MUCOPROTEINS, GLYCOGEN, AND MUCIN 47 



A fundamental study of metachromasy of basic dyes has been 

 published by Miehaelis and Granick (1945). 



See page 45 for another method of staining acid polysaccharides, 

 and page 50 for the staining of mucin. 



Lison Method for Polysaccharide Sulfate Compounds 

 (after Sylven) 



SPECIAL REAGENTS 



Fixing Solution. Mix equal vol. of 8% basic lead acetate soln. 



and 14-16% formalin. 

 Toluidene Blue Solution. Prepare separately (a) 0.1% dye in 1% 



alcohol and (6) 0.1% dye in 30% alcohol, and let stand for a 



number of days to age. 



PROCEDURE 



1. Fix the tissue for 12-24 hr. in the fixing soln. 



2. Prepare paraffin sections in the usual manner. 



3. Stain the sections for 30 min. using soln. a on some, and soln. 

 b on others. Soln. a gives a more intense stain. 



4. Wash well in alcohol briefly, immediately after removing from 

 the dye soln. 



5. Mount in natural cedar oil. 



GLYCOGEN AND MUCIN* 



A critical comparison of the iodine, Best carmine, and Bauer- 

 Feulgen methods for demonstrating glycogen microscopically was 

 ,made by Bensley (1939), who concluded that the Bauer-Feulgen 

 method, which depends on the reaction of the aldehyde groups in 

 the carbohydrate with the reagent, is by far the best if the tissue 

 is promptly fixed in alcohol-formalin solution. When chrome salts 

 are present in the fixative, the visualization of intracellular glycogen 

 was found to require the Best carmine stain since the Bauer-Feulgen 

 method is not specific in those cases, and the iodine technique is 

 not suited for high-power studies. A procedure for preparing paraffin 

 sections for the carmine stain was given by Mullen (1944), who 

 employed celloidin to hold the deparaffinized sections to the glass 

 slide. 

 Mitchell and Wislocki ( 1944) reported that the ammoniacal silver 



* See Bibliography Appendix, Refs. 2 and 12. 



