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Sow FB, Gallup JM, Sacco RE, et al. Laser Capture Microdissection Revisited as a Tool for Transcriptomic Analysis: Application of an Excel-Based qPCR Preparation Software (PREXCEL-Q). Int J Biomed Sci. 2009;5(2):105-124
Sow, F. B., Gallup, J. M., Sacco, R. E., & Ackermann, M. R. (2009). Laser Capture Microdissection Revisited as a Tool for Transcriptomic Analysis: Application of an Excel-Based qPCR Preparation Software (PREXCEL-Q). International journal of biomedical science : IJBS, 5(2), 105-124.
Sow, Fatoumata B, et al. "Laser Capture Microdissection Revisited as a Tool for Transcriptomic Analysis: Application of an Excel-Based qPCR Preparation Software (PREXCEL-Q)." International journal of biomedical science : IJBS vol. 5,2 (2009): 105-124.
Sow FB, Gallup JM, Sacco RE, Ackermann MR. Laser Capture Microdissection Revisited as a Tool for Transcriptomic Analysis: Application of an Excel-Based qPCR Preparation Software (PREXCEL-Q). Int J Biomed Sci. 2009 Jun 01;5(2):105-124. PMID: 20556230; PMCID: PMC2885795.
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Int J Biomed Sci. 2009 Jun 01;5(2):105-124.

Laser Capture Microdissection Revisited as a Tool for Transcriptomic Analysis: Application of an Excel-Based qPCR Preparation Software (PREXCEL-Q).

International journal of biomedical science : IJBS

Fatoumata B Sow, Jack M Gallup, Randy E Sacco, Mark R Ackermann

Affiliations

  1. Department of Veterinary Pathology, College of Veterinary Medicine, Iowa State University, Ames, USA.

PMID: 20556230 PMCID: PMC2885795

Abstract

The ability to reliably analyze cellular and molecular profiles of normal or diseased tissues is frequently complicated by the inherent heterogeneous nature of tissues. Laser Capture Microdissection (LCM) is an innovative technique that allows the isolation and enrichment of pure subpopulations of cells from tissues under direct microscopic examination. Material obtained by LCM can be used for downstream assays including gene microarrays, western blotting, cDNA library generation and DNA genotyping. We describe a series of LCM protocols for cell collection, RNA extraction and qPCR gene expression analysis. Using reagents we helped develop commercially, we focus on two LCM approaches: laser cutting and laser capture. Reagent calculations have been pre-determined for 10 samples using the new PREXCEL-Q assay development and project management software. One can expect the entire procedure for laser cutting coupled to qPCR to take approximately 12.5-15 h, and laser capture coupled to qPCR to take approximately 13.5-17.5 h.

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