1314 Journal of Applied Microscopy 



MEDICAL NOTES. 



Ehrlich's Triacid Mixture for Staininc; Blood. 



Orange G, sat. aq. sol., .... 30. 



Saurefuchsin, " " .... 20. 



Methyl Green, " " .... 33. 



Alcohol, absolute, ..... 25. 



Glycerin, ....... 50. 



Water, dist., 75. 



Unless the solutions are absolutely saturated before mixing, results will be 

 unsatisfactory. In making up the mixture, the orange G and acid fuchsin are 

 first thoroughly mixed; then, drop by drop, the methyl green is added, the 

 solution being well shaken after each addition. The other three elements are 

 then added, and the whole shaken thoroughly. 



When the stock solution is once prepared it should never be shaken, but the 

 desired amount drawn from the top by means of a pipette. The specimen to be 

 stained is prepared in the usual manner by heat, and over the cover-glass spread 

 is placed a drop of the stain, which is allowed to remain for two or three minutes 

 or longer, if desired, after which the specimen is washed in water. Care must 

 be taken in the application of heat during the staining process, for if too much 

 heat is applied the specimen becomes pale yellow and is indistinct under the 

 microscope ; while if not enough heat is applied the specimen is too dark. 



After being thoroughly washed in water, the specimens are dried with filter 

 paper and mounted in Canada balsam. If the mixture is properly made it will 

 keep for years. c. w. j. 



Method for Cultivating and Staining the Diphtheria Bacillus ( Weiner 

 Medical Wocheiischi-ift, No. 10, 1900). — Twist a small piece of absorbent cotton, 

 impregnated with glucose glycerinated agar-agar, around the end of a sterilized 

 glass rod. A supply of these rods thus prepared may be kept on hand, each in 

 a sterilized test tube. To make a culture the cotton is swabbed over the affected 

 material, and the rod returned immediately to the test tube. After being kept in 

 the incubator at a temperature of 36° to 37° for four or five hours, enough bac- 

 cilli will have developed to make a smear. This is stained with the following 

 methylen blue solution : 



Methylen blue, ..... 1 gm. 



Alcohol, 20 c.c. 



Water, dist., 420 gms. 



Acetic acid, ..... 50 gms. 



This solution must not remain on the specimen more than two or three sec- 

 onds, after which time the slide should be thoroughly washed with water. For 

 a counterstain the following is used : 



Vesuvin, ...... 2 gms. 



Water, 1000 gms. 



which is heated, and filtered while still warm. 



The vesuvin should act on the specimen for fifteen to twenty seconds, being 

 then washed off with water. 



In absence of true bacilli, the smear appears brown. If both true and pseudo 

 bacilli are present, a blue and brown color is visible. The true bacilli stain 

 brown with their polar ends blue, while the pseudo bacilli stain wholly brown. 



c. w. J. 



