900</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0248-4900</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Alkaline phosphatase</term><term>Amplification</term><term>Amplification mixture</term><term>Amplification products</term><term>Amplification reaction</term><term>Annealing</term><term>Appropriate controls</term><term>Avian myeloblastosis virus</term><term>Bagasra</term><term>Biological material</term><term>Boca york</term><term>Boehringer mannheim</term><term>Careful optimization</term><term>Cdna</term><term>Cell membrane</term><term>Cell suspensions</term><term>Cycle number</term><term>Denaturation</term><term>Detection system</term><term>Diffusion artifacts</term><term>Digestion</term><term>Digestion step</term><term>Direct reaction</term><term>Dnase</term><term>Dnase digestion</term><term>Dntps</term><term>Electron microscopy</term><term>Elongation step</term><term>Embedding</term><term>Endocrine cell populations</term><term>Endogenous biotin</term><term>Ethanol</term><term>Extension time</term><term>Final concentration</term><term>First step</term><term>Fixation</term><term>Fixation time</term><term>Fixative</term><term>Gene expression</term><term>Gene rearrangements</term><term>Genome</term><term>Glass slides</term><term>Gold particles</term><term>Good preservation</term><term>Growth hormone receptor mrna</term><term>Haase</term><term>High number</term><term>Higher concentration</term><term>Host cell</term><term>Human papillomavirus</term><term>Human press</term><term>Humid chamber</term><term>Hybridization</term><term>Hybridization step</term><term>Hybridization temperature</term><term>Hydrophylic resin</term><term>Indirect reactions</term><term>Individual cells</term><term>Infectious agents</term><term>Komminoth</term><term>Light microscopy</term><term>Localization</term><term>Major pitfall</term><term>Mertani</term><term>Mispriming</term><term>Molecular biology</term><term>Molecular diagnosis</term><term>Morel</term><term>Mrna</term><term>Mrna coding</term><term>Negative strand</term><term>Nucleic</term><term>Nucleic acid</term><term>Nucleic acid sequences</term><term>Nucleic acid target</term><term>Nucleic acids</term><term>Nucleotide</term><term>Nuovo</term><term>Paraformaldehyde</term><term>Phosphate buffer</term><term>Pituitary gland section</term><term>Polymerase</term><term>Polymerase activity</term><term>Polymerase chain reaction</term><term>Positive cells</term><term>Positive strand</term><term>Possible adsorption</term><term>Possible degradation</term><term>Powerful technique</term><term>Primer</term><term>Proteolytic digestion</term><term>Protocol</term><term>Raven press</term><term>Receptor</term><term>Replication</term><term>Rnase</term><term>Ronsin</term><term>Room temperature</term><term>Second step</term><term>Secondary structures</term><term>Several approaches</term><term>Signal density</term><term>Target sequence</term><term>Tissue preparation</term><term>Tissue section</term><term>Tissue sections</term><term>Total volume</term><term>Transcriptase</term><term>Ultrathin</term><term>Ultrathin sections</term><term>Unlabeled nucleotides</term><term>Unlabeled probe</term><term>Vibratome sections</term><term>Viral</term><term>Viral genome</term><term>Viral methods</term><term>Visna virus</term><term>White resin</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Since its discovery in 1986 by Mullis, the polymerase chain reaction (PCR) has been extensively developed by morphologists in order to overcome the main limitation of in situ hybridization, the lack of sensitivity. In situ PCR combines the extreme sensitivity of PCR with the cell-localizing ability of in situ hybridization. The amplification of DNA (PCR) or a cDNA (RT-PCR) in cell or tissue sections has been developed at light and electron microscopic levels. A successful PCR experiment requires the careful optimization of several parameters depending on the tissue (or of cell types), and a compromise must be found between the fixation time, pretreatments and a good preservation of the morphology. Other crucial factors (primer design, concentration in MgCl2, annealing and elongation temperatures during the amplification steps) and their influence on the specificity and sensitivity of in situ PCR or RT-PCR are discussed. The necessity to run appropriate controls, especially to assess the lack of diffusion of the amplified products, is stressed. Current applications and future trends are also presented.