Demonstration of tlio Bactorium in Tissue by a Modification of the Dieterle Silver Itiiprcgnation Stain 



REAGENTS 



1 . 5% alcoholic uranyl nitrate. Dissolve 50 gm uranyl nitrate in 1,000 ml of 70% alcohol. 

 Stable in refrigerator for months. 



2. 10% alcoholic gum mastic. Mix 100 gm gum mastic in 1,000 ml absolute alcohol, 

 and allow 2-3 days to dissolve. Filter solution and store in refrigerator in a well-stoppered 

 bottle. Gum mastic is available from O.G. Innes Corp., 10 East 40th Street, New York, 

 NY 10016. 



3. 1% silver nitrate. Dissolve 10 gm silver nitrate in 1,000 ml of distilled water. Store in 

 refrigerator. Discard solution if it turns dark. 



4. Developing solution - Mi.x in order: 



Hydroquinone 15.0 gm 



Sodium sulfite 2.5 gm 



Distilled water 600.0 ml 



Acetone 100.0 ml 



Formalin (3 7%-40%) 1 00.0 ml 



Pyridine 100.0 ml 



1 0% alcoholic gum mastic 1 00.0 ml 



Swirl flask gently as each solution is added. Solution turns milky yellow as the gum mastic 

 is added and light brown after standing in a well-lighted area. As brown streaks appear, 

 swirl the flask gently. Developing solution is ready to use in about 6 hours and can be used 



until it turns dark brown (usually in 2-3 days). 



REFERENCES 



1. Dieterle, R.R. 1927. Method for demonstration of Spirochaeta pallida in single microscopic sections. Archives 

 of Neurology and Psychiatry. 18:73-80. 



2. Van Orden, A.E., and P.W. Greer. 1977. Modification of the Dieterle spirochete stain. J. of Histotechnology. 

 1:51-53, 



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