4. Stain on the slide or cover-glass in the aniline-methyl-violet (or gentian-violet) solution for 10 minutes. Wash off stain with physiologic salt solution.
5. Blot section with absorbent paper.
6. Cover section with Lugol’s (300-2-1) or a 5 per cent watery potassium iodide saturated with iodine. Leave on section for 1-5 minutes.
7. Blot off iodine.
8. Differentiate in aniline xylol (equal parts of xylol and aniline oil) until the nuclei become red.
9. Wash in xylol, blotting with absorbent paper. Repeat until section is transparent; then mount in balsam. All aniline oil must be removed before using the balsam.
Nuclei are red; fibrin deep blue; bacteria, mucin, keratin and Altmann’s granules also blue. The differentiation must be carefully controlled under the microscope, and should be stopped before the finest threads of fibrin begin to be decolorized.
XI. GLYCOGEN. Glycogen is soluble in water; and fixation and hardening must be carried out with absolute alcohol to prevent the solution of the glycogen. Tissue must be fixed immediately after death, as glycogen is quickly broken up. Its reaction with iodine is similar to that of amyloid, but it does not give the iodine-sulphuric-acid reaction that the latter substance does.
1. Best’s Iodine Method.
1. Fix and harden in absolute alcohol; imbed in paraffin; cut.