</div></front></TEI><istex><corpusName>elsevier</corpusName><keywords><teeft><json:string>polymerase</json:string><json:string>hybridization</json:string><json:string>primer</json:string><json:string>mrna</json:string><json:string>transcriptase</json:string><json:string>komminoth</json:string><json:string>receptor</json:string><json:string>cdna</json:string><json:string>polymerase chain reaction</json:string><json:string>fixation</json:string><json:string>viral</json:string><json:string>amplification</json:string><json:string>tissue sections</json:string><json:string>digestion</json:string><json:string>morel</json:string><json:string>ultrathin</json:string><json:string>nuovo</json:string><json:string>fixative</json:string><json:string>dntps</json:string><json:string>growth hormone receptor mrna</json:string><json:string>denaturation</json:string><json:string>genome</json:string><json:string>embedding</json:string><json:string>bagasra</json:string><json:string>mertani</json:string><json:string>ronsin</json:string><json:string>nucleic</json:string><json:string>dnase</json:string><json:string>paraformaldehyde</json:string><json:string>rnase</json:string><json:string>mispriming</json:string><json:string>annealing</json:string><json:string>nucleic acids</json:string><json:string>haase</json:string><json:string>electron microscopy</json:string><json:string>ethanol</json:string><json:string>nucleotide</json:string><json:string>first step</json:string><json:string>boehringer mannheim</json:string><json:string>polymerase activity</json:string><json:string>gold particles</json:string><json:string>cycle number</json:string><json:string>positive cells</json:string><json:string>replication</json:string><json:string>negative strand</json:string><json:string>viral methods</json:string><json:string>nucleic acid</json:string><json:string>infectious agents</json:string><json:string>appropriate controls</json:string><json:string>dnase digestion</json:string><json:string>vibratome sections</json:string><json:string>target sequence</json:string><json:string>second step</json:string><json:string>cell membrane</json:string><json:string>proteolytic digestion</json:string><json:string>human press</json:string><json:string>hybridization step</json:string><json:string>alkaline phosphatase</json:string><json:string>boca york</json:string><json:string>phosphate buffer</json:string><json:string>room temperature</json:string><json:string>molecular biology</json:string><json:string>glass slides</json:string><json:string>protocol</json:string><json:string>localization</json:string><json:string>powerful technique</json:string><json:string>elongation step</json:string><json:string>endogenous biotin</json:string><json:string>amplification mixture</json:string><json:string>several approaches</json:string><json:string>humid chamber</json:string><json:string>direct reaction</json:string><json:string>unlabeled nucleotides</json:string><json:string>possible degradation</json:string><json:string>amplification reaction</json:string><json:string>major pitfall</json:string><json:string>unlabeled probe</json:string><json:string>pituitary gland section</json:string><json:string>high number</json:string><json:string>indirect reactions</json:string><json:string>digestion step</json:string><json:string>secondary structures</json:string><json:string>nucleic acid target</json:string><json:string>fixation time</json:string><json:string>light microscopy</json:string><json:string>amplification products</json:string><json:string>host cell</json:string><json:string>viral genome</json:string><json:string>positive strand</json:string><json:string>tissue section</json:string><json:string>final concentration</json:string><json:string>biological material</json:string><json:string>possible adsorption</json:string><json:string>avian myeloblastosis virus</json:string><json:string>total volume</json:string><json:string>extension time</json:string><json:string>hybridization temperature</json:string><json:string>mrna coding</json:string><json:string>endocrine cell populations</json:string><json:string>higher concentration</json:string><json:string>hydrophylic resin</json:string><json:string>human papillomavirus</json:string><json:string>signal density</json:string><json:string>visna virus</json:string><json:string>white resin</json:string><json:string>ultrathin sections</json:string><json:string>tissue preparation</json:string><json:string>nucleic acid sequences</json:string><json:string>diffusion artifacts</json:string><json:string>gene expression</json:string><json:string>molecular diagnosis</json:string><json:string>good