328 Microorganisms (chap. 20) 



procedure: 



1. Hydrate slides to water (these will be smears). 



2. Stain in working solution, 37 °C: 1 hour. 



3. Rinse rapidly, 2 changes 95% ethyl alcohol. 



4. Dehydrate in absolute alcohol, clear, and mount. 



results: 



inclusion bodies — blue to purplish blue 



Negri Bodies (Rabies) 



Schleif stein (1937) Method 



fixation: Zenker's preferred. 



solutions: 



Staining solution: 



basic fuchsin, C.I. 42500 1.8 gm. 



methylene blue, C.I. 52015 1.0 gm. 



glycerol 100.0 ml. 



methyl alcohol 100.0 ml. 



Keeps indefinitely. 



For use add about 10 drops to 15-20 ml. of dilute potassium hydrox- 

 ide (1 gm./40,000 ml. water). Alkaline tap water may be used. 



procedure: 



1 . Deparaffinize and hydrate slides to water; remove HgCl2. 



2. Rinse in distilled water and place slides on warm electric hot plate. 



3. Flood amply with stain and steam for 5 minutes. Do not allow 

 stain to boil. 



4. Cool and rinse in tap water. 



5. Decolorize and differentiate each slide by agitating in 90% ethyl 

 alcohol until the sections assume a pale violet color. This is im- 

 portant. 



6. Dehydrate, clear, and mount. 



results: 



Negri bodies — deep magenta red 

 granular inclusions — dark blue 

 nucleoli — bluish black 

 cytoplasm — blue-violet 

 erythrocytes — copper 



comments: 



Schleifstein outlines a rapid method for fixing and embedding so the 



