Rickettsia and Virus Staining 327 



results: 



rickettsiae, also inclusion bodies — bright red 

 nuclei and cytoplasm — sky blue 



Giemsa Method (American public health association, 1956) 

 fixation: general fixative, Regaud recommended. 



solutions: 



Giemsa stock solution, see page 225. 



Buffer solutions: 



A. dibasic sodium phosphate, anhydrous, Nao 



HPO4 9.5 gm. 



distilled water 1000.0 ml. 



B. monobasic sodium phosphate, NaH2P04- 



H.O 9.2 gm. 



distilled water 1000.0 ml. 



Working solution, pH 7.2: 



solution A 72.0 ml. 



solution B 28.0 ml. 



distilled water 900.0 ml. 



Giemsa working solution: 



Dilute 1 drop Giemsa stock solution with 5 ml. of buffer working 

 solution. 



procedure: 



1. Deparaffinize and hydrate slides to water. (Smears can be carried 

 directly to water: wash well in running water: 5-10 minutes.) 



2. Leave slides in Giemsa working solution overnight, 37 °C. 



3. Rinse thoroughly in distilled water. Dry between blotters. 



4. Dip rapidly in absolute alcohol. If overstained, use 95% alcohol 

 to decolorize them, dip in absolute alcohol. 



5. Clear and motmt. (Smears, after treatment with absolute alcohol, 

 can be washed in distilled water: 1-2 seconds, and blotted dry. 

 Examine with oil or mount with a cover glass.) 



results: 



rickettsiae and inclusion bodies (psittacosis) — blue to purplish blue 



Modification for trachoma. 



SOLUTION : 



Giemsa working solution: 



Dilute 1 drop of stock Giemsa with 2 ml. of buffer solution above. 



