Rapid Loss of RNA Detection by in Situ Hybridization in Stored Tissue Blocks and Preservation by Cold Storage of Unstained Slides

Javier A. Baena-Del Valle, Qizhi Zheng, Jessica L. Hicks, Helen Fedor, Bruce J. Trock, Colm Morrissey, Eva Corey, Toby C Cornish, Karen S. Sfanos, Angelo M. De Marzo

Research output: Contribution to journalArticle

Abstract

Objectives Recent commercialization of methods for in situ hybridization using Z-pair probe/branched DNA amplification has led to increasing adoption of this technology for interrogating RNA expression in formalin-fixed, paraffin-embedded (FFPE) tissues. Current practice for FFPE block storage is to maintain them at room temperature, often for many years. Methods To examine the effects of block storage time on FFPE tissues using a number of RNA in situ probes with the Advanced Cellular Diagnostic's RNAscope assay. Results We report marked reductions in signals after 5 years and significant reductions often after 1 year. Furthermore, storing unstained slides cut from recent cases (<1 year old) at -20°C can preserve hybridization signals significantly better than storing the blocks at room temperature and cutting the slides fresh when needed. Conclusions We submit that the standard practice of storing FFPE tissue blocks at room temperature should be reevaluated to better preserve RNA for in situ hybridization.

Original languageEnglish (US)
Pages (from-to)398-415
Number of pages18
JournalAmerican Journal of Clinical Pathology
Volume148
Issue number5
DOIs
StatePublished - Nov 1 2017

Keywords

  • Branched DNA amplification
  • Formalin-fixed
  • paraffin-embedded tissues
  • RNA in situ hybridization

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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