SECTION TWO 



BIEBRICH SCARLET - ETHYL VIOLET - HAEMATOXYLIN 



(Cambel and Sgouris modification of Bowie's Stain) for 

 pepsinogen granules of the body chief cells in the gastric 



glands 



Solutions required: 



A. Delafield Haematoxylin, aqueous 



B. Ethyl Violet - Biebrich Scarlet 

 20 % Ethyl alcohol . . 



C. Solution B. . . 

 Absolute alcohol 



Distilled water . . 



0.5 gm. 

 50 ml. 



0-5 gm. 

 20 ml. 



80 ml. 



N.B. — This solution must be freshly pre- 

 pared each time it is required for use. 



D. Clove Oil . . . . . . . . I volume 



Toluol . . . . . . . . I volume 



Technique: 



1. Fix in Regaud's fluid for five days in a dark phial placed in a 

 larger amber bottle which should be wrapped round with a thick 

 cloth and kept in a dark room. Change the Fixative daily. 



2. Dehydrate with normal propyl alcohol (see page 38). 



3. Transfer to paraffin wax which should be changed three times 

 before the block is finally cast. 



4. Cut sections 5 to 6/x in thickness and fix to slides as usual. 



5. Remove wax with two changes of xylol. 



6. Pass through absolute alcohol and the usual ascending grades 

 of alcohols down to distilled water. 



7. Stain in the Haematoxylin solution for i minute. 



8. Wash and blue in tap water. 



9. Remove excess water by draining and blotting very carefully. 



10. Stain with solution B for ten to fifteen minutes, or longer 

 (up to twenty-four hours). 



11. Rinse briefly in distilled water, then drain and carefully 

 remove excess water by blotting. 



G yi 



