Title |
Turbo FISH: A Method for Rapid Single Molecule RNA FISH
|
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Published in |
PLOS ONE, September 2013
|
DOI | 10.1371/journal.pone.0075120 |
Pubmed ID | |
Authors |
Sydney M. Shaffer, Min-Tzu Wu, Marshall J. Levesque, Arjun Raj |
Abstract |
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. |
X Demographics
Geographical breakdown
Country | Count | As % |
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United States | 1 | 25% |
France | 1 | 25% |
Unknown | 2 | 50% |
Demographic breakdown
Type | Count | As % |
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Scientists | 3 | 75% |
Members of the public | 1 | 25% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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United States | 8 | 3% |
United Kingdom | 3 | 1% |
Netherlands | 1 | <1% |
France | 1 | <1% |
Brazil | 1 | <1% |
Sweden | 1 | <1% |
Israel | 1 | <1% |
Switzerland | 1 | <1% |
Austria | 1 | <1% |
Other | 3 | 1% |
Unknown | 276 | 93% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 82 | 28% |
Researcher | 75 | 25% |
Student > Master | 35 | 12% |
Student > Bachelor | 26 | 9% |
Other | 15 | 5% |
Other | 33 | 11% |
Unknown | 31 | 10% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 121 | 41% |
Biochemistry, Genetics and Molecular Biology | 75 | 25% |
Engineering | 16 | 5% |
Neuroscience | 12 | 4% |
Chemistry | 9 | 3% |
Other | 29 | 10% |
Unknown | 35 | 12% |