Laser Capture Microdissection for Analysis of Gene Expression in Formalin-Fixed Paraffin-Embedded Tissue
A combination of laser capture microdissection and reverse transcriptase real-time quantitative PCR provides a powerful tool for the analysis of relative gene expression in archived tissue specimens. This chapter describes standard methodologies that can be used to determine the relative levels of gene expression in individual cells captured from formalin-fixed paraffin-embedded tissues.
Key wordsFormalin-fixed tissues Laser capture microdissection Gene expression Reverse transcription Real-time quantitative PCR
This work was supported by grants DE 016669 and CA114416.
- 1.Curran, S., McKay, J. A., McLeod, H. L., Murray, G. I. (2000) Laser capture microscopy. Journal of Clinical Pathology 53, 64–68.Google Scholar
- 2.Fend, F., Emmert-Buck, M. R., Chuaqui, R., Cole, K., Lee, J., Liotta, L. A., Raffeld, M. (1999) Immuno-LCM: laser capture microdissection of immunostained frozen sections for mRNA analysis. American Journal of Pathology 154, 61–66.Google Scholar
- 3.Livak, K. J., Schmittgen, T. D. (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods 25, 402–408.Google Scholar
- 4.Pfaffl, M. W. (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Research 29, e45.Google Scholar