MICROSCOPY 69 



general use. On p. 414, the authors list two methods for staining Actinomyces 

 in sections, although alum-hematoxylin followed by a strong eosin solution will 

 give good results, as will also the Gram method for paraffin sections, which is 

 as follows : Stain in anilin-methyl violet for 5-20 minutes ; wash in normal salt 

 solution or water; iodine solution (1:2:300) 1 minute; wash in water; absolute 

 alcohol, several changes, until no more color is given off and the section is ap- 

 parently decolorized; xylol; xylol and balsam. The so-called "clubs" do not 

 stain with Gram's stain while the central portion of the granule, the thin 

 filaments, do. 



Stains for Hair and Scrapings. — Adamson (1895) recommended clearing 

 with 5-10% KOH and staining by the Gram method. Chalmers and Marshall 

 (1914) suggest soaking scales in 40% KOH for some hours in a w^atch glass 

 in an incubator at 40° C. Transfer specimens to watch glass containing 15% 

 alcohol for 30 minutes, remove to slide, allow alcohol to evaporate, and dry over 

 flame; stain with anilin-gentian violet for 30 minutes. Treat with Gram's 

 iodine solution for 3 minutes ; decolorize with anilin oil for 30 minutes ; stain 

 in concentrated alcoholic eosin for 1 minute ; wash off eosin with anilin oil or 

 clove oil ; treat with xylol, and mount in balsam. 



Priestley (1917) recommends lactophenol (lactic acid 1 part, phenol 1 

 part, glycerol 2 parts, water 1 part) for clearing instead of 40% KOH; or 

 chloral hydrate crystals 2 parts, lactic acid 1 part, phenol crystals 1 part, may 

 be used. For staining he recommends treatment with chloroform to remove 

 the fat; boil for 2-3 minutes with formic acid; wash for a few minutes in 

 water, stain with Sahli 's methylene blue ; wash ; differentiate with alcohol, if 

 necessary; dehydrate, and mount in balsam. 



Bachman (1920) recommends the following procedure : Place scrapings 

 in a drop of water on a cover slip, tease thoroughly with a dissecting needle, 

 dry over a flame but do not scorch. Stain for 2 minutes; decolorize in 95% 

 alcohol for 15-30 seconds; immerse in distilled water 15-30 seconds; pour off 

 excess, dry by heat, and mount in balsam. The spores and mycelium will be 

 blue, the scrapings yellow. His dye is made as follows: saturated alcoholic 

 gentian violet 2.5 parts, distilled water 17.5 parts, orange G solution 9 parts, 

 acetic acid 1 part, 95% alcohol 5 parts. His orange G solution is orange G 

 2 parts, 95% alcohol 20 parts, water 80 parts. Decolorize with 10-20% KOH. 

 The host is not stained, the fungus appears yellowish red. 



Unna's method is to rub the scales of the epidermis in a little glacial 

 acetic acid between two slides. These are drawn apart and quickly dried 

 over a flame. The fat is removed by means of alcohol and ether, and the 

 preparations are stained in borax-methylene-blue. 



Instead of these slightly complicated methods, I have found that infected 

 hairs can be cleared sufficiently to show spores and mycelium by mounting in 

 a hydroxide solution, sodium or potassium, 10-30%. Sodium hydroxide is not 

 quite as satisfactory as potassium hydroxide and a 20% solution works with 

 sufficient rapidity to give good results without seriously macerating the hair. 

 The hair may be immersed in ether to dissolve off' oil or fat, and slight heating 



