Gene－expression analysis of single cells－nested polymerase chain reaction after laser microdissection
AIM: The structural and functional characteristics of cells are dependent on the specific gene expression profile. The ability to study and compare gene expression at the cellular level will therefore provide valuable insights into cell physiology and pathophysiology.METHODS: Individual cells were i...
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| Veröffentlicht in: | World journal of gastroenterology : WJG 2003-06, Vol.9 (6), p.1337-1341 |
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| container_title | World journal of gastroenterology : WJG |
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| creator | Shi, Xin Kleeff, Jörg Zhu, Zhao-Wen Schmied, Bruno Tang, Wen-Hao Zimmermann, Arthur Buchler, Markus W Friess, Helmut |
| description | AIM: The structural and functional characteristics of cells are dependent on the specific gene expression profile. The ability to study and compare gene expression at the cellular level will therefore provide valuable insights into cell physiology and pathophysiology.METHODS: Individual cells were isolated from frozen colon tissue sections using laser microdissection. DNA as well as RNA were extracted, and total RNA was reversely transcribed to complementary DNA (cDNA). Both DNA and cDNA were analyzed by nested polymerase chain reaction (PCR). The quality of isolated DNA and RNA was satisfactory.RESULTS: Single cells were successfully microdissected using an ultraviolet laser micromanipulator. Nested PCR amplification products of DNA and cDNA of single cells could clearly be visualized by agarose gel electrophoresis.CONCLUSION: The combined use of laser microdissection and nested-PCR provides an opportunity to analyze gene expression in single cells. This method allows the analysis and identification of specific genes which are in volved inphysiological and pathophysiological processes in a complex of variable cell phenotypes. |
| doi_str_mv | 10.3748/wjg.v9.i6.1337 |
| format | Article |
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The ability to study and compare gene expression at the cellular level will therefore provide valuable insights into cell physiology and pathophysiology.METHODS: Individual cells were isolated from frozen colon tissue sections using laser microdissection. DNA as well as RNA were extracted, and total RNA was reversely transcribed to complementary DNA (cDNA). Both DNA and cDNA were analyzed by nested polymerase chain reaction (PCR). The quality of isolated DNA and RNA was satisfactory.RESULTS: Single cells were successfully microdissected using an ultraviolet laser micromanipulator. Nested PCR amplification products of DNA and cDNA of single cells could clearly be visualized by agarose gel electrophoresis.CONCLUSION: The combined use of laser microdissection and nested-PCR provides an opportunity to analyze gene expression in single cells. This method allows the analysis and identification of specific genes which are in volved inphysiological and pathophysiological processes in a complex of variable cell phenotypes.</description><identifier>ISSN: 1007-9327</identifier><identifier>EISSN: 2219-2840</identifier><identifier>DOI: 10.3748/wjg.v9.i6.1337</identifier><identifier>PMID: 12800252</identifier><language>eng</language><publisher>United States: Baishideng Publishing Group Inc</publisher><subject>Basic Research ; Colon - cytology ; Colon - physiology ; Colon - surgery ; Dissection ; Gene Expression ; Humans ; Laser Therapy ; Polymerase Chain Reaction ; 基因表达 ; 病理生理学 ; 细胞生理学 ; 聚合酶链反应</subject><ispartof>World journal of gastroenterology : WJG, 2003-06, Vol.9 (6), p.1337-1341</ispartof><rights>The Author(s) 2003. Published by Baishideng Publishing Group Inc. All rights reserved. 2003</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c478t-f966485c6a0cabc91233aa2aa8128643d9d7faac17db490c52b70aafe6acdec63</citedby><cites>FETCH-LOGICAL-c478t-f966485c6a0cabc91233aa2aa8128643d9d7faac17db490c52b70aafe6acdec63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://image.cqvip.com/vip1000/qk/84123X/84123X.jpg</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4611812/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4611812/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12800252$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Shi, Xin</creatorcontrib><creatorcontrib>Kleeff, Jörg</creatorcontrib><creatorcontrib>Zhu, Zhao-Wen</creatorcontrib><creatorcontrib>Schmied, Bruno</creatorcontrib><creatorcontrib>Tang, Wen-Hao</creatorcontrib><creatorcontrib>Zimmermann, Arthur</creatorcontrib><creatorcontrib>Buchler, Markus W</creatorcontrib><creatorcontrib>Friess, Helmut</creatorcontrib><title>Gene－expression analysis of single cells－nested polymerase chain reaction after laser microdissection</title><title>World journal of gastroenterology : WJG</title><addtitle>World Journal of Gastroenterology</addtitle><description>AIM: The structural and functional characteristics of cells are dependent on the specific gene expression profile. The ability to study and compare gene expression at the cellular level will therefore provide valuable insights into cell physiology and pathophysiology.METHODS: Individual cells were isolated from frozen colon tissue sections using laser microdissection. DNA as well as RNA were extracted, and total RNA was reversely transcribed to complementary DNA (cDNA). Both DNA and cDNA were analyzed by nested polymerase chain reaction (PCR). The quality of isolated DNA and RNA was satisfactory.RESULTS: Single cells were successfully microdissected using an ultraviolet laser micromanipulator. Nested PCR amplification products of DNA and cDNA of single cells could clearly be visualized by agarose gel electrophoresis.CONCLUSION: The combined use of laser microdissection and nested-PCR provides an opportunity to analyze gene expression in single cells. This method allows the analysis and identification of specific genes which are in volved inphysiological and pathophysiological processes in a complex of variable cell phenotypes.