252 Cytoplasmic Elements (chap. 18) 



3. Rinse in 3 changes of distilled Avater. 



4. Counterstain in safranin, or like. 



5. Dehydrate, clear, and mount. 



results: 



enterochromaffin Sfranules — dark blue 

 nuclei — red 



comments: 



Lillie and his co-workers have been devoting several years to research 

 on the enterochromaffin granules along with melanins, etc. A fe^v of 

 these references are: Glenner and Lillie (1957); Lillie (1955, 1956A, 

 B, and C, 1957A, B, and C, 1960, 1961); Lillie et al. (1953); Lillie et 

 al. (1957); Lillie et al. (1961). 



Azo-Coupling Method (gurr, 1958) 



fixation: formalin or Bouin's. 



solution: 



Garnet reagent: 



Garnet GBC salt^ 0.5 gm. 



distilled water 100.0 ml. 



borax, saturated aqueous 2.5 ml. 



If it is necessary to purify the garnet GBC, see page 346. 



procedure: 



1. Deparaffinize and hydrate to water. 



2. Transfer to garnet solution: 30-60 seconds. 



3. Wash in running water: 30 seconds. 



4. Stain in Mayer's (or other) hematoxylin: 3 minutes, 



5. Wash in running water: 5 minutes. 



6. Dehydrate, clear and mount. 



results: 



aro^entaffin granules — red 

 nuclei — blue 

 background — light yello^v 



^ Imperial Chemical Industries, Manchester, England, is best source. Other sources: Dajac 

 Laboratories or American Cyanamid Company. 



