SECTION TWO 



Note: If it is desired only to stain the elastic fibres, omit No. 5, 

 and treat sections with acid alcohol for a few seconds before 

 washing with distilled water. 



ORCEIN - ANILINE BLUE - ORANGE G 

 A differential stain for elastic fibres, collagen, keratin, etc. 



I gm. 

 100 ml. 

 0-6 ml. 



49 ml. 

 0*5 ml. 



0-5 gm. 

 2gm. 

 100 ml. 



Solutions required: 



A. Orcein 



Alcohol 70% 



Hydrochloric acid, cone. . . 



B. Alcohol 50% 

 Hydrochloric acid, cone. . . 



C. Mallory's Aniline Blue - Orange G. 

 Aniline blue, aqueous 

 Orange G 

 Phosphomolybdic acid 1%.. 



D. Orange G 1% in absolute alcohol. 



Technique: 



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



2. Sections, about 8/x in thickness, are fixed to slides, dewaxed 

 with xylol and passed through the usual descending grades of 

 alcohol to distilled water. 



3. Stain for one and a half hours in the orcein solution in a 

 closed staining jar. 



4. Differentiate with solution B, controlling under the micro- 

 scope, until most of the pink is extracted from the sections. 



Note : The duration of the differentiation will vary according to 

 the nature of the material and to the thickness of the sections. 



5. Wash thoroughly with running tap water. 



6. Wash with distilled water. 



7. Immerse in solution C diluted with an equal volume of 

 distilled water, for one to two minutes. 



8. Rinse with 95% alcohol. 



9. Rinse with two lots of solution D. 



175 



