[Histonet] Eosin

[Bob Richmond]

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