Detection of Individual RNA in Fixed Cells and Tissues by Chromogenic ISH

Meng Jiang; Chen Lin; Rongqin Ke

doi:10.21769/BioProtoc.3510

Detection of Individual RNA in Fixed Cells and Tissues by Chromogenic ISH

Bio Protoc. 2020 Feb 5;10(3):e3510. doi: 10.21769/BioProtoc.3510.

Authors

Meng Jiang¹, Chen Lin¹, Rongqin Ke¹

Affiliation

¹ Center for Precision Medicine, School of Biomedical Sciences and School of Medicine, Huaqiao University, Quanzhou, Fujian, 362021, China.

Abstract

Visualization of RNA molecules in situ helps to better understand the functions of expressed genes. Currently, most conventional in situ hybridization methods for visualization of individual RNAs are based on fluorescence detection. Herein we present a chromogenic in situ hybridization protocol for visualization of single RNA molecules in fixed cells and tissues. The protocol is based on padlock probing and rolling circle amplification to generate detectable chromogenic signal from single RNA molecules. Chromogenic signal can avoid background autofluorescence and can be preserved for a longer period than fluorescence signal.

Keywords: Chromogenic in situ hybridization; Gene expression; Padlock probe; Rolling circle amplification.