

OTHER SPECIAL STAINS 117 



solutions from Griibler ready for use ; the powder for Irishman's stain 

 may be obtained from the same source, and the solution made up by 

 the worker himself. Cabot states that Wright's stain can be obtained 

 from the Harvard Co-operative Society, Boylston Street, Boston, U.S.A. 



Neisser's Stain. (a) The following is the original method introduced 

 by Neisser as an aid to the diagnosis of the diphtheria bacillus. Two 

 solutions are used as follows : (a) 1 grm. methylene-blue (Griibler) is 

 d'ssolved in 20 c.c. of 96 per cent, alcohol, and to the solution are added 

 950 c.c. of distilled water and 50 c.c. of glacial acetic acid ; (b} 2 grms. 

 Bismarck-brown (vesuvin) dissolved in a litre of distilled water. Films 

 are stained in (a) for 1-3 seconds or a little longer, washed in water, 

 stained for 3-5 seconds in (ft), dried, and mounted. The protoplasm of 

 the diphtheria bacillus is stained a faint brown colour, the granules a blue 

 colour. Neisser considers that this reaction is characteristic of the 

 organism, provided that cultures on Loffler's serum are used and examined 

 after 9-24 hours' incubation at 34 to 35 C. Satisfactory results are not 

 always obtained in the case of films prepared from membrane, etc., but 

 there is no doubt that here also the method is one of considerable 

 value. 



(b) The following is Neisser's modified cresoidin method : 



1. Stain films for a few seconds in a mixture of solutions A and B, 



two parts of the former to one of the latter. 



A. Methylene-blue .... 1 part. 

 Absolute alcohol .... 50 parts. 

 Glacial acetic acid . . . 50 ,, 

 Distilled water .... 1000 ,, 



B. Crystal-violet (Hb'chst) . . 1 part. 

 Absolute alcohol . . . 10 parts. 

 Distilled water .... 300 , , 



2. Wash in water, and 



3. Stain in cresoidin solution (1 : 300) for a few seconds (the cresoidin 



should be dissolved in warm water and the solution then 

 filtered). 



4. Wash in water, dry, and mount. 



Sabouraud's Method for Staining Trichophyta. Remove the fat from 

 the hair or epithelial squames with chloroform. Place in a test-tube 

 with 10 per cent, formol, and warm for two or three minutes till ebullition 

 commences. Wash well in distilled water, and stain for one minute in 

 Sahli's blue, which is made up as follows : 



Distilled water ...... 40 parts. 



Saturated watery methylene-blue . . . 24 ,, 

 5 per cent, solution of borax in water . 16 ,, 



Mix the constituents. Allow to stand for a day, and filter. After 

 staining, wash in water, dehydrate with absolute alcohol, clear in xylol, 

 and mount in balsam. 



