Re: May Grunwald - Giemsa problems!

You should fix the slides before you are ready to stain them.  Make sure
the methanol is fresh.  If the bottle has been open for a long time most
likes water has gotten into it.  Use a new bottle is this is true.
Argentina is probably fairly humid.  In which case you should also change
the methanol frequently.  I'm not sure why you are getting green RBC's.
That's a new one for me.
What is your procedure and who's stains are you using?

marina goumenou  on 10/03/2002 04:30:29 PM

Subject:    May Grunwald - Giemsa problems!

Hi to all Histonetters

I work with May Grunwald - Giemsa for about 2 years staining bone marrow
smears from rats (I count micronucleus in polychromatic RBC) but the
are not constantly well. So, I still search to find what is going whrong.
Some of the artifacts I have noticed are:
Very bright RBC, green RBC, no diferentiation between normochromatic and
polychromatic RBC, breaked RBC, purple in the center of normochromatic RBC
etc. I thought many different possible reasons for all these problems and I
try to stabilize the parameters of the protocol I use but I am not sure
about my thoughts:
I use DI water instead of buffer, is this sufficient? (I am not sure that
water is constantly well)
I filter Giemsa before use but what kind of filter is appropriate (pore
size, material etc)?
I use stock solutions (botlles of 2.5L) and some times they are not in use
for weeks after I open them. Is this a possible problem?
How long can I store my smears after fixation (5min in methanol) without
staining them?
Is it possible having burst RBC due to FBS osmolarity and/or water in
methanol during fixation?

I really need any help, any thought, any technical information!
Thanking you in advance
Goumenou Marina

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