preservation</json:string><json:string>careful optimization</json:string><json:string>gene rearrangements</json:string><json:string>cell suspensions</json:string><json:string>raven press</json:string><json:string>individual cells</json:string><json:string>detection system</json:string></teeft></keywords><author><json:item><name>Gérard Morel</name><affiliations><json:string>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</json:string></affiliations></json:item><json:item><name>Martina Berger</name><affiliations><json:string>Laboratoire de Virologie, Faculté de Médecine, Domaine Rockefeller, Lyon, France</json:string></affiliations></json:item><json:item><name>Brice Ronsin</name><affiliations><json:string>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</json:string></affiliations></json:item><json:item><name>Sophie Recher</name><affiliations><json:string>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</json:string></affiliations></json:item><json:item><name>Sylvie Ricard-Blum</name><affiliations><json:string>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</json:string></affiliations></json:item><json:item><name>Hichem C Mertani</name><affiliations><json:string>Institute of Molecular and Cell Biology, National University of Singapore, Singapore</json:string></affiliations></json:item><json:item><name>Peter E Lobie</name><affiliations><json:string>Institute of Molecular and Cell Biology, National University of Singapore, Singapore</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>in situ RT-PCR light microscopy</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>electron microscopy</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>growth hormone receptor mRNA</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>virus amplification</value></json:item></subject><arkIstex>ark:/67375/6H6-G281DK2K-F</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>Review article</json:string></originalGenre><abstract>Abstract: Since its discovery in 1986 by Mullis, the polymerase chain reaction (PCR) has been extensively developed by morphologists in order to overcome the main limitation of in situ hybridization, the lack of sensitivity. In situ PCR combines the extreme sensitivity of PCR with the cell-localizing ability of in situ hybridization. The amplification of DNA (PCR) or a cDNA (RT-PCR) in cell or tissue sections has been developed at light and electron microscopic levels. A successful PCR experiment requires the careful optimization of several parameters depending on the tissue (or of cell types), and a compromise must be found between the fixation time, pretreatments and a good preservation of the morphology. Other crucial factors (primer design, concentration in MgCl2, annealing and elongation temperatures during the amplification steps) and their influence on the specificity and sensitivity of in situ PCR or RT-PCR are discussed. The necessity to run appropriate controls, especially to assess the lack of diffusion of the amplified products, is stressed. Current applications and future trends are also presented.</abstract><qualityIndicators><score>9.04</score><pdfWordCount>9503</pdfWordCount><pdfCharCount>59944</pdfCharCount><pdfVersion>1.3</pdfVersion><pdfPageCount>18</pdfPageCount><pdfPageSize>614 x 864 pts</pdfPageSize><refBibsNative>true</refBibsNative><abstractWordCount>170</abstractWordCount><abstractCharCount>1128</abstractCharCount><keywordCount>4</keywordCount></qualityIndicators><title>In situ reverse transcription-polymerase chain reaction. Applications for light and electron microscopy</title><pmid><json:string>9691431</json:string></pmid><pii><json:string>S0248-4900(98)80335-3</json:string></pii><genre><json:string>review-article</json:string></genre><serie><title>Proc Natl Acad Sci USA</title><language><json:string>unknown</json:string></language><volume>90</volume><pages><first>359</first><last>362</last></pages></serie><host><title>Biology of the Cell</title><language><json:string>unknown</json:string></language><publicationDate>1998</publicationDate><issn><json:string>0248-4900</json:string></issn><pii><json:string>S0248-4900(00)X0018-4</json:string></pii><volume>90</volume><issue>2</issue><pages><first>137</first><last>154</last></pages><genre><json:string>journal</json:string></genre></host><namedEntities><unitex><date><json:string>1986</json:string><json:string>between 22s and 23s</json:string><json:string>1969</json:string><json:string>1000</json:string><json:string>1998</json:string><json:string>1840</json:string><json:string>1990</json:string></date><geogName></geogName><orgName><json:string>Boehringer