Detection of Apoptosis by a Modified Trichrome Technique

This report describes a cytological method that allows the easy detection and quantification of apoptotic cells and bodies. By simple and inexpensive modifications of Crossmon's trichrome method, that consist of omitting the stain in Mayer acid hemalum, reduction of the staining time and concentration of fuchsin and orange G, and increasing the stain of light green to 30 min, differential staining of apoptotic and normal nuclei in paraffin sections was obtained. The procedure was applied to the study of the apoptotic process evoked by cyclophosphamide in the thymus of mice, Peyer's patches, and crypts of the small intestine. Quantitative comparison of this method with H&E demonstrated statistically no significant differences. The assay is easy and reliable, and its application to cell biology and experimental pathology offers many possibilities. (The J Histotechnol 22:301, 1999)