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

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 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 Preparation of 1x target retrieval reagent In a large beaker, add 70 mL of 10x target retrieval reagent (see the Table of Materials) to 630 mL of distilled water.</…

Divulgaciones

The authors have nothing to disclose.

Materials

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

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FFPE Tissue Pretreatment for RNA CISH: A Procedure to Process Formalin-Fixed Paraffin-Embedded Tissue Sections for RNA Chromogenic In Situ Hybridization. J. Vis. Exp. (Pending Publication), e20473, doi: (2023).

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