ROMANOWSKY STAINING METHODS. 151 



results, the addition of a drop or so of either the acid or alkaline 

 alcohol, as indicated, will restore its original staining properties. The 

 acid and alkaline alcohols may be used in the same way and with 

 about the same results with Wright's staining fluid. 



In all Romanowsky methods distilled water should be used. If not 

 obtainable, the best substitute is rain-water collected in the open and 

 not from a roof. 



Method of staining: 



1. Make films and air dry. 



2. Cover dry film preparation with the methyl-alcohol stain for i 

 minute (to fix). 



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. 



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

 eosinophile granules, red; neutrophile granules, yellow to lilac; blood 

 platelets, purplish; malarial parasites, blue; chromatin, metallic red to 

 rose-pink. 



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 gm. Azur II 0.08 gm. 



Dissolve this amount of dry powder in 25 c.c. of glycerin at 60 C. 

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

 the glycerin 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 water. If i to 1000 potassium carbonate solution is used instead of 

 water it stains more deeply. Having fixed the smear with methyl 

 alcohol for 5 minutes, pour on the diluted stain and after 15 to 30 min. 

 wash off and continue washing with distilled water until the film has 

 a slight pink tinge. For Treponema pallidum stain from 2 to 12 hours. 



