Elizabeth M. Cameron, HT(ASCP), QIHCCM. Lead Histologist at Mid Coast Hospital in Brunswick, Maine asks:
>>I have been staining fish tissues fixed in Davidson's fixative with H&E, and the researcher would like the eosin to be more intense. Our standard protocol works well for our own tissue, but the fish look much more washed out. I am using alcoholic eosin Y, have tried both water and alcohol before and I have varied the alcohol differentiation steps after the Eosin. I also extended the time in Eosin and increased the wash after bluing to make sure the sections are not basic. Any suggestions would be appreciated.<< Users vary a lot in how much eosin they want. Sometimes blends with other red dyes are preferable. When I was a resident at Johns Hopkins around 1970 our notoriously lurid eosin was compounded as follows: Eosin Y (C.I. 45380) 3.6 g Phloxine B (C.I. 45410) 1.5 g Biebrich scarlet (C.I. 26905 ) 0.3 g absolute alcohol 150 mL distilled water 120 mL Shake, or stir with a magnetic stirrer to dissolve, then add 450 mL more of distilled water. We used a lot of Davidson's fixative at JHH. As far as I know this formula was never published, though I've posted it online more than once, probably on Histonet. One of many formulas I made off with before I finished residency. Bob Richmond Samurai Pathologist Maryville TN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet