Whole-section imaging was performed using a NanoZoomer Digital Pathology slide scanner (Hamamatsu; Bridgewater, NJ) at the University of Washington Histology and Imaging Core in Seattle, WA. Slides were scanned in bright-field at a 20× objective and the digital images imported for analysis using Visiopharm software (Hoersholm, Denmark), which provides automated quantification of stained areas and uniform systematic random sampling on whole-section bright-field images (26 (link)). The software labeled the marker (HA, IαI, versican, and TSG-6)-positive area vs. tissue area using a project-specific configuration based on a threshold of pixel values. The relative and total tissue areas stained by the markers were measured. All the images were processed together using this configuration to generate the desired outputs (stained and unstained tissue areas measured in square micrometers).
Protocol detail
Automated Whole-Slide Quantification of Histological Markers
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Tissue
Versican
Corresponding Organization : Matrix Research (United States)
Other organizations : University of Manchester, Wellcome Centre for Cell-Matrix Research, Stanford University, Stanford Medicine
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Variable analysis
independent variables
- Whole-section imaging was performed using a NanoZoomer Digital Pathology slide scanner
- The relative and total tissue areas stained by the markers (HA, IαI, versican, and TSG-6) were measured
- Slides were scanned in bright-field at a 20× objective
- Visiopharm software was used for automated quantification of stained areas and uniform systematic random sampling on whole-section bright-field images
- None specified
- None specified
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