A full color system for quantitative assessment of histochemical and immunohistochemical staining patterns

Quantitative measures of staining distributions are important to compare the presence and patterns of cells or macromolecules. Typically, achromatic thresholding systems are used to compare staining distributions, Achromatic video signals, however, lack sufficient resolution to identify and compare chromatic changes. The purpose of this study is to describe a full color system for analysis of chromatic staining distributions. The hardware system includes a Leitz Diaplan microscope, video camera, GVP videoboard and Amiga 3000 computer. Software was developed in "C" to partition the video signal into hue (H), saturation (S) and value (V). Also, percentage of stained area was determined. Kodak color filters were used to assess the accuracy and precision of the system. Craniofacial tissues were stained with varying concentrations of toluidine blue and primary anti-BrdU antibodies. HSV and the percentage of stained areas were determined and displayed low coefficients of error. HSV values also performed as expected for standard filters as well as cellular staining concentrations. This system is easily implemented and should be useful for comparing chromatic changes with any color resulting from histochemical or immunohistochemical procedures.