Isolation of cardiac conduction system of rat heart by laser capture microdissection

Objective:To isolate cells of cardiac conduction system (CCS) with laser capture microdissection (LCM) and extract and evaluate quality of small amount of RNA from cells of CCS. Methods: Cryostat sections were followed by H-E staining. 20 pieces of H-E stained cryostat sections were scraped and its RNA was assessed to insure that RNA didn’t degrade in dyeing and dehydration process. Cells of CCS were captured with LCM and quality of small amount of RNA was verified with RT-PCR.Results: Cells of CCS isolated with LCM had clear morphology after staining. High quality RNA was extracted from LCM samples and scraped tissues; 18S rRNA and 28S rRNA were seen distinctly on gel electrophoresis. Low level of small amount of RNA extracted from LCM sample was below the limit of detection on gel electrophoresis or ultraviolet spectrophotometer. The housekeeping genes β-actin and GAPDH were successfully amplified with small amount of RNA.Conclusion:This study resolves the problem of acquiring material of CCS precisely that hinders gene research of CCS. It is found out that the method is easy and reliable to extract and assess the quality of small amount of RNA from microdissected cells of CCS.