STAINING, PRACTICAL AND THEORETICAL 



Technique : 



1. Fix slices of material for twenty-four hours in formalin- 

 calcium. 



2. After washing, dehydrate, clear, and embed in paraffin wax. 



3. Cut sections at 6 to 7/x and fix them to slides. 



4. Remove wax with xylol and pass through the usual descending 

 grades of alcohol to distilled water. 



5. Stain in the Maxilon blue RL solution for thirty to sixty 

 seconds. 



6. Wash in distilled water. 



7. Remove the excess water from the slide with filter paper. 



8. Immerse in two lots of anhydrous tertiary butyl alcohol 

 for one minute in each 



or 



Rinse in 70% alcohol and dehydrate in two changes of absolute 

 alcohol, for two minutes in each, but the tertiary butyl alcohol 

 procedure is considered to be the more satisfactory one. 



9. Clear in xylol. 



10. Mount in D.P.X. or Clearmount or Canada balsam in 

 xylol. 



Results: 



Acid mucopolysaccharide-containing elements are stained red 

 to violet, while other basophilic structures are stained in various 

 shades of blue. 



Notes: 



(a) The author states that his preliminary unpublished results 

 point to the possibility of distinguishing between the various 

 acid mucopolysaccharides present in tissue sections by varying the 

 dye concentration. 



(b) Maxilon blue RL is a basic dye of the mono-azo group. 

 Its 0-05% aqueous solution, which is dark blue in colour, has a 

 pH of 4-1. 



(c) It is also stated that this is similar to the method suggested 

 by Hempelmann (1940) with toluidine blue, which has apparently 



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