Rapid Loss of RNA Detection by In Situ Hybridization in Stored Tissue Blocks and Preservation by Cold Storage of Unstained Slides. Am J Clin Pathol 2017 Nov 02;148(5):398-415
Date
11/07/2017Pubmed ID
29106457Pubmed Central ID
PMC5848261DOI
10.1093/ajcp/aqx094Scopus ID
2-s2.0-85033584861 (requires institutional sign-in at Scopus site) 50 CitationsAbstract
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.
Author List
Baena-Del Valle JA, Zheng Q, Hicks JL, Fedor H, Trock BJ, Morrissey C, Corey E, Cornish TC, Sfanos KS, De Marzo AMAuthor
Toby Charles Cornish MD, PhD Professor in the Pathology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsAntigens, Neoplasm
Cold Temperature
Cryopreservation
Humans
In Situ Hybridization
Male
Mice
Paraffin Embedding
Prostatic Neoplasms
RNA
Tissue Array Analysis
Tissue Fixation
