1 8 MEDICAL BACTERIOLOGY 



ANILIN WATER 



Anilin water is prepared by adding anilin oil to distilled water, shaking after 

 the addition of each drop. The oil is added until the water is saturated. This 

 water must be filtered until entirely free from visible globules of oil, even though 

 several passages through filter-paper be necessary. 



ANILIN GENTIAN VIOLET 



Anilin water, 84 c.c. 



Saturated alcoholic solution of gentian violet, 16 cc. 



Anilin gentian violet rapidly deteriorates and in a few days or weeks is 

 useless. 



GRAM'S SOLUTION 



Iodine, i Gm. 



Potassium iodide, 2 Gm. 



Distilled water, 3 oo cc. 



In routine work Loeffler's methylene blue is generally used and found most 

 satisfactory for staining the diphtheria bacillus, but one of the characteristics 

 of this organism, irregular staining, is best brought out by Neisser's method 

 of staining. 



When stained with Loeffler's methylene blue, one, two or more portions 

 of the diphtheria bacillus stain de eply, the rest of the organism stains faintly or 

 not at all, thus giving a deceptive impression of its contour. With the two 

 extremities stained deeply and the intermediate portion stained faintly or not 

 at all, so-called polar staining, or with both poles and the center stained deeply 

 and the portions between the center and poles unstained, so-called barred stain- 

 ing or beaded forms, the organism may appear more like several cocci in a row 

 than a bacillus, which it really is. 



By Neisser's method the entire bacillus is stained, the ends or the ends and 

 center blue, intermediate portions light brown. Two solutions are used to 

 obtain this effect, as follows: 



NEISSER'S METHOD 



1. Apply Neisser's acid methylene blue to specimen for 2 to 5 minutes. 



2. Wash with water to remove excess stain. 



3. Apply Bismarck brown for i minute. 



4. Wash with water. 



5. Dry, mount and examine. 



NEISSER'S ACID METHYLENE BLUE 



1. Methylene blue, o.i Gm. 3. Glacial acetic acid, 5 cc. 



2. Alcohol, 2 cc. 4. Distilled water, 95 cc. 



. First mix the methylene blue and alcohol together, then mix the glacial 

 acetic acid and distilled water together, finally add the acid solution to the 

 alcoholic solution, and filter. 



