Turbo FISH: a method for rapid single molecule RNA FISH.
Advances in RNA fluorescence in situ hybridization (RNA FISH) have allowed practitioners to detect individual RNA molecules in single cells via fluorescence microscopy, enabling highly accurate and sensitive quantification of gene expression. However, current methods typically employ hybridization t...
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ftdoajarticles:oai:doaj.org/article:7746527def4e4439825711fee6868063 2023-05-15T16:42:07+02:00 Turbo FISH: a method for rapid single molecule RNA FISH. Sydney M Shaffer Min-Tzu Wu Marshall J Levesque Arjun Raj 2013-01-01T00:00:00Z https://doi.org/10.1371/journal.pone.0075120 https://doaj.org/article/7746527def4e4439825711fee6868063 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3774626?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0075120 https://doaj.org/article/7746527def4e4439825711fee6868063 PLoS ONE, Vol 8, Iss 9, p e75120 (2013) Medicine R Science Q article 2013 ftdoajarticles https://doi.org/10.1371/journal.pone.0075120 2022-12-31T12:25:24Z Advances in RNA fluorescence in situ hybridization (RNA FISH) have allowed practitioners to detect individual RNA molecules in single cells via fluorescence microscopy, enabling highly accurate and sensitive quantification of gene expression. However, current methods typically employ hybridization times on the order of 2-16 hours, limiting its potential in applications like rapid diagnostics. We present here a set of conditions for RNA FISH (dubbed Turbo RNA FISH) that allow us to make accurate measurements with no more than 5 minutes of hybridization time and 3 minutes of washing, and show that hybridization times can go as low as 30 seconds while still producing quantifiable images. We further show that rapid hybridization is compatible with our recently developed iceFISH and SNP FISH variants of RNA FISH that enable chromosome and single base discrimination, respectively. Our method is simple and cost effective, and has the potential to dramatically increase the throughput and realm of applicability of RNA FISH. Article in Journal/Newspaper Icefish Directory of Open Access Journals: DOAJ Articles PLoS ONE 8 9 e75120 |
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Directory of Open Access Journals: DOAJ Articles |
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Medicine R Science Q |
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Medicine R Science Q Sydney M Shaffer Min-Tzu Wu Marshall J Levesque Arjun Raj Turbo FISH: a method for rapid single molecule RNA FISH. |
topic_facet |
Medicine R Science Q |
description |
Advances in RNA fluorescence in situ hybridization (RNA FISH) have allowed practitioners to detect individual RNA molecules in single cells via fluorescence microscopy, enabling highly accurate and sensitive quantification of gene expression. However, current methods typically employ hybridization times on the order of 2-16 hours, limiting its potential in applications like rapid diagnostics. We present here a set of conditions for RNA FISH (dubbed Turbo RNA FISH) that allow us to make accurate measurements with no more than 5 minutes of hybridization time and 3 minutes of washing, and show that hybridization times can go as low as 30 seconds while still producing quantifiable images. We further show that rapid hybridization is compatible with our recently developed iceFISH and SNP FISH variants of RNA FISH that enable chromosome and single base discrimination, respectively. Our method is simple and cost effective, and has the potential to dramatically increase the throughput and realm of applicability of RNA FISH. |
format |
Article in Journal/Newspaper |
author |
Sydney M Shaffer Min-Tzu Wu Marshall J Levesque Arjun Raj |
author_facet |
Sydney M Shaffer Min-Tzu Wu Marshall J Levesque Arjun Raj |
author_sort |
Sydney M Shaffer |
title |
Turbo FISH: a method for rapid single molecule RNA FISH. |
title_short |
Turbo FISH: a method for rapid single molecule RNA FISH. |
title_full |
Turbo FISH: a method for rapid single molecule RNA FISH. |
title_fullStr |
Turbo FISH: a method for rapid single molecule RNA FISH. |
title_full_unstemmed |
Turbo FISH: a method for rapid single molecule RNA FISH. |
title_sort |
turbo fish: a method for rapid single molecule rna fish. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2013 |
url |
https://doi.org/10.1371/journal.pone.0075120 https://doaj.org/article/7746527def4e4439825711fee6868063 |
genre |
Icefish |
genre_facet |
Icefish |
op_source |
PLoS ONE, Vol 8, Iss 9, p e75120 (2013) |
op_relation |
http://europepmc.org/articles/PMC3774626?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0075120 https://doaj.org/article/7746527def4e4439825711fee6868063 |
op_doi |
https://doi.org/10.1371/journal.pone.0075120 |
container_title |
PLoS ONE |
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8 |
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9 |
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e75120 |
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1766032562576162816 |