</description><subject>Basic Research</subject><subject>Colon - cytology</subject><subject>Colon - physiology</subject><subject>Colon - surgery</subject><subject>Dissection</subject><subject>Gene Expression</subject><subject>Humans</subject><subject>Laser Therapy</subject><subject>Polymerase Chain Reaction</subject><subject>基因表达</subject><subject>病理生理学</subject><subject>细胞生理学</subject><subject>聚合酶链反应</subject><issn>1007-9327</issn><issn>2219-2840</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVUU1v1DAUtKpWdFm49liFC7cEfyR2ckFCFRSkSlzK2XpxXnbdOvbWzi7sj-Af8Z_4C3jbFdCTZc28efNmCLlgtBKqbt99v1tVu66ysmJCqBOy4Jx1JW9rekoWjFJVdoKrc_IypTtKuRANf0HOGW_zp-ELYq_R4-9fP_HHJmJKNvgCPLh9sqkIY5GsXzksDDqXMstjmnEoNsHtJ4yQMrIG64uIYObH2XHGWLiMxGKyJobBpoSP2CtyNoJL-Pr4Lsm3Tx9vrz6XN1-vv1x9uClNrdq5HDsp67YxEqiB3nQsewbgAG02LWsxdIMaAQxTQ1931DS8VxRgRAlmQCPFkrx_0t1s-wkHg36O4PQm2gniXgew-jni7Vqvwk7XkrG8JAu8PQrE8LDNF-vJpkMC4DFsk1ZCSNbkuJekeiLmO1OKOP5dwqg-tKNzO3rXaSv1oZ08cPm_tX_0Yx2Z8OaouA5-9ZDD1z2Y-9E61C0TSkkq_gAdoJ7O</recordid><startdate>20030601</startdate><enddate>20030601</enddate><creator>Shi, Xin</creator><creator>Kleeff, Jörg</creator><creator>Zhu, Zhao-Wen</creator><creator>Schmied, Bruno</creator><creator>Tang, Wen-Hao</creator><creator>Zimmermann, Arthur</creator><creator>Buchler, Markus W</creator><creator>Friess, Helmut</creator><general>Baishideng Publishing Group Inc</general><scope>2RA</scope><scope>92L</scope><scope>CQIGP</scope><scope>W91</scope><scope>~WA</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20030601</creationdate><title>Gene－expression analysis of single cells－nested polymerase chain reaction after laser microdissection</title><author>Shi, Xin ; Kleeff, Jörg ; Zhu, Zhao-Wen ; Schmied, Bruno ; Tang, Wen-Hao ; Zimmermann, Arthur ; Buchler, Markus W ; Friess, Helmut</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c478t-f966485c6a0cabc91233aa2aa8128643d9d7faac17db490c52b70aafe6acdec63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Basic Research</topic><topic>Colon - cytology</topic><topic>Colon - physiology</topic><topic>Colon - surgery</topic><topic>Dissection</topic><topic>Gene Expression</topic><topic>Humans</topic><topic>Laser Therapy</topic><topic>Polymerase Chain Reaction</topic><topic>基因表达</topic><topic>病理生理学</topic><topic>细胞生理学</topic><topic>聚合酶链反应</topic><toplevel>online_resources</toplevel><creatorcontrib>Shi, Xin</creatorcontrib><creatorcontrib>Kleeff, Jörg</creatorcontrib><creatorcontrib>Zhu, Zhao-Wen</creatorcontrib><creatorcontrib>Schmied, Bruno</creatorcontrib><creatorcontrib>Tang, Wen-Hao</creatorcontrib><creatorcontrib>Zimmermann, Arthur</creatorcontrib><creatorcontrib>Buchler, Markus W</creatorcontrib><creatorcontrib>Friess, Helmut</creatorcontrib><collection>中文科技期刊数据库</collection><collection>中文科技期刊数据库-CALIS站点</collection><collection>中文科技期刊数据库-7.0平台</collection><collection>中文科技期刊数据库-医药卫生</collection><collection>中文科技期刊数据库- 镜像站点</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>World journal of gastroenterology : WJG</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Shi, Xin</au><au>Kleeff, Jörg</au><au>Zhu, Zhao-Wen</au><au>Schmied, Bruno</au><au>Tang, Wen-Hao</au><au>Zimmermann, Arthur</au><au>Buchler, Markus W</au><au>Friess, Helmut</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Gene－expression analysis of single cells－nested polymerase chain reaction after laser microdissection</atitle><jtitle>World journal of gastroenterology : WJG</jtitle><addtitle>World Journal of Gastroenterology</addtitle><date>2003-06-01</date><risdate>2003</risdate><volume>9</volume><issue>6</issue><spage>1337</spage><epage>1341</epage><pages>1337-1341</pages><issn>1007-9327</issn><eissn>2219-2840</eissn><abstract>AIM: The structural and functional characteristics of cells are dependent on the specific gene expression profile. The ability to study and compare gene expression at the cellular level will therefore provide valuable insights into cell physiology and pathophysiology.METHODS: Individual cells were isolated from frozen colon tissue sections using laser microdissection. DNA as well as RNA were extracted, and total RNA was reversely transcribed to complementary DNA (cDNA). Both DNA and cDNA were analyzed by nested polymerase chain reaction (PCR). The quality of isolated DNA and RNA was satisfactory.RESULTS: Single cells were successfully microdissected using an ultraviolet laser micromanipulator. Nested PCR amplification products of DNA and cDNA of single cells could clearly be visualized by agarose gel electrophoresis.CONCLUSION: The combined use of laser microdissection and nested-PCR provides an opportunity to analyze gene expression in single cells. 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| subjects | Basic Research Colon - cytology Colon - physiology Colon - surgery Dissection Gene Expression Humans Laser Therapy Polymerase Chain Reaction 基因表达 病理生理学 细胞生理学 聚合酶链反应 |
| title | Gene－expression analysis of single cells－nested polymerase chain reaction after laser microdissection |
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