324 Microorganisms (chap. 20) 



solutions: 



Acridine orange: 



acridine orange 0.1 gm. 



distilled water 100.0 ml. 



Weigert's hematoxylin, see page 126. 



procedure: 



1. Deparaffinize and hydrate sections to water. 



2. Stain in Weigert's hematoxylin: 5 minutes. 



3. Wash in running water: 5 minutes. 



4. Stain in acridine orange: 2 minutes. 



5. Rinse in tap or distilled water: 30 seconds. 



6. Dehydrate in 95% alcohol: I minute. 



7. Dehydrate in absolute alcohol, 2 changes: 2-3 minutes. 



8. Clear in xylene, 2 changes: 2-3 minutes. 



9. Mount in a nonfluorescing medium. 



results: 



all fungi fluoresce except Nocardia and Rhizopus; colors of the fluo- 

 rescing genuses appear as follows: 

 Coccidioides, Rhinosporidum — red 

 Aspergillus — green 



Actinomyces, Histoplasma — red to yellow 



Candida, Blastomyces dermatitids, Monosporium — yellow-green 

 Blastomyces hrasiliensis — yellow 



comments: 



Old hematoxylin and eosin slides may be decolorized and restained 

 as above. 



The Weigert's hematoxylin staining is necessary as a quenching 

 agent, because some fungi are difficult to see against a background 

 which also fluoresces. With the hematoxylin, the fungi fluoresce 

 brightly against a dark setting. 



Rickettsia and Inclusion Bodies 



Modified Pappenheim Stain (castaneda, 1939) 



fixation: any general fixative, Regaud recommended. 



solutions: 



Stock Jenner stain: 



Jenner's stain 1.0 gm. 



methyl alcohol, absolute 400.0 ml. 



