268 Cytoplasmic Elements (chap. 18) 



Working solution: 



carmine stock solution 30.0 ml. 



ammonium hydroxide 25.0 ml. 



methyl alcohol 25.0 ml. 



Lasts 2-3 weeks. 



Differentiating fluid. 



absolute ethyl alcohol 16.0 ml. 



methyl alcohol 8.0 ml. 



distilled water 20.0 ml. 



procedure: 



1. Deparaffinize and transfer into absolute alcohol: 3 mintites. 



2. Coat with 1% celloidin; dry slightly in air. 



3. Dip 2 or 3 times in 70% alcohol and then into water. 



4. Stain in Mayer's hematoxylin: 5 minutes. 



5. Wash, blue in Scott's, and wash. 



6. Stain, Best's carmine working soliuion: 15-30 minutes. 



7. Treat with differentiating fluid: 5-15 minutes. 



8. Rinse quickly in 80% alcohol. 



9. Dehydrate, clear, and mount. 



results: 



glycogen — red 

 nuclei — blue 



comments: 



Pearse (1953) proposes the following method of fixation, claiming 

 that it shows a localization of glycogen comparing favorably with that 

 preserved by freeze-drying. 



Lison's "Gendre fluid." 



picric acid, saturated in 96% alcohol 85 parts. 



formalin 10 parts. 



glacial acetic acid 5 parts. 



Cool to — 73 °C before use in an acetone-CO^. sno^v mixture. 



Use small pieces; fix for 18 hours. 



See also the Bauer-Feulgen reaction for glycogen, page 301. 



