6. When the background is pink, dehydrate quickly with absolute alcohol, or in 95 per cent by blotting on slide with xylol, until clear.
7. Clear in xylol.
8. Mount in balsam.
g. Van Gieson’s Method.
1. Stain in Weigert’s iron-hæmatoxylin, or overstain if any other hæmatoxylin is used. (Weigert’s gives the best results, as it does not decolorize so readily.)
2. Wash thoroughly.
3. Stain in Van Gieson’s mixture (acid fuchsin 1.5 grms., saturated water solution of picric acid [0.6 per cent] 150 cc. This mixture keeps well. Add 1 cc. of this stock solution to 10 cc. of saturated water solution of picric acid. Stain in this for 10 seconds).
4. Wash quickly; dehydrate in alcohol; clear in xylol or carbol-xylol; mount in balsam.
I have obtained the best results by making the Van Gieson’s mixture by taking an ordinary small staining-dish nearly full of saturated water solution of picric acid, and adding to this, drop by drop, sufficient saturated water solution of acid fuchsin to make the solution just dark enough so that the finger cannot be seen through the staining-dish. The hæmatoxylin-stained section is put into this mixture for a few seconds, until it appears to become lighter. The section is then washed in 95 per cent alcohol, dehydrated, cleared and mounted.
The Van Gieson method is extremely valuable in pathologic work, because of its varied differential reactions. The nuclei are brown or black, protoplasm is ochre-yellow, connective-tissue light red, voluntary and involuntary muscle yellow, axis-cylinders red, connective-tissue hyalin deep rose-red, epithelial hyalin yellow, orange or brownish, amyloid yellow or brownish pink, mucin yellow or brownish, fibrin yellow or brown, necrotic areas yellow or brownish, lime-salts brown to brownish blue or violet.