Mannheim, Germany</json:string><json:string>Human Press Inc</json:string><json:string>Cytochem</json:string><json:string>Histochem</json:string><json:string>National University of Singapore, Singapore Since</json:string><json:string>Institute of Molecular and Cell Biology</json:string><json:string>Biochem</json:string><json:string>Perkin Elmer</json:string><json:string>Boehringer Mannheim</json:string></orgName><orgName_funder></orgName_funder><orgName_provider></orgName_provider><persName><json:string>Jacob Henle</json:string><json:string>La Doua</json:string><json:string>NY Acad</json:string><json:string>Raven Press</json:string><json:string>Mel Med</json:string><json:string>A. Method</json:string><json:string>Sylvie Ricard</json:string><json:string>Spring Harbor</json:string><json:string>Viral Methods</json:string><json:string>Raven Publishers</json:string><json:string>I. Principle</json:string><json:string>Lab</json:string><json:string>Bernard C. Mougin</json:string><json:string>Mel Pathol</json:string></persName><placeName><json:string>Wakefield</json:string><json:string>Linz</json:string><json:string>Austria</json:string><json:string>Prato</json:string><json:string>Vienna</json:string><json:string>New Engl</json:string><json:string>York</json:string><json:string>Totowa</json:string><json:string>France</json:string><json:string>NJ</json:string><json:string>Lyon</json:string></placeName><ref_url></ref_url><ref_bibl><json:string>Nuovo et al, 1991a, b</json:string><json:string>Chomczynski and Sac&i, 1993</json:string><json:string>Maher et al, 1995</json:string><json:string>Ogretmen and Safa, 1995</json:string><json:string>Bernard et d, 1994</json:string><json:string>Higuchi and Kwok, 1989</json:string><json:string>Embretson el al, 1993</json:string><json:string>Chiu et al, 1993</json:string><json:string>Murray et al, 1994</json:string><json:string>Komminoth and Long, 1995</json:string><json:string>Prato-Murphy ef al, 1995</json:string><json:string>Morel, 1991, 1993</json:string><json:string>Chattopadhyay et al, 1993</json:string><json:string>Patterson et al, 1993</json:string><json:string>Linz et al, 1989</json:string><json:string>Grandclement et al (1997)</json:string><json:string>Mertani et al, 1994</json:string><json:string>Pate1 et al, 1994</json:string><json:string>Ray et al, 1991</json:string><json:string>Nuovo et al, 1993</json:string><json:string>Hayes and Buck, 1993</json:string><json:string>Nuovo and Richart, 1989</json:string><json:string>Jackson et al, 1990</json:string><json:string>Ronsin et al, 1995</json:string><json:string>Embleton et al, 1992</json:string><json:string>Mertani and Morel, 1995</json:string><json:string>Santa and Schneider, 1991</json:string><json:string>Ronsin and Morel, 1997</json:string><json:string>Greer et al, 1994</json:string><json:string>1993, 1995</json:string><json:string>Haase et al, 1990</json:string><json:string>Weirich et al, 1995</json:string><json:string>Long et al, 1993</json:string><json:string>Troxler et al, 1990</json:string><json:string>Mertani ef al, 1995</json:string><json:string>Zoccoli, 1994</json:string><json:string>Pardue and Gall (1969)</json:string><json:string>Haase et al</json:string><json:string>Komminoth and Long, 1993</json:string><json:string>Runsin and Morel, 1997</json:string><json:string>Mertani and Morel, 1994</json:string><json:string>Kelly et al, 1991</json:string><json:string>Isaacson et al, 1995</json:string><json:string>NUOVO, 1994</json:string><json:string>Roizman and Palese, 1996</json:string><json:string>Komminoth and Long, 1993, 1995</json:string><json:string>Tokuyasu, 1973</json:string><json:string>Banejee et al, 1995</json:string><json:string>cf fig 70 and Long, 1993</json:string><json:string>Bagasra et al, 1993a</json:string><json:string>Grandclement ef al, 1997</json:string><json:string>Tecott et al (1988)</json:string><json:string>Komminoth et al, 1994</json:string><json:string>Pate1 ef al, 1994</json:string><json:string>Nuovo, 1992c</json:string><json:string>Bagasra et al, 1992</json:string><json:string>Nuovo, 1992a, b</json:string><json:string>Vauti and Siess, 1993</json:string><json:string>Bagasra et al, 1997</json:string><json:string>kguellec et al, 1992</json:string><json:string>Ray et ~2, 1996</json:string><json:string>for review see Nuovo, 1997</json:string><json:string>Hodges and Smith, 1995</json:string><json:string>Berger et al, 1997</json:string><json:string>Morel et al</json:string><json:string>Haase