Bacteria Staining 317 



Capsule Staining 



Hiss' Method (burrows, 1954) 



fixation: any good general fixative, 10% formalin is satisfactory. 



solutions: 



Either of 2 following stain solutions may be used: 



A. basic fuchsin, C.l. 42500 0.15-0.3 gm. 



distilled water 100.0 ml. 



B. crystal violet, C.I. 42555 0.05-0.1 gm. 



distilled water 100.0 ml. 



Copper sulfate solution: 



copper sulfate crystals 20.0 gm. 



distilled water 100.0 ml. 



procedure: 



1 . Deparaffinize and hydrate slides to water. 



2. Flood with either staining solution and heat gently until the stain 

 steams. 



3. Wash off the stain with copper sulfate. 



4. Blot, but do not dry. 



5. Dehydrate in 99% isopropyl alcohol, 2 changes: 1 minute each. 



6. Clear and mount. 



results: 



capsules — light pink (basic fuchsin) or blue (crystal violet) 

 bacterial cells — dark purple surrounded by the capside color 



Spirochete Staining 



Dieterle's Method (modified by beamer and firminger, 1955) 

 fixation: 10% formalin. 



solutions: 



Dilute gum mastic: 



gum mastic, saturated in absolute alcohol 30 drops 



95% ethyl alcohol 40.0 ml. 



Developing soliuion: 



distilled water 20.0 ml. 



hydroquinone 0.5 gm. 



sodium sulfite 0.06 gm. 



