STAINING, PRACTICAL AND THEORETICAL 



4. Allow to dry in the air; then fix in absolute alcohol for fifteen 

 minutes. 



5. Pour off any excess alcohol, and allow the last traces to 

 evaporate away in the air. 



6. Stain for fifteen to thirty minutes in a mixture consisting of 

 10 ml. distilled water buflFered to 7-2, and 0-5 ml. Giemsa stain. 



7. Rinse in running tap water for a few seconds holding the 

 slide with the film facing downwards. 



8. Blot dry, and examine. 



Results: 



The spirochaetes are stained reddish violet. Nuclei of the leuco- 

 cytes reddish purple, while the rest of the leucocytes and pus cells 

 appear similar to Leishman-stained preparation. Cytoplasm of 

 Plasmodia, blue; chromatin, red. Erythrocytes: yellowish pink. 



Reference: Giemsa, G. (1904). 



GIEMSA STAIN - MAY-GRUNWALD STAIN 

 For blood and parasites 



Solutions required: 



A. May-Grunwald stain. 



B. Giemsa stain. 



Technique: 



1. Unfixed air-dried films are stained for three minutes in May- 

 Grunwald stain; then an equal volume of distilled water is added 

 and mixed with the stain by rocking the slide. The diluted stain is 

 then allowed to act for one minute; then drained off, without 

 washing. 



2. Stain for ten to fifteen minutes in a mixture consisting of 10 

 drops of Giemsa stain in 10 ml. distilled water. 



3. Differentiate for about five seconds, with distilled water, 

 examining under the microscope to ensure that differentiation is 

 complete. 



4. Blot and dry in air; mount. 



220 



