Qualitative cytological criteria for the validation of enzyme histochemical techniques.

Qualitative cytological criteria are concerned with the 'precision' of a histochemical reaction and form part of the general criteria for the validation of enzyme histochemical techniques. Three major problem areas are considered: (a) the general artifacts which interfere with the 'clean' appearance of the tissue sections; (b) the exact intracellular localization of the final reaction product and its relationship to specific subcellular compartments; (c) the problem of diffusion of enzymes or reaction products, or both, from their primary subcellular sites. Each of these points is discussed and illustrated by a few examples and the conclusion is drawn that by careful consideration of various criteria the enzyme cytochemical techniques can provide 'quantitative' information comparable to biochemical measurements of enzyme activity in tissue homogenates. In the example of localization of catalase with 3,3'-diaminobenzidine it is demonstrated that biochemical enzyme assays corrrelate closely with quantitative morphometric data obtained from cytochemical preparations through an automatic image analyser system.