Principles and practical applications of laser capture microdissection

Laser capture microdissection (LCM) allows selective harvesting of specific cell types from paraffin-embedded or frozen sections. Recommendations for specimen preparation are presented, as well as rnicrodissection protocols. Precautions against cross-contamination of patients' samples are presented. Applications of this technology include analysis of genomic DNA, RNA and protein. Examples of types of analyses of DNA from LCM-harvested tissues include mutation analysis by sequencing, gene promoter methylation studies, comparative genomic hybridization, fluorescence in situ hybridization for gene copy number characterization and analyses of microsatellite instability and loss of heterozygosity. RNA isolated by LCM from fresh or frozen sections has been used for reverse transcriptase-PCR (real-time and regular) and amplified for microarray analysis.