Skip to Main content Skip to Navigation
Journal articles

smiFISH and FISH-quant - a flexible single RNA detection approach with super-resolution capability.

Résumé : Single molecule FISH (smFISH) allows studying transcription and RNA localization by imaging individual mRNAs in single cells. We present smiFISH (single molecule inexpensive FISH), an easy to use and flexible RNA visualization and quantification approach that uses unlabelled primary probes and a fluorescently labelled secondary detector oligonucleotide. The gene-specific probes are unlabelled and can therefore be synthesized at low cost, thus allowing to use more probes per mRNA resulting in a substantial increase in detection efficiency. smiFISH is also flexible since differently labelled secondary detector probes can be used with the same primary probes. We demonstrate that this flexibility allows multicolor labelling without the need to synthesize new probe sets. We further demonstrate that the use of a specific acrydite detector oligonucleotide allows smiFISH to be combined with expansion microscopy, enabling the resolution of transcripts in 3D below the diffraction limit on a standard microscope. Lastly, we provide improved, fully automated software tools from probe-design to quantitative analysis of smFISH images. In short, we provide a complete workflow to obtain automatically counts of individual RNA molecules in single cells.
Complete list of metadatas

Cited literature [27 references]  Display  Hide  Download

https://hal-pasteur.archives-ouvertes.fr/pasteur-01375913
Contributor : Florian Mueller <>
Submitted on : Monday, October 3, 2016 - 5:46:05 PM
Last modification on : Thursday, September 24, 2020 - 5:06:02 PM
Long-term archiving on: : Friday, February 3, 2017 - 2:07:22 PM

File

2016_Tsanov_NAR_smiFISH.pdf
Publication funded by an institution

Licence


Distributed under a Creative Commons Attribution - NonCommercial 4.0 International License

Identifiers

Citation

Nikolay Tsanov, Aubin Samacoits, Racha Chouaib, Abdel-Meneem Traboulsi, Thierry Gostan, et al.. smiFISH and FISH-quant - a flexible single RNA detection approach with super-resolution capability.. Nucleic Acids Research, Oxford University Press, 2016, ⟨10.1093/nar/gkw784⟩. ⟨pasteur-01375913⟩

Share

Metrics

Record views

1816

Files downloads

1138