FFPE Tissue Pretreatment for RNA CISH: A Procedure to Process Formalin-Fixed Paraffin-Embedded Tissue Sections for RNA Chromogenic In Situ Hybridization

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 demonstrates the steps involved in the pretreatment of FFPE tissue specimens for RNA chromogenic in situ hybridization. The pretreatment process helps maintain nucleic acid integrity within the cells and overall tissue morphology.

Protocol

All procedures involving human participants have been performed in compliance with the institutional, national and international guidelines for human welfare and have been reviewed by the local institutional review board.

1. Preparation of the Materials

1. Preparation of 1x target retrieval reagent
   1. In a large beaker, add 70 mL of 10x target retrieval reagent (see the Table of Materials) to 630 mL of distilled water.
   2. Place the beaker on a heating plate with a magnetic stirrer. Cover it with aluminum foil.
   3. Boil its contents at 100 °C for 10....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
HybEZ Oven (110v)	Advanced Cell Diagnostics Inc.	321710
HybEZ slide rack	Advanced Cell Diagnostics Inc.	300104
ImmEdge Hydrophobic Barrier Pen	Advanced Cell Diagnostics Inc.	310018
RNAscope H202 & Protease Plus Reagent	Advanced Cell Diagnostics Inc.	322330	Hydrogen Peroxyde x2 and Protease Plus x 1
RNAscope Target Retrieval Reagents	Advanced Cell Diagnostics Inc.	322000