Pathology Image Analysis (PIA)
Applications
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| Nuclear, cytoplasmic, membranous |
Tissue microarray | Microvessel density | Nuclear cell counting | Ploidy analysis |
Rare event detection | |
Scoring of nuclear, cytoplasmic and membranous staining
(pixel-based analysis)
- Scan at low or high magnification
- For IHC set color thresholds to define pixels that correspond to chromogen (brown threshold) and counterstain (blue threshold) that can be checked using overlay tool and modified as necessary (Tretiakova M et al, J Clin Pathol, 2006).
- Select representative region areas for analysis which can be standard size, free-form, or whole tissue area
- Get standard measurements: brown intensity, region area, tissue area, blue area, brown area, brown/blue ratio (% positive) etc
- Compute integrated optical density (IOD), which is directly proportional to the concentration of molecule recognized by the stain according to Beer-Lambert Law (Oberholzer et al, 1996). IOD is a proxy for antigen content and it's calculated as intensity multiplied by brown area in microns
- For H&E and other stains thresholds are set to analyze separately nuclear compartments (i.e. blue), cytoplasms and fibrous tissues (pink), areas of hemorrhage (red), necrosis or other tissue elements (Fig. Rodent mammary gland)
