I would think more than pH is not proper with the stain solutions that are 20
years old.  That's old!

But, lets work on the pH factor first.  You did not state you staining procedure
or what you are using for decolorizing the smears.  Are you using a buffer, DI
water, or tap water and at what pH ?
You may also be decolorizing the slides too long.

Rande Kline
EM Science




"Agust?n Venzano"  on 03/19/2002 08:09:28 AM

To:   Histonet English 
cc:    (bcc: Rande Kline/EMI/Merck)
Subject:  Blood smears staining



Dear Histonet pals: Argentine Veterinary Pathologists do love our work, but
we are lacking funds, so we often have to manage just with the things we
dispose of, no matter how old they are. So I regret to post this message,
but I have to: Our May Grunwald and Giemsa solutions were purchased some 20
years ago, so here's the problem. When you stain a blood smear you can find
all basophilic structures as expected, i.e. nucleuses, lymphocytes'
cytoplasms, etc. But there's a lot of problem with eosinophilic things, i.e.
eosinophils' granules appear as unstained refractory ones, neutrophils'
cytoplasms appear more basophilic than usual, and red cells seem less
eosinophilic than normal ones. The detail of all cells is well preserved. So
I'm thinking that the solutions' pH has changed. Do you agree with me or
not? How should I repair the fault?

Thanks a lot for your attention.

Agustin Venzano Halliburton
Laboratory of Veterinary Histopathology
INTA Castelar, Argentina