The current use of tumour microvascular density measurements in clinical practice is limited by the methods employed to identify and quantify the microvessels. The aim of this study was to assess tumour vascularity with three of the most commonly used endothelial markers and with different pre-treatment methods to determine which combination results in the best visualisation of endothelial cells. Serial sections of colorectal carcinoma were treated with microwave oven irradiation, pressure cooking or proteolytic digestion prior to immunostaining with von Willebrand factor (vWf), CD31 or CD34. The sensitivity of staining was enhanced by microwave pre-treatment for CD31 and CD34, and trypsinization for vWf. There was a statistically significant difference between microvascular counts obtained using antibodies for CD34 and vWf (p=0.004), and CD34 and CD31 (p=0.007), but not for CD31 and vWf (p=0.775). We conclude that the use of CD34 antibody on microwaved tissues is the most sensitive method to visualize and quantify vascular density in tumours.

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