Histological staining methods preparatory to laser capture microdissection significantly affect the integrity of the cellular RNA.

BMC Genomics
Hongyang WangJ Frederic Mushinski

Abstract

Gene expression profiling by microarray analysis of cells enriched by laser capture microdissection (LCM) faces several technical challenges. Frozen sections yield higher quality RNA than paraffin-imbedded sections, but even with frozen sections, the staining methods used for histological identification of cells of interest could still damage the mRNA in the cells. To study the contribution of staining methods to degradation of results from gene expression profiling of LCM samples, we subjected pellets of the mouse plasma cell tumor cell line TEPC 1165 to direct RNA extraction and to parallel frozen sectioning for LCM and subsequent RNA extraction. We used microarray hybridization analysis to compare gene expression profiles of RNA from cell pellets with gene expression profiles of RNA from frozen sections that had been stained with hematoxylin and eosin (H&E), Nissl Stain (NS), and for immunofluorescence (IF) as well as with the plasma cell-revealing methyl green pyronin (MGP) stain. All RNAs were amplified with two rounds of T7-based in vitro transcription and analyzed by two-color expression analysis on 10-K cDNA microarrays. The MGP-stained samples showed the least introduction of mRNA loss, followed by H&E and immunofluore...Continue Reading

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Citations

Nov 8, 2007·PloS One·Konstantin KhodosevichHannah Monyer
Apr 11, 2009·World Journal of Gastroenterology : WJG·Stephan GehringStephen H Gregory
May 14, 2016·Scandinavian Journal of Clinical and Laboratory Investigation·Lea LandoltOystein Solberg Eikrem
Jan 18, 2016·Journal of Microbiological Methods·Oleg V Podgorny, Vassili N Lazarev
Dec 15, 2010·Biomarker Insights·Marc A Williams
Jan 18, 2018·Nature Communications·A GanguliR Bashir
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Jan 4, 2013·Physiological Genomics·Jerry WeiPeter Williamson
Nov 7, 2019·The American Journal of Forensic Medicine and Pathology·Papanu Suttipasit
May 7, 2020·Histochemistry and Cell Biology·Alexandra PoteAna Marques-Ramos
Aug 25, 2019·Medicina·Katiane de Almeida da CostaCarla de Azevedo Piccinato
Jun 1, 2017·Histochemistry and Cell Biology·P Garrido-GilJose L Labandeira-Garcia
Apr 13, 2012·Applied Immunohistochemistry & Molecular Morphology : AIMM·Liang ChengRodolfo Montironi

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Methods Mentioned

BETA
chip
in vitro transcription
electrophoresis
chips
PCR
laser capture microdissection

Software Mentioned

GenePixPro

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