EBVIEW — THOPICAL MEDICINE, ETC. 197 



Iron Hiematoxylin. Iron alum water solution 2 J to 4 per cent., six hours or longer. Staining— 

 Wash for about one minute in distilled water. Then in Heidenhain's Haematoxylin about continued 



twelve hours. Wash in water and differentiate under microscope in a watch-glass with 

 \ per cent, solution of iron alum. 



Belafield's Hsematoxylin. Must be used dilute about four or five drops to a Petri dish 

 of distilled water. Stain film in this for twenty-four to forty-eight houi-s. 



Borax Carmine. One part to about five or six parts of 70 per cent, alcohol. Differentiate 

 for about half to one hour in 2 per cent, acid (HCl) alcohol. 



After staining, films are passed through alcohol and xylol, and mounted in balsam. 



Films must never be allowed to dry during the whole process. 



Fixing in bulk. If fasces are solid, mix with saline and pour into test-tube to depth of 

 about one inch. Fill up with fixing fluid (as above, or Flemming's) and gently shake to mix 

 well. Allow fseces to settle ; this takes about one hour. Eemove supernatant fluid and 

 replace by water. Allow to settle and repeat, washing twice. Then add 70 per cent, 

 alcohol and iodine and leave for several hours. Eemove fluid and add stain — either dilute 

 Delafield's Hismatoxylin or Borax Carmine (after Borax Carmine add acid alcohol). Take 

 up into absolute alcohol by this sedimentation method. In another test-tube place at 

 bottom clove oil, then layer of mixture of clove oil and absolute alcohol, above this the faeces 

 in absolute alcohol. Faeces will gradually sink to the bottom, when fluid can be removed 

 and fresh clove oil added. Examine in clove oil. 



Plimmer'* has a new method for demonstrating trypanosomes so as to show the finest 

 cytological details. The following is the technique employed : — 



(1) Expose .1 cover-slip to the v.-ipour of osmic acid (1 per cent.), 1 c.c. glacial acetic acid 3-5 drops for 

 2 minutes; (2) place a drop of fresh blood on one corner of the slip and expose again to the vapour for 30 seconds; 

 (3) spread the film carefully and expose again for 15-30 seconds to the vapour until the surface appear no longer 

 moist ; (4) place slip in absolute alcohol for 10 minutes ; (5) immerse slip in faintly rose-coloured solution of per- 

 manganate of potash for 1 minute (2-3 drops of 1 per cent. sol. to 50 c.c. H^O) ; (6) wash in water for 5 minutes ; 

 (7) stain in the following modified Eomanowsky, made by mixing just before use — azur 1 (1 per cent.) 1 c.c. ; 

 eosin. Aj (1-1000) 2 c.c, H^O 8 c.c. for 15-30 minutes; (8) wash; (9) differentiate in orange tannin 

 30 seconds ; (12) alcohol xylol (2-3) two or three changes ; (13) xylol ; and mount. 



Instead of 7 to 13, any other method of staining can be used, according to what structures it is desired 

 particularly to show. 



Huisman^* studied 17 different methods for staining blood films, and found Jenner's best 

 and simplest, but he has improved upon it. 



The stain he uses is a mixture of equal parts of a 1-175 per cent, solution of solid azur- 

 blue in pure absolute methyl alcohol, and a 0-825 per cent, solution of eosin BA (Hochst) in 

 the same medium. He stains for two minutes without previously fixing the film. By this 

 method the nuclei appear a violet-blue, while the basophile protoplasm takes a light blue stain 

 which gives a good differentiation from the rest of the preparation. The red corpuscles are 

 rose-coloured. The neutrophile granules are rose-violet, violet, or violet-blue ; basophile 

 granules are blue ; the metachromatic basophile granules are violet-red, and the oxyphile 

 granules are red. The author also observes that some of the lymphocytes exhibit, when 

 stained by this formula, evidence of a fine blue or metachromatic granulation. 



Urtubey^* has invented what he calls a simple, rapid and certain method of preparing 

 the Leishman stain. Those who are accustomed to use the "soloids," and to be satisfied 

 with them, are not likely to resort to this new method, but a reference is given for any who 

 may wish to try it. It is simpler than the usual complicated process. A new Gram's 

 method, in which both the stain and the Gram's liquid are altered, gentian-violet being 

 superseded by methyl- violet 6.B. or methyl-violet B.N., and the Gram's fluid by that of Unna, 

 which yields nascent iodine, is given by Loeffler.-'* It is said to yield excellent results even 

 in unpractised hands. 



1 Plimmer, G., " Demonstrating Trypanosomata." Proceedings of Royal Society, Ser. B. LXXIX., pp. 95-102. 



^ Huisman, A. (1906) , "The Staining of Blood Films." MM. et Uijtj., No. 4. Quoted in British Mcdicnl Journal 

 Epitome, July 21st, 1906, p. 12. 



" Urtubey, A. (February 28th, 1907), " Note sur un precede simple, rapide et siir pour preparer le colorant de 

 Romanowsky-Leishman." Quoted in Btill. dc i'lnstitut Pnslciir, Vol. V. 



* Loeffler, F. (August 2nd, 1906), "Zur Qramschen F.irbungsmethode." Deiit. Med. Jfoch. Quoted in 

 Bull, de Vrmlilut Pasteur, October 30th, 1906, Vol. IV. 



• Article not consulted in the original. 



