64 MICROSCOPIC TECHNIQUES 



PROCEDURE 



1. Fix for a short period in alcohol or 10% formalin. 



2. Prepare paraffin sections and employ egg albumin to hold to 

 slides. 



3. After removal of paraffin and bringing down to water, subject 

 sections to the iodine reagent for 6-12 hr. 



4. Wash in distilled water and decolorize with the sodium hypo- 

 sulfite for 15-30 sec. 



5. Wash in distilled water and stain with alum carmine for 1-3 

 hr. 



6. Wash in distilled water, dehydrate in acetone, clear in xylol, 

 and mount in balsam. 



Result. Bile pigments appear emerald green. Localizations can- 

 not be considered reliable due to the diffusibility of the reactants and 

 the final color. 



Forsgren Test for Bile Acids 



SPECIAL REAGENTS 



3% Barium Chloride. 

 0.1 % Acid Fuchsin. 

 1 % Phosphomolybdic Acid. 



Aniline Blue-Orange G Stain. Dissolve 0.5 g. aniline blue, 2 g. 

 orange G, and 2 g. oxalic acid in 100 ml. distilled water. 



PROCEDURE 



1. Treat small pieces of tissue for 6-12 hr. with the barium 

 chloride soln. 



2. Fix in 10% formalin for 12-18 hr. 



3. Prepare paraffin sections. 



4. Stain sections for 1-3 min. in the acid fuchsin soln., and 

 wash in distilled water. 



5. Place sections in the phosphomolybdic acid soln. for 0.5-1.0 

 min. and wash in distilled water. 



6. Treat sections for 3-5 min. with the aniline blue-orange G 

 staip and wash in distilled water. 



7. Dehydrate, clear, and mount in balsam. 



Result. Bile secretory granules appear reddish. 



