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Laser Capture Microdissection of Paraformaldehyde-Fixed Mouse Liver Tissue for RNA Analysis
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Laser capture microdissection for analysis of gene expression in formalin-fixed paraffin-embedded tissue
Ru Jiang1, Rona S Scott, Lindsey M Hutt-Fletcher
1Department of Microbiology and Immunology, Center for Molecular and Tumor Virology and Feist-Weiller Cancer Center, Louisiana State University Health Sciences Center, Shreveport, LA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|July 16, 2011
Summary
Analyze gene expression in archived tissues using laser capture microdissection and reverse transcriptase quantitative PCR. This method quantifies relative gene expression levels in individual cells from preserved tissue specimens.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Archived tissue specimens are valuable resources for retrospective studies.
- Analyzing gene expression in specific cells within these tissues presents technical challenges.
Purpose of the Study:
- To describe standard methodologies for analyzing relative gene expression.
- To enable gene expression analysis in individual cells from formalin-fixed paraffin-embedded tissues.
Main Methods:
- Laser capture microdissection (LCM) for isolating specific cells.
- Reverse transcriptase real-time quantitative PCR (RT-qPCR) for gene expression analysis.
Main Results:
- Successful application of LCM combined with RT-qPCR for gene expression analysis.
- Demonstration of a powerful tool for analyzing archived tissue specimens.
Conclusions:
- This combined technique provides a robust method for studying gene expression in historical samples.
- Enables detailed molecular analysis of individual cells within preserved tissues.
