314 Microorganisms (chap. 20) 



Kinyoun's Carbol-Fuchsin Method (marti and 

 Johnson's modification, 1951) 



fixation: any general fixative, 10% formalin preferred. 



solutions: 



Carbol-fuchsin: 



basic fuchsin, CI. 42500 4.0 gm. 



phenol (carbolic acid) crystals 8.0 gm. 



95% ethyl alcohol 20.0 ml. 



distilled water 100.0 ml. 



Add 1 drop Tergitol :^7 to every 30 ml. of above. 

 Acid alcohol: 



nitric acid, concentrated 0.5 ml. 



95% ethyl alcohol 95.0 ml. 



Malachite green: 



malachite green oxalate, C.I. 42000 I.O gm. 



distilled water 100.0 ml. 



procedure: 



1. Deparaffinize and hydrate slides to water. 



2. Stain in carbol-fuchsin: 30 minutes, room temperature. 



3. Wash in running water: 5 mintites. 



4. Decolorize in acid alcohol: 3 minutes. Mixture should not be yel- 

 low. 



5. Wash in running water: 5 minutes. 



6. Rinse in 95% alcohol: 1 minute. 



7. Wash in running water: 2 minutes. 



8. Stain in malachite green: 0.5 minute. 



9. Wash in rtinning water: few minutes. 



10. Rinse in 95% alcohol, 10 dips. 



11. Dehydrate, absolute alcohol; clear and mount. 



results: 



acid-fast bacteria — red 



tisstie elements — shades of green 



comments: 



Instead of malachite green, LilHe's (1944) methylene blue can be 

 used; 1% methylene blue in 0.5% aqueous acetic acid: 3 minutes. 



Fite-Formaldehyde Method (wade's modification, 1957) 



fixation: Zenker's preferred; removal of HgClo not necessary, it dis- 

 appears during processing. 



