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The walls of angiogenic blood vessel capillaries are composed of two principal cell types, blood vessel endothelial cells ( BEC) and pericytes ( PC), whereas the walls of lymphatic capillaries are composed of lymphatic endothelial cells ( LEC). In this investigation we describe a practical application of NIH ImageJ software for quantitative image analysis for pericytes and endothelial cells in prostate cancer. We used a tissue microarray that contained 49 tissue cores (normal prostate tissue or prostatic carcinomas with Gleason scores of 6 through 10). These prostate cancer samples represented AJCC prognostic stages II, III, and IV. Slides were immunostained with anti- PDGFR-β antibody for identification of PC, and quantified as microvascular pericyte density ( MVPD); they were also immunostained with anti- CD34 antibody for identification of LEC and BEC simultaneously, and quantified as microvascular endothelial density ( MVED). CD31 and D2-40 immunostains were used to quantify BEC and lymphatic endothelial cells, respectively. Our results showed higher MVPD and MVED in prostate cancers with higher Gleason scores and higher stages, suggesting the prognostic utility of vascular image analysis in prostate pathology. This investigation demonstrates the feasibility, versatility, and ease of use of ImageJ software and pericyte-specific and endothelial-specific immunohistochemistry for quantitative image analysis in prostate pathology. [ABSTRACT FROM AUTHOR]