GIEMSA'S STAIN 213 



3. Add water to the stain on the cover-glass or slide, drop by drop, until a yellow 

 metallic scum begins to form. It is advisable to add the drops of water rapidly in 

 order to eliminate precipitates on the stained film. Practically, we may add i drop of 

 water for every drop of stain used. 



4. Wash thoroughly in water until the film has a pinkish tint. 



5. Dry with filter-paper and mount. The stained preparation is less apt to show 

 foreign material and damage if one allows the film to dry without blotting. In a 

 moist atmosphere one may dry the film high over the flame but any near contact 

 with the heat of the flame is injurious. 



Red cells are stained orange to pink; nuclei, shades of violet; eosino- 

 phile granules, red; neutrophile granules, yellow to lilac; blood platelets, 

 purplish; malarial parasites, blue; chromatin, metallic red to rose pink. 



The bottle in which the methyl alcohol solution is kept must be tightly stoppered 

 with a cork stopper and kept in the dark. Any evaporation of the alcohol interferes 

 with proper fixation of the blood film and the light affects the delicate stain. 



Giemsa's Modification of the Romanowsky Method. This is one of 

 the most perfect of the modifications. The objection is that greater 

 time in staining films is required than with the Wright or Leishman 

 method and the stain is very expensive. 



Take of Azur II eosin 0.3 gram. Azur II 0.08 gram. 



Dissolve this amount of dry powder in 25 c.c. of pure anhydrous glycerine at 6oC. 

 Then add 25 c.c. of methyl-alcohol at the same temperature. Allow the glycerine 

 methyl-alcohol solution to stand over night and then filter. This is the stock stain. 

 To use: Dilute i c.c. with 10 to 15 c.c. of distilled water. If i to 1000 potassium 

 carbonate solution is used instead of water it stains more deeply. 



The alkaline diluent is used to obtain the course stippling in malig- 

 nant tertian (Maurer's clefts). Having fixed the smear with methyl 

 alcohol for one to five minutes, pour on the diluted stain, and after 

 fifteen to thirty minutes wash off and continue washing with distilled 

 water until the film has a slight pink tinge. For Treponema pallidum 

 stain from two to twelve hours. 



While the Romanowsky methods are more satisfactory for differential counts and 

 for the demonstration of the malarial parasites, and especially for differentiating 

 species, yet by reason of the liability to deterioration in the tropics of methylene 

 blue the haematoxylin methods may be preferable. Many workers in blood-work 

 and cytodiagnosis prefer the haematoxylin. 



1. Fix the film either by heat, with methyl alcohol for two minutes or with Whit- 

 ney's fixative. Heat is to be preferred. 



2. Stain with Meyer's hemalum or Delafield's haematoxylin for from five to 

 fifteen minutes according to the stain. Frequently three minutes will be found 

 sufficient. To make the hemalum, dissolve 0.5 gram of haematin in 25 c.c. of 95% 



