We used Harleco's Wright-Giemsa, for human smears, it seems to work very
nicely for smears containing a lot of fat.  It was used by flooding the
slide with the Wright-Giemsa on a flat staining rack for 3 min, add water
or buffer, I would squirt water onto the slide with stain, using a squirt
bottle, I start at one end of the slide and proceed to the other end.

Watch it boil as it mixed, and even blew on the slide to make sure the mix
was well done, plus a green sheen should cover the whole top of this
mixture, let stand for 6 minutes, rinse with water and wipe the back of the
slide. Being an old Med Tech, I still preferred to let the hematology dept
do this staining, I hid out in my histo lab!     

When adding the buffer or water, a few drops of stain may come off slide,
don't panic, it still works.  If you need to use buffer, it is a phosphate
buffer at pH 6.8.  The med techs in the hematology dept liked this stain
better than a Wright stain.  Fixation occurs because of the methanol in the
staining solution, 

Check your pH of buffer or water, it may be off a tidge, and create
different tintorial quality, and if it is uneven, the mixing may be incomplete
Do you work with your hematology dept? They are very good at doing smears,
and were the ones who did all ours in the hospital, using the Harleco W/G.
I looked over their shoulders and observed. 

There are other Giemsa mixtures, also W/G on the market, and they seemed to
stain with some differences, why??  I couldn't tell you, maybe the original
sources of their dyes.  It was frustrating. 

A word of caution, make sure your staining solution is not old, we store
ours in the refrig (even though it says RT storage) we had an instance
where the solution was too old, and all went bust, found out it was past
expiration and had been used a lot (large bottle, open and closed many
times, decided the methanol content may have changed???? due to age and
evaporation, a new bottle saved the day)

Whew what an early Monday morning lecture!

 

Rande may be able to address this.
>Date: Mon, 06 Mar 2000 09:03:02 -0500
>From: rkline@emscience.com
>Subject: Re: bone marrow smears
>To: lizellis <lizellis@gateway.net>
>CC: Histonet <HistoNet@pathology.swmed.edu>
>
>Liz,
>
>I would like to help you.  EM Science has a couple of stains you can try.
I can
>send you a procedure and sample of Harleco's Wright-Giemsa or a Geimsa
Stain we
>import from Merck KGaA in Germany.
>You can call me at 800-222-0342x443 or e-mail me.
>
>Regards,
>
>Rande Kline, HT (ASCP)
>Technical Services
>EM Science?BDH
>
>
>
>
>lizellis <lizellis@gateway.net> on 03/05/2000 01:38:10 PM
>
>To:   Histonet <HistoNet@pathology.swmed.edu>
>cc:    (bcc: Rande Kline/EMI/Merck)
>Subject:  bone marrow smears
>
>
>
>Has anyone out there found a good manual staining method for bone marrow
>smears which accompany a bone marrow biopsy?
>We have tried Diff-Quik, a long Giemsa procedure and a Wright-Giemsa one
>step method, all with mixed reviews.
> The Pathologist can only seem to read the smear 50% of the time. We have
>even tried  the Hem-Tek automated stainer in our Hematology Dept., again
>with mixed reviews. Any insight would be greatly appreciated.
>Liz Ellis,
>Histology Manager
>Quest Diagnostics, Horsham ,PA
>
>
>
>
>
>
>
>
>
>
Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303