et al (1990)</json:string><json:string>Embretson et al, 1993</json:string><json:string>Murray et aZ, 1994</json:string><json:string>Stackus et al, 1991</json:string><json:string>Haase ef al, 1990</json:string><json:string>Berger et aI, 1997</json:string><json:string>Nuovo et a2,1993</json:string><json:string>Chen and Fuggle, 1993</json:string><json:string>Billstrom et al, 1993</json:string><json:string>Teo and Shaunak, 1995</json:string><json:string>Nuovo, 1994,1997</json:string><json:string>Cheng and Nuovo, 1994</json:string><json:string>Morei et al</json:string><json:string>Groppe and Morse, 1993</json:string><json:string>Nuovo, 1997</json:string><json:string>Span et uZ, 1991</json:string><json:string>Ronsin et a2, 1995</json:string><json:string>Komminoth et al, 1992a, b, 1994</json:string></ref_bibl><bibl></bibl></unitex></namedEntities><ark><json:string>ark:/67375/6H6-G281DK2K-F</json:string></ark><categories><wos><json:string>1 - 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Applications for light and electron microscopy</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher scheme="https://scientific-publisher.data.istex.fr">ELSEVIER</publisher><availability><licence><p>elsevier</p></licence></availability><p scheme="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M"></p><date>1998</date></publicationStmt><notesStmt><note type="review-article" scheme="https://content-type.data.istex.fr/ark:/67375/XTP-L5L7X3NF-P">review-article</note><note type="journal" scheme="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</note><note type="content">Section title: Review</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a">In situ reverse transcription-polymerase chain reaction. Applications for light and electron microscopy</title><author xml:id="author-0000"><persName><forename type="first">Gérard</forename><surname>Morel</surname></persName><affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</affiliation></author><author xml:id="author-0001"><persName><forename type="first">Martina</forename><surname>Berger</surname></persName><affiliation>Laboratoire de Virologie, Faculté de Médecine, Domaine Rockefeller, Lyon, France</affiliation></author><author xml:id="author-0002"><persName><forename type="first">Brice</forename><surname>Ronsin</surname></persName><affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</affiliation></author><author xml:id="author-0003"><persName><forename type="first">Sophie</forename><surname>Recher</surname></persName><affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</affiliation></author><author xml:id="author-0004"><persName><forename type="first">Sylvie</forename><surname>Ricard-Blum</surname></persName><affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</affiliation></author><author xml:id="author-0005"><persName><forename type="first">Hichem C</forename><surname>Mertani</surname></persName><affiliation>Institute of Molecular and Cell Biology, National University of Singapore, Singapore</affiliation></author><author xml:id="author-0006"><persName><forename type="first">Peter E</forename><surname>Lobie</surname></persName><affiliation>Institute of Molecular and Cell Biology, National University of Singapore, Singapore</affiliation></author><idno type="istex">7DC11D66EC5B5B3B9875983BDAFD1A98F370D651</idno><idno type="ark">ark:/67375/6H6-G281DK2K-F</idno><idno type="DOI">10.1016/S0248-4900(98)80335-3</idno><idno type="PII">S0248-4900(98)80335-3</idno></analytic><monogr><title level="j">Biology of the Cell</title><title level="j" type="abbrev">BICELL</title><idno type="pISSN">0248-4900</idno><idno type="PII">S0248-4900(00)X0018-4</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1998"></date><biblScope unit="volume">90</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="137">137</biblScope><biblScope unit="page" to="154">154</biblScope></imprint></monogr></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1998</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Abstract: Since its discovery in 1986 by Mullis, the polymerase chain reaction (PCR) has been extensively developed by morphologists in order to overcome the main limitation of in situ hybridization, the lack of sensitivity. In situ PCR combines the extreme sensitivity of PCR with the cell-localizing ability of in situ hybridization. The amplification of DNA (PCR) or a cDNA (RT-PCR) in cell or tissue sections has been developed at light and electron microscopic levels. A successful PCR experiment requires the careful optimization of several parameters depending on the tissue (or of cell types), and a compromise must be found between the fixation time, pretreatments and a good preservation of the morphology. Other crucial factors (primer design, concentration in MgCl2, annealing and elongation temperatures during the amplification steps) and their influence on the specificity and sensitivity of in situ PCR or RT-PCR are discussed. The necessity to run appropriate controls, especially to assess the lack of diffusion of the amplified products, is stressed. Current applications and future trends are also presented.