Trichrome Staining 157 



Acidic water: 



glacial acetic acid 0.2 ml. 



distilled water 100.0 ml. 



procedure: 



1. Deparaffinize and hydrate slides to water; remove HgCL. 



2. Slightly overstain with any hematoxylin. 



3. Wash in running water: 5 minutes. 



4. Stain in trichrome mixttire: 5-20 minutes. 



5. Rinse briefly in distilled water to remove some of excess stain. 



6. Rinse briefly in acidic water. 



7. Rinse in 70% alcohol. 



8. Dehydrate in 95% alcohol, 2 changes; be careful not to remove 

 too much green. 



9. Dehydrate, absolute alcohol, clear and mount. 



RESULTS AND COMMENT 



There have been a number of attempts to combine the azan and Mas- 

 son methods in a quicker one. Gomori considered Pollak's as a disap- 

 pointingly dull color scheme, lacking real red shades. Almost any 

 combination of an acid triphenylmethane dye with a sulfonated azo 

 dye in the presence of phosphomolybdic or phosphotungstic acid will 

 give good results. The former favors the green and blue shades, the 

 latter the reds. A short staining time produces more red and pro- 

 longed staining more green and blue. Rinsing in tap water weakens 

 the reds. 



If more red is desired — in muscle and epithelial fibers, for ex- 

 ample — before staining treat with warm Bouin's 56-60°C: 2-5 min- 

 utes. Follow by washing until yellow color disappears. 



The rinse in acetic acid does not change the colors, btit makes them 

 more delicate and transparent. 



Lillie {1954B) says that Gomori's trichrome gives a rather diffuse 

 and incomplete picture of the more finely fibrillar stroma, and the 

 reticulum of the liver and spleen are difficult to discern. 



The author has had good results with most of the trichrome meth- 

 ods and can only recommend that the selection of a routine stain will 

 have to be made by each individual. All of the short trichrome meth- 

 ods offer clearer and sharper returns on thin (5-7 microns) sections 

 than on thicker ones. For the latter, Mallory's, Masson, or Azan meth- 

 ods shotdd be used. 



