Visualization of RNA at the Single Cell Level by Fluorescent in situ Hybridization Coupled to Flow Cytometry

Alice Coillard; Elodie Segura

doi:10.21769/BioProtoc.2892

Visualization of RNA at the Single Cell Level by Fluorescent in situ Hybridization Coupled to Flow Cytometry

Bio Protoc. 2018 Jun 20;8(12):e2892. doi: 10.21769/BioProtoc.2892.

Authors

Alice Coillard¹, Elodie Segura¹

Affiliation

¹ Institut Curie, PSL Research University, INSERM U932, Paris, France.

Abstract

The protocol described here has been developed to detect RNA at the single cell level. Fluorescent probes hybridize to target RNAs and are detected by flow cytometry after multiple amplification steps. Different types of RNA can be detected such as mRNA, long noncoding RNA, viral RNA or telomere RNA and up to 4 different target probes can be used simultaneously. We used this protocol to specifically measure the expression of two transcription factor mRNAs, MAFB and IRF4, in human monocytes.

Keywords: Flow cytometry; Human; Hybridization; In situ; RNA; Single cell.