XVII. MITOTIC FIGURES. Various histologic methods devised for the study of mitoses can be applied to the demonstration of these in neoplasms, inflammation and regeneration. Flemming’s solution or mercuric chloride fixation gives best results, although formol, or even absolute alcohol, when used quickly and carefully gives fair results if tissue is very fresh.
1. Flemming’s Solution and Safranin.
1. Fix small pieces of fresh tissue in Flemming’s, in the dark, for 24 hours; wash 24 hours; after-harden in graded alcohols; imbed and cut.
2. Stain in 1 per cent water solution or saturated aniline water solution of safranin, or 1 per cent water methyl violet for 12-24 hours, or carbol-fuchsin for one hour.
3. Differentiate quickly in a 0.5-0.0001 HCl in 70 per cent alcohol and then in absolute alcohol until stain no longer comes away in clouds and nuclei have right shade.
4. Clear in xylol; mount in balsam.
Fat is black; mitoses stand out sharply; tubercle-bacilli may be stained black or red.
2. Fixation in mercuric-chloride may be followed by Ehrlich-Biondi-Heidenhain’s stain (saturated aqueous orange 100 cc., saturated aqueous acid fuchsin 20 cc., saturated aqueous methyl green 50 cc.) 12 grms. of Grübler’s prepared stain dissolved in 100 cc. of distilled water, for stock solution. For staining take 1 cc. of stock solution, water 30 cc., ½ per cent watery acid fuchsin 3 cc., and 2 per cent acetic 5-6 drops. Stain 2-24 hours; wash in 90 per cent alcohol; dehydrate in absolute; clear in xylol; balsam.
Resting nuclei are bluish; mitoses and fragments of leukocyte nuclei dark green; red blood cells orange red; protoplasm and connective-tissue fuchsin red.
3. Benda’s Iron-Haematoxylin Method.