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Journal of Histochemistry and Cytochemistry, Vol. 51, 151-158, February 2003, Copyright © 2003, The Histochemical Society, Inc.

ARTICLE

Computerized Quantification of Tissue Vascularization Using High-resolution Slide Scanning of Whole Tumor Sections

Christophe F. Chantraina, Yves A. DeClercka, Susan Groshenb, and George McNamarac
a Division of Hematology–Oncology, Department of Pediatrics and Department of Biochemistry and Molecular Biology, Childrens Hospital Los Angeles and USC Keck School of Medicine
b Department of Preventive Medicine, USC Keck School of Medicine
c Congressman Dixon Cellular Imaging Core, Childrens Hospital Los Angeles Research Institute, Los Angeles, California

Correspondence to: Yves A. DeClerck, Div. of Hematology–Oncology, MS #54, Childrens Hospital Los Angeles, 4650 Sunset Boulevard, Los Angeles, CA 90027. E-mail: declerck@hsc.usc.edu

Assessment of tissue vascularization using immunohistochemical techniques for microvessel detection has been limited by difficulties in generating reproducible quantitative data. The distinction of individual blood vessels and the selection of microscopic fields to be analyzed remain two factors of subjectivity. In this study, we used imaging analysis software and a high-resolution slide scanner for measurement of CD31-immunostained endothelial area (EA) in whole sections of human neuroblastoma xenograft and murine mammary adenocarcinoma tumors. Imaging analysis software provided objective criteria for analysis of sections of different tumors. The use of the criteria on images of entire tumor section acquired with the slide scanner constituted a rapid method to quantify tumor vascularization. Compared with previously described methods, the "hot spot" and the "random fields" methods, EA measurements obtained with our "whole section scanning" method were more reproducible with 8.6% interobserver disagreement for the "whole section scanning" method vs 42.2% and 39.0% interobserver disagreement for the "hot spot" method and the "random fields," respectively. Microvessel density was also measured with the whole section scanning method and provided additional data on the distribution and the size of the blood vessels. Therefore, this method constitutes a time efficient and reproducible method for quantification of tumor vascularization.

(J Histochem Cytochem 51:151–158, 2003)

Key Words: angiogenesis, vascularization, endothelial area, CD31, immunohistochemistry, slide scanner, quantification


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