Traditional histopathology analysis has two basic problems. First, it isn't granular enough: Pathologists typically grade overall marker-staining intensity using a four-point scale. The other problem is that these measurements don't account for the sometimes-subtle changes in subcellular localization that can indicate disease. Beta-catenin, for instance, is a biomarker for colon cancer, but only when localized to the nucleus. Yale University pathologists Bob Camp and David Rimm developed a series of algorithms called AQUA (Automated Quantitative Analysis) to overcome these shortcomings, providing compartment-specific staining-intensity data over the range of 0 to 255, which has been shown to be directly proportional to absolute protein concentrations.
In this example, estrogen receptor staining is being quantified in cancerous breast tissue. The cells are stained with three tags: cytokeratin (green, A), which differentiates the tumor from surrounding tissue; DAPI (blue, B), which stains nuclei; and an anti-estrogen (ER) receptor antibody (red, C).
Cytokeratin staining is ...
