Microvessel density counting in breast cancer. Slides vs. digital images.
OBJECTIVE
To develop a program to assist the pathologist in the acquisition and evaluation of digital images to determine microvessel density (MVD) in tissues.


METHODS
Ten cases of breast cancer with a high degree of neovascularization were selected. A standard immunohistochemical method was used to highlight the microvessels (monoclonal anti-factor VIII, avidinbiotin-peroxidase complex method). Two pathologists (one senior [S] and one junior [J]) evaluated four areas of highest neovascularization ("hot spots") in the tumors. Microscopically MVD was determined in four chosen areas (400:1). From the center of each area two digital images were acquired at a magnification of 200:1. All counts made by microscopic observation were compared with those made on the digital images. To compare MVD counting at different resolution, two sets of images at different sampling densities (320 x 240 and 1,600 x 1,200) were assessed by the two pathologists.


RESULTS
We obtained a good correlation (r = .98 for S and .96 for J) between the MVD counts obtained at the microscope (192.8 MV/mm2 [mean of S] and 181.8 MV/mm2 [mean of J]) and the MVD counts from digital images (153.2 MV/mm2 [mean of S] and 171.0 MV/mm2 [mean of J]) at high resolution. The counts were lower for digital images at lower sampling density (125.0 MV/mm2 [mean of S] and 78.2 MV/mm2 [mean of J]). With low-resolution digital images only S maintained a good correlation (r = .96 for S and .34 for J) with the microscopic evaluation of MVD. Interobserver analysis showed a good correlation (r = .82 for the microscope and r = .78 for the digital images) of MVD evaluated either at the microscope or in high-resolution digital images.


CONCLUSIONS
We demonstrated the functionality and usefulness of our program in performing MVD evaluation. Considering the capabilities of the program to store all images and microvessel marks and the reliability of MVD evaluation based on digital images, we consider this program the first step toward fully automated MVD assessment.
Institute of Molecular Pathology and Immunology, University of Porto, Porto, Rua do Dr. Roberto Frias s/n, 4200 Portugal. dcruz@ipatimup.pt
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Cruz
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Valentí
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Dias
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Seixas
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Schmitt
D001943:Breast Neoplasms / Q000098:blood supply* / Q000473:pathology; D003936:Diagnosis, Computer-Assisted; D006801:Humans; D019044:Image Cytometry; D007150:Immunohistochemistry; D008833:Microcirculation / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000473:pathology*; D015588:Observer Variation