SECTION TWO 



ALCIAN BLUE 

 For acid mucopolysaccharides 



Solutions required: 



A. Alcian blue i % aqueous 



B. Haemalum 



Technique: 



1. Fix material in Bouin and embed in paraffin wax. 



2. Fix sections to slides and carry through to distilled water as 

 usual. 



3. Stain for fifteen to forty-five minutes in the alcian blue 

 solution. 



4. Rinse in distilled water. 



5. Stain in the haemalum solution for five to ten minutes. 



6. Counterstain if desired. 



7. Dehydrate through the usual graded alcohols. 



8. Clear in xylol and mount in D.P.X. or Clearmount. 



Results: 



Acid mucopolysaccharides are stained blue-green, while nuclei 

 are dark blue. 



Notes: 



(a) Alcian blue stains mucin clearly and conspicuously, com- 

 bining with it in such a way that additional stains produce little, 

 if any, alteration in colour. 



(b) All common histological reagents will not remove alcian 

 blue from mucin: only prolonged treatment with acid alcohol 

 will reduce the colour of an overstained section. Once applied, 

 the dye will resist indefinitely water, alcohols, alkalis, and hydro- 

 carbons. 



(c) Alcian blue will not distinguish chondroitin sulphuric acid 

 complexes from mucoitin sulphuric acid: the dye, therefore, 

 stains cartilage and mucin equally. 



(d) A crystal or thymol or a few drops of chloroform should be 

 kept in the alcian blue solution to inhibit the growth of air-borne 

 micro-organisms: it is best to prepare the solution in small 

 quantities as required for a day's use, however. 



Reference: Steedman, H. E. (1950). 



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