SECTION TWO 



C. Mallory's Aniline Blue - Orange G. 



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



Orange G . . . . . . . . 2 gm. 



Phosphomolybdic acid 1%. . . . 100 ml. 



D. Orange G o-i% in absolute alcohol. 



Technique : 



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



2. Sections, about 8/Lt 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. 



10. Rinse quickly with absolute alcohol. 



1 1 . Clear in xylol and mount. 



Results: 



Elastic fibres : red. Collagen : blue. Muscle fibres : pale orange 

 to dirty yellow. Cytoplasm : varying shades of yellow. Erythro- 

 cytes : golden yellow. Kerantinized material : bright yellow. 



Reference: Margolena, L. A. & Dolnick, E. H. (195 1). 



ORCEIN - ANILINE SAFRANIN 

 For elastic and connective tissue fibres 



Solutions required: 



A. Orcein (Unna) h to 1% in 80% 



alcohol . . . . . . . . 100 ml. 



Hydrochloric acid, cone. . . i ml. 



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