Proceedings of 

 the United States 

 National Museum 



SMITHSONIAN INSTITUTION • WASHINGTON, D.C. 



Volume 122 1907 Number 3596 



An Enzyme Method 



Of Clearing and Staining 



Small Vertebrates 



By William Ralph Taylor 

 Associate Curator, Division of Fishes 



Introduction 



Cleared (transparent) and stained specimens are used widely for 

 osteological studies of vertebrates. Many such specimens have been 

 prepared by the Schultze method (Schultze, 1897) or modifications of 

 that method (Davis and Gore, 1947; Evans, 1948; Hollister, 1934; 

 Jensh and Brent, 1966). The Schultze method and its modifications 

 chiefly involve tissue maceration in solutions of potassium hydroxide 

 or sodium hydroxide, staining the bones to give them maxunum visi- 

 bility, and immersion of the specmien in a liquid with an index of 

 refraction similar to that of the cleared tissues. 



The method is successful when it is used on fresh or newly preserved 

 specimens, but numerous workers have encountered considerable 

 difficulty in producing good transparent specimens by the above 

 methods after specimens have been stored m various preservatives 

 for several years. Such specimens may remain opaque because of dark 

 stains in the flesh or they may swell and the membranes rupture dur- 

 ing the clearing process. 



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