</p></abstract><textClass><keywords scheme="keyword"><list><head>Keywords</head><item><term>in situ RT-PCR light microscopy</term></item><item><term>electron microscopy</term></item><item><term>growth hormone receptor mRNA</term></item><item><term>virus amplification</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1998">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-G281DK2K-F/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: tail"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="rev"><item-info><jid>BICELL</jid><aid>98803353</aid><ce:pii>S0248-4900(98)80335-3</ce:pii><ce:doi>10.1016/S0248-4900(98)80335-3</ce:doi><ce:copyright type="unknown" year="1998"></ce:copyright></item-info><head><ce:dochead><ce:textfn>Review</ce:textfn></ce:dochead><ce:title><ce:italic>In situ</ce:italic> reverse transcription-polymerase chain reaction. Applications for light and electron microscopy</ce:title><ce:author-group><ce:author><ce:given-name>Gérard</ce:given-name><ce:surname>Morel</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Martina</ce:given-name><ce:surname>Berger</ce:surname><ce:cross-ref refid="AFF2"><ce:sup>b</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Brice</ce:given-name><ce:surname>Ronsin</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Sophie</ce:given-name><ce:surname>Recher</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Sylvie</ce:given-name><ce:surname>Ricard-Blum</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Hichem C</ce:given-name><ce:surname>Mertani</ce:surname><ce:cross-ref refid="AFF3"><ce:sup>c</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Peter E</ce:given-name><ce:surname>Lobie</ce:surname><ce:cross-ref refid="AFF3"><ce:sup>c</ce:sup></ce:cross-ref></ce:author><ce:affiliation id="AFF1"><ce:label>a</ce:label><ce:textfn>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3<ce:sup>e</ce:sup> étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</ce:textfn></ce:affiliation><ce:affiliation id="AFF2"><ce:label>b</ce:label><ce:textfn>Laboratoire de Virologie, Faculté de Médecine, Domaine Rockefeller, Lyon, France</ce:textfn></ce:affiliation><ce:affiliation id="AFF3"><ce:label>c</ce:label><ce:textfn>Institute of Molecular and Cell Biology, National University of Singapore, Singapore</ce:textfn></ce:affiliation></ce:author-group><ce:date-received day="22" month="1" year="1998"></ce:date-received><ce:date-accepted day="3" month="3" year="1998"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>Since its discovery in 1986 by Mullis, the polymerase chain reaction (PCR) has been extensively developed by morphologists in order to overcome the main limitation of <ce:italic>in situ</ce:italic> hybridization, the lack of sensitivity. <ce:italic>In situ</ce:italic> PCR combines the extreme sensitivity of PCR with the cell-localizing ability of <ce:italic>in situ</ce:italic> hybridization. The amplification of DNA (PCR) or a cDNA (RT-PCR) in cell or tissue sections has been developed at light and electron microscopic levels. A successful PCR experiment requires the careful optimization of several parameters depending on the tissue (or of cell types), and a compromise must be found between the fixation time, pretreatments and a good preservation of the morphology. Other crucial factors (primer design, concentration in MgCl<ce:inf>2</ce:inf>, annealing and elongation temperatures during the amplification steps) and their influence on the specificity and sensitivity of <ce:italic>in situ</ce:italic> PCR or RT-PCR are discussed. The necessity to run appropriate controls, especially to assess the lack of diffusion of the amplified products, is stressed. Current applications and future trends are also presented.