Real time PCR from expression microdissection (xMD) of epithelial cells

The goal of this experiment was to determine the extent to which the xMD method can enrich for a specific cell type. Here we optimized each step of the xMD process from tissue retrieval, tissue transfer, film preparation, and RNA isolation to increase RNA yields and specficity. Duodenum (small intenstine) tissue was formalin fixed and processed to create routine FFPE slides. Matched RNA from a scrape of a unstained slide and AE1/AE3+ epithelial cells isolated by xMD were compared using TLDA qPCR microRNA array. FFPE slide samples were sectioned at 4um. The samples underwent xMD using AE1/AE3 antibody to select epithelial cells. The epithelial cell samples were matched to a scrape of the FFPE slide or bulk tissue.