Triclirome Slainitw l*'l 



i--) 



Kornhauser's Quad Stain (1043, 1945) 



fixation: any general fixative, preferably containing HgClo. 



solutions: 

 Orcein: 



orcein 0.4 gm. 



nitric acid, cone 0.4 ml. 



90% ethyl alcohol 100.0 ml. 



Acid alizarine blue: 



acid alizarine blue BB, C.I. 58610 0.35 gm. 



aluminum ammonium sidfate 5.0 gm. 



acetic acid, N/10 (5.8 ml. glacial acetic acid 



made up to 1 liter) 82.0 ml. 



Boil gently in Erlenmeyer flask covered with watch crystal: 10 min- 

 utes. Cool and filter. 



Phosphotinigstic-phosphoniolybdic acid: 



phosphotungstic acid 4.0 gm. 



phosphomolybdic acid 1.0 gm. 



distilled water 100.0 ml. 



Orange G-fast green: 



orange G, C.I. 16230 2.0 gm. 



fast green FCF, C.I. 42053 0.2 gm. 



glacial acetic acid 2.0 ml. 



distilled water 100.0 ml. 



procedure: 



1. Deparaffinize and run slides down to 85% alcohol. Remove HgCl. 

 if present. 



2. Orcein: 2-24 hours, until elastin fibers are red brown. May be 

 oinitted if no elastin is present or is not to be stained. 



3. Wash otit excess orcein, two changes 85% alcohol. 



4. Rtui down to water. 



5. Acid alizarine blue: 5-10 minutes. 



6. Decolorize and mordant in phosphotungstic-molybdic sohition, 

 tintil collagen is destained: 10-30 minutes. 



7. Rinse few seconds in distilled water. 



8. Stain in orange G-fast oreen: 3-10 minutes. 



9. Rinse in 50% alcohol: about 10 seconds. 



10. Dehydrate, 95% and absoliue alcohol, clear aird mount. 



