Gene expression analysis requires a sound basis of cell material to obtain realistic results. Tissue, however, consists of diverse types of cells, which often differentially express target genes, so that cell populations need to be selected. If tissue diversity is moderate and negligible, manual microdissection can be the cost-efficient method of choice. In contrast, the advantage of laser microdissection is a very exact selection down to the level of a single cell, but often with a considerable time needed to get enough material for the following analyses. The latter issue and the method of tissue preparation needed for laser microdissection are the main problems to solve if RNA, highly sensitive to degradation, shall be analyzed. This method focuses on optimized laser microdissection procedures for RNA analysis, drawing on the very heterogeneous tissue of prostatic adenocarcinoma.