Comparison and optimization of DNA recovery from sperm vs. epithelial cells using laser capture microdissection technology and an immunofluorescent staining system

Abstract Laser capture microdissection (LCM) is an accurate and robust tool for isolation and separation of pure cell populations from heterogeneous mixtures via direct visualization and collection of elements. In this study, the capability of the P.A.L.M. ® LCM for separation of sperm and epithelial cells, combined with immunofluorescent Sperm HY-LITER™ stain was assessed. LCM was evaluated for its sensitivity, specificity, and reproducibility using sperm–epithelial mixture slides with varying sperm counts and ages ranging 1–9 years. In addition, slides stained with immunofluorescent dyes were tested. Separation of sperm and epithelial cells using LCM is a robust method for processing of mixed samples and generates interpretable single source DNA profiles. Immunofluorescent staining method is compatible with the platform.