Bdcieria Slaiyiing .^15 



solutions: 



Phenol new fuchsin: 



new fuchsin, C.I. 42520 (Magenta III) 0.5 gni. 



phenol (carboHc acid) 5.0 gm. 



ethyl or methyl alcohol 10.0 ml. 



distilled water to make 100.0 ml. 



Van Gieson, modified: 



acid fuchsin, C.I. 42685 0.01 gm. 



picric acid 0.1 gm. 



distilled water 100.0 ml. 



procedure: 



1. Deparaffinize in turpentine-paraffin oil (2:1) 2 changes: 5 min- 

 utes total. 



2. Drain, wipe off excess fluid, blot to opacity, place in water. 



3. Stain overnight in phenol-fuchsin, room temperature. 



4. Wash in tap water. 



5. Treat with formalin (full strength): 5 minutes (turns blue). 



6. Wash in running water: 3-5 minutes. 



7. Treat with sulfuric acid, 5% (5 ml./95 ml. water): 5 minutes. 



8. Wash in running water: 5-10 minutes. 



9. Treat with potassium permanganate, 1% (1 gm./lOO ml. water): 

 3 minutes. 



10. Wash in rimning Avater: 3 minutes. 



11. Treat with oxalic acid, 2-5% (2-5 gm./lOO ml. water) individ- 

 ually with agitation: not more than 30 seconds. Slides can remain 

 in water while others are being treated. 



12. Stain with modified Van Gieson: 3 minutes. No wash. 



13. Rinse for few seconds in 95% alcohol. 



14. Dehydrate, clear, and moimt. 



results: 



acid-fast bacteria — deep blue or blue black 



connective tissue — red 



other tissue elements — yellowish 



comments: 



Beanier and Firminger (1955) emphasize care in the use of formalin. 

 Old formalin yields poor results, while a redistilled form produces 

 the most brilliant staining. The reagent grade in 16 oz. brown bottles, 

 if kept tightly closed, give a good stain. 



Tilden and Tanaka (19^5) outline a method for frozen sections, 

 essentially the same as the one above after moimting the sections ^vith 



