A rapid and simple staining method, using Toluidine Blue, for analysing mitotic figures in tissue sections |
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Authors: | Pasquale Chieco Maura Pagnoni Eleonora Romagnoli and Cinzia Melchiorri |
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Institution: | (1) Institute of Oncology, Viale Ercolani 4, I-40138 Bologna, Italy;(2) Second Department of Obstetrics and Gynaecology, University of Bologna, Bologna, Italy |
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Abstract: | Summary Mitotic index is a clinically important parameter in cancer pathology. We developed a staining method using Toluidine Blue
to detect efficiently and rapidly mitotic figures in sections of formalin-fixed paraffin-embedded human and rat tisues. Sections
were stained at acid pH with a 0.01% Toluidine Blue solution after removal of RNA with hydrochloric acid or ribonuclease.
The optimal pH of the TB staining solution was found to be 4.5 for rat tissues and 3.5 for human tissues. This procedure stained
mitotic figures much more intensely than other (extra)cellular structures. A quantitative estimate of the total number of
nuclei in the field where mitotic figures were counted, was obtained in an adjacent section hydrolysed in 5 N hydrochloric
acid and stained by the Feulgen reaction with a Schiff-type reagent containing 0.01% Toluidine Blue. This method specifically
stained interphase and mitotic nuclei and the field cellularity could be quantified by image cytometry. When these procedures
were performed on two consecutive serial sections, a mitotic index could be determined accurately by relating the count of
mitotic figures to the number of tumour cells. |
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