SECTION TWO 



Aniline Blue - Orange G (Mallory): 

 D. Aniline blue, aqueous . . . . 0-5 gm. 



Orange G. . . 

 Distilled water 



E. Solution D . . 

 Distilled water 



. . 2 gm. 

 100 ml. 



I volume 

 2 to 3 volumes 



Technique: 



1. Fix thin slices of pancreas in Bouin for eight to ten hours. 



2. Wash in distilled water, dehydrate, clear and embed in 

 paraffin wax. 



3. Cut section 4/x in thickness. 



4. Fix section to slides; dewax and take down through the 

 usual grades of alcohol to distilled water. 



5. Stain in solution A for about forty-five to sixty minutes at 



56° c. 



6. Rinse quickly in distilled water and blot very carefully. 



7. Destain in solution B until acinous tissue is almost colourless 

 and B cells show red against pink background of A cells. 



8. Rinse briefly with distilled water and treat with 5% iron 

 alum solution for 5 minutes or more. 



9. Rinse again and stain two to twenty minutes in solution E 

 until the collagenic tissue appears deep blue under the microscope. 



10. Rinse and blot carefully. 



11. Differentiate and dehydrate in absolute alcohol. 



12. Clear in xylol and mount. 



Results : 



Cytoplasm of A cells: rich yellow orange; of B cells: bright 

 red ; and of D cells : sky blue. 



Note : It is stated that it can be demonstrated that there is no gradation 

 between A and B cells by first staining with Neutral Gentian (Bensley) 

 decolorizing, then restaining by the above technique. 



Reference : 



Gomori, G. (1939), Anat. Rec, 74, 439-459. 

 Cowdry, E. V.: Laboratory Technique, 3rd cd., p 167. 



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