Published online by Cambridge University Press: 14 March 2018
For over 5 years, here in Pathology at Tuiane, I have used color thresholding (CT) rather than monochrome thresholding (MT) for objective quantitation of histological, immunohistological, fluorescent and thick TEM sections. Technical aspects distinguishing CT from MT were recently described in detail, and this technical note represents an abbreviation of the last paper, which I recommend for those beginning to use video imaging technology. The system I use is the V150 (r) from ONCOR. Despite the advantages of today computers, data outcome, e.g., unbiased, still depends on good experimental design, strict control and repeatability. When the stain of interest is histochemically derived, my laboratory follows standard procedures found in the Handbook of Histopathological and Histochemical Techniques. When the stain is immunohistochemically derived, my laboratory checks cross reactivity of every antibody with known positive and negative control tissues besides using irrelevant antibodies or omitting the primary antibody. In addition, preabsorbing the antibody is done whenever its substrate is available.