1. Fix in absolute alcohol; imbed in celloidin; cut.
2. Stain in hæmatoxylin; differentiate in acid alcohol.
3. Wash in water.
4. Stain in filtered carmine mixture (carmine 1 grm., ammonium chlorate 2 grms., lithium carbonate O.5 grm., water 50 cc.; bring to boiling point, and when cool, add 20 cc. of strong liquid ammonia. Keep in dark; can be used after 2-3 days and gives good results up to 14 days) 2 parts, strong ammonia 3 parts, methyl alcohol 6 parts. Make fresh each time it is used, as it soon precipitates; do not filter; stain few sections at a time ¾-1 hour.
5. Differentiate 1-2 minutes in a mixture of absolute alcohol 4 parts, methyl alcohol 2 parts, water 5 parts.
6. Wash in 80 per cent alcohol.
7. Dehydrate in absolute alcohol.
8. Clear in xylol; mount in balsam.
Glycogen is stained red; nuclei blue; dense connective-tissue, mast-cell granules, protoplasm of gastric glands, etc., red; but these can all be distinguished morphologically from glycogen. This is by far the best method for the staining of glycogen.
XII. HYALIN. Epithelial hyalin (colloid) stains red or violet with hæmatoxylin and eosin; it takes the other acid dyes and stains to some degree with basic aniline stains. Van Gieson’s stains it a yellow, orange or brownish-pink. Kresyl-echt-violett gives it a deep indigo-blue color or a more green robin-egg blue. Connective-tissue hyalin stains deep brilliant red with Van Gieson’s; this is the best method for differentiating connective-tissue hyalin from amyloid or epithelial hyalin. Russell’s method also stains hyalin red.