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 |
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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).