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text><ce:italic>in situ</ce:italic> RT-PCR light microscopy</ce:text></ce:keyword><ce:keyword><ce:text>electron microscopy</ce:text></ce:keyword><ce:keyword><ce:text>growth hormone receptor mRNA</ce:text></ce:keyword><ce:keyword><ce:text>virus amplification</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>In situ reverse transcription-polymerase chain reaction. Applications for light and electron microscopy</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>reverse transcription-polymerase chain reaction. Applications for light and electron microscopy</title></titleInfo><name type="personal"><namePart type="given">Gérard</namePart><namePart type="family">Morel</namePart><affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Martina</namePart><namePart type="family">Berger</namePart><affiliation>Laboratoire de Virologie, Faculté de Médecine, Domaine Rockefeller, Lyon, France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Brice</namePart><namePart type="family">Ronsin</namePart><affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Sophie</namePart><namePart type="family">Recher</namePart><affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Sylvie</namePart><namePart type="family">Ricard-Blum</namePart><affiliation>CNRS-UMR 5578, Université Claude-Bernard-Lyon 1, bât 404, 3e étage, Campus de La Doua, 43, blvd du 11-Novembre-1918, 69622 Villeurbanne Cedex France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Hichem C</namePart><namePart type="family">Mertani</namePart><affiliation>Institute of Molecular and Cell Biology, National University of Singapore, Singapore</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Peter E</namePart><namePart type="family">Lobie</namePart><affiliation>Institute of Molecular and Cell Biology, National University of Singapore, Singapore</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="review-article" displayLabel="Review article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-L5L7X3NF-P">review-article</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1998</dateIssued><copyrightDate encoding="w3cdtf">1998</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Abstract: Since its discovery in 1986 by Mullis, the polymerase chain reaction (PCR) has been extensively developed by morphologists in order to overcome the main limitation of in situ hybridization, the lack of sensitivity. In situ PCR combines the extreme sensitivity of PCR with the cell-localizing ability of in situ hybridization. The amplification of DNA (PCR) or a cDNA (RT-PCR) in cell or tissue sections has been developed at light and electron microscopic levels. A successful PCR experiment requires the careful optimization of several parameters depending on the tissue (or of cell types), and a compromise must be found between the fixation time, pretreatments and a good preservation of the morphology. Other crucial factors (primer design, concentration in MgCl2, annealing and elongation temperatures during the amplification steps) and their influence on the specificity and sensitivity of in situ PCR or RT-PCR are discussed. The necessity to run appropriate controls, especially to assess the lack of diffusion of the amplified products, is stressed. Current applications and future trends are also presented.</abstract><note type="content">Section title: Review</note><subject><genre>Keywords</genre><topic>in situ RT-PCR light microscopy</topic><topic>electron microscopy</topic><topic>growth hormone receptor mRNA</topic><topic>virus amplification</topic></subject><relatedItem type="host"><titleInfo><title>Biology of the Cell</title></titleInfo><titleInfo type="abbreviated"><title>BICELL</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1998</dateIssued></originInfo><identifier type="ISSN">0248-4900</identifier><identifier type="PII">S0248-4900(00)X0018-4</identifier><part><date>1998</date><detail type="volume"><number>90</number><caption>vol.</caption></detail><detail type="issue"><number>2</number><caption>no.</caption></detail><extent unit="issue-pages"><start>137</start><end>205</end></extent><extent unit="pages"><start>137</start><end>154</end></extent></part></relatedItem><identifier type="istex">7DC11D66EC5B5B3B9875983BDAFD1A98F370D651</identifier><identifier type="ark">ark:/67375/6H6-G281DK2K-F</identifier><identifier type="DOI">10.1016/S0248-4900(98)80335-3</identifier><identifier type="PII">S0248-4900(98)80335-3</identifier><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</recordContentSource></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-G281DK2K-F/record.json</uri></json:item></metadata></istex></record>Pour manipuler ce document sous Unix (Dilib)
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