Bacteria StaiJiijin ^10 



'rt 



No contamination; coat forceps with paraffin. 



Solutions must be fresh (no more than 1 week old) and made from 

 triple distilled water. 



Carry a kno^vn positive control slide with test slide through the 

 process, or preferably a control section on the same slide. 



solutions: 



Acidulated water: 



triple distilled water 1000.0 ml. 



citric acid 10.0 gm. 



pH 3.8-4.4 



2% Silver nitrate (for developer): 



silver nitrate 2.0 gm. 



acidulated water 100.0 ml. 



1% Silver nitrate: 



dilute a portion of above with equal volume of acidulated water. 



0.15% Hydroquinone: 



hydroquinone 0.15 gm. 



acidulated water 100.0 ml, 



5% gelatine: 



sheet gelatine or granulated of high degree of 



purity 10.0 gm. 



acidulated water 200.0 ml. 



Developer: 



2% silver nitrate 1 .5 ml. 



5% gelatine 3-75 ml. 



0.15% hydroquinone 2.0 ml. 



Warm solutions to 55-60°C and mix in order given, with stirring. 

 Use immediately. 



procedure: 



1 . Deparaffinize and hydrate slides to acidulated water. 



2. Impregnate in 1% silver nitrate, 55-60°C: 30 minutes. 



3. Place slides on glass rods, pour on warm developer (55-60°C). 

 When sections become golden bro^vn or yellow and developer 

 brownish-black (3-5 minutes) pour off. The known positive can be 

 checked under microscope for black organisms. 



4. Rinse with warm (55-60°C) tap w^ater, then distilled water. 



5. Dehydrate, clear, and mount. 

 results: 



spirochetes — black 



