Laser capture microdissection combined with functional grouping cDNA microarray analysis in cerebral ischemia / 西安交通大学学报(医学版)

ABSTRACT

Objective To develop a method to analyze functional grouping cDNA microarray gene expression in microvessels and neurons from rat brain using laser capture microdissection(LCM). Methods Microvessels and neurons were captured using the PixCell ⅡLCM instrument. The total RNA was extracted from the LCM samples according to the manufacturer's protocol of RNAqueous-Micro kit. The total RNA were processed for one round of T7-based RNA amplification; cDNA probe were synthesized using gene-specific primer and labeled for cDNA microarray analysis. Results Amplified RNA from microvessels and neurons allowed us to measure 96 gene expression in functional grouping cDNA microarray. Conclusion LCM allowed us to harvest pure microvessels and neurons from their native tissue environment, combined with methods of T7 RNA amplification and gene-specific primer amplification so that we can analyze functional grouping cDNA profiling in LCM samples.