A Selective Stain for Mitotic Figures, Particularly in the Developing Brain

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Abstract
A selective stain for mitotic figures is valuable where autoradiographic counting is not required, especially in the developing brain. Most work in this field was based on conventional nuclear stains which do not differentiate mitotic figures from resting cells by color. Hematoxylin, Feulgen, gallocyanin and Nissl methods were used particularly. The method described uses a modified Bouin fixative, followed by hydrolysis in 1 N HCl. Mitotic figures are selectively stained using crystal violet, with nuclear fast red as the counterstain for resting cells. The method was tested using material from postnatal and fetal sheep, guinea pigs and rats. Using paraffin-mounted serial sections it is applicable to all organs. The method was very successful on developing rat brain, paticularly for detail and quantitative estimation in the early stages of prenatal development, which was of primary interest. Nucleated cells of the erythrocytic series, keratin and what appeared to be mast cells were found to stain. When nuclear counting or cell recognition were required these did not cause any difficulty, except in prenatal liver. The highly selective method presented stains mitotic figures, in all tissue tested, an intense blue against a background of red resting cells.