RNA CISH Signal Detection: A Technique to Detect Chromogenic Signals During RNA In Situ Hybridization in Intact Tissue Specimens

Published: April 30, 2023

Abstract

Source: Outh-Gauer, S. et al. Chromogenic In Situ Hybridization as a Tool for HPV-Related Head and Neck Cancer Diagnosis. J. Vis. Exp. (2019)

This video presents the protocol for signal detection during RNA chromogenic in situ hybridization. This technique can be used to diagnose an active oncogenic infection visually in an intact tissue specimen.

Protocol

1. Preparation of Counterstaining Reagents Prepare 50% hematoxylin. In a fume hood, add 100 mL of Gill’s hematoxylin I (see the Table of Materials) to 100 mL of distilled water in a staining dish. NOTE: The 50% hematoxylin staining solution can be reused for up to 1 week. Prepare 0.02% (w/v) ammonia water (bluing reagent). In the fume hood, add 1.43 mL of 1 N ammonium hydroxide to 250 mL of dist…

Divulgaciones

The authors have nothing to disclose.

Materials

Hematoxylin solution, Gill No. 1   Merck GHS132
RNAscope 2.5 HD Detection Reagents-BROWN  Advanced Cell Diagnostics Inc.  322310 This kit includes amplification reagents AMP1, AMP2, AMP3, AMP4, AMP5 and AMP6, and detection reagents DAB-A and DAB-B
RNAscope Probe- HPV16/18  Advanced Cell Diagnostics Inc. 311121
HybEZ slide rack   Advanced Cell Diagnostics Inc. 300104
ImmEdge Hydrophobic Barrier Pen Advanced Cell Diagnostics Inc.  310018

Tags

Play Video

Citar este artículo
RNA CISH Signal Detection: A Technique to Detect Chromogenic Signals During RNA In Situ Hybridization in Intact Tissue Specimens. J. Vis. Exp. (Pending Publication), e20475, doi: (2023).

View Video