Single-Cell Quantification of mRNA Expression in The Human Brain
RNA analysis at the cellular resolution in the human brain is challenging. Here, we describe an optimised approach for detecting single RNA transcripts in a cell-type specific manner in frozen human brain tissue using multiplexed fluorescent RNAscope probes. We developed a new robust analytical appr...
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| Veröffentlicht in: | Scientific reports 2019-08, Vol.9 (1), p.12353-9, Article 12353 |
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| creator | Jolly, Sarah Lang, Verena Koelzer, Viktor Hendrik Sala Frigerio, Carlo Magno, Lorenza Salinas, Patricia C. Whiting, Paul Palomer, Ernest |
| description | RNA analysis at the cellular resolution in the human brain is challenging. Here, we describe an optimised approach for detecting single RNA transcripts in a cell-type specific manner in frozen human brain tissue using multiplexed fluorescent RNAscope probes. We developed a new robust analytical approach for RNAscope quantification. Our method shows that low RNA integrity does not significantly affect RNAscope signal, recapitulates bulk RNA analysis and provides spatial context to transcriptomic analysis of human post-mortem brain at single-cell resolution. In summary, our optimised method allows the usage of frozen human samples from brain banks to perform quantitative RNAscope analysis. |
| doi_str_mv | 10.1038/s41598-019-48787-w |
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Here, we describe an optimised approach for detecting single RNA transcripts in a cell-type specific manner in frozen human brain tissue using multiplexed fluorescent RNAscope probes. We developed a new robust analytical approach for RNAscope quantification. Our method shows that low RNA integrity does not significantly affect RNAscope signal, recapitulates bulk RNA analysis and provides spatial context to transcriptomic analysis of human post-mortem brain at single-cell resolution. In summary, our optimised method allows the usage of frozen human samples from brain banks to perform quantitative RNAscope analysis.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/s41598-019-48787-w</identifier><identifier>PMID: 31451701</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>14/32 ; 38/32 ; 38/77 ; 631/337/572 ; 631/378/340 ; Alzheimer Disease - genetics ; Brain ; Brain - metabolism ; DNA probes ; Fluorescent indicators ; Freezing ; Gene expression ; Gene Expression Regulation ; HeLa Cells ; Hippocampus - metabolism ; Hippocampus - pathology ; Humanities and Social Sciences ; Humans ; Membrane Glycoproteins - genetics ; Membrane Glycoproteins - metabolism ; Microglia - metabolism ; multidisciplinary ; Receptors, Immunologic - genetics ; Receptors, Immunologic - metabolism ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; Science ; Science (multidisciplinary) ; Single-Cell Analysis ; Spatial analysis ; Up-Regulation</subject><ispartof>Scientific reports, 2019-08, Vol.9 (1), p.12353-9, Article 12353</ispartof><rights>The Author(s) 2019</rights><rights>2019. 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Here, we describe an optimised approach for detecting single RNA transcripts in a cell-type specific manner in frozen human brain tissue using multiplexed fluorescent RNAscope probes. We developed a new robust analytical approach for RNAscope quantification. Our method shows that low RNA integrity does not significantly affect RNAscope signal, recapitulates bulk RNA analysis and provides spatial context to transcriptomic analysis of human post-mortem brain at single-cell resolution. 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jolly, Sarah</au><au>Lang, Verena</au><au>Koelzer, Viktor Hendrik</au><au>Sala Frigerio, Carlo</au><au>Magno, Lorenza</au><au>Salinas, Patricia C.</au><au>Whiting, Paul</au><au>Palomer, Ernest</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Single-Cell Quantification of mRNA Expression in The Human Brain</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2019-08-26</date><risdate>2019</risdate><volume>9</volume><issue>1</issue><spage>12353</spage><epage>9</epage><pages>12353-9</pages><artnum>12353</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>RNA analysis at the cellular resolution in the human brain is challenging. 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| subjects | 14/32 38/32 38/77 631/337/572 631/378/340 Alzheimer Disease - genetics Brain Brain - metabolism DNA probes Fluorescent indicators Freezing Gene expression Gene Expression Regulation HeLa Cells Hippocampus - metabolism Hippocampus - pathology Humanities and Social Sciences Humans Membrane Glycoproteins - genetics Membrane Glycoproteins - metabolism Microglia - metabolism multidisciplinary Receptors, Immunologic - genetics Receptors, Immunologic - metabolism RNA, Messenger - genetics RNA, Messenger - metabolism Science Science (multidisciplinary) Single-Cell Analysis Spatial analysis Up-Regulation |
| title | Single-Cell Quantification of mRNA Expression in The Human Brain |
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