Augustin brought to my attention he was referring to blood smears.  They
still can be destained in 0.05% acetic water or a low percentage alcohol.
They should decolorize very quickly (in a matter of seconds). They will
need to be washed in di water, dried, and placed in methanol again.

Augustin- I trouble-shoot Giemsa a alot.  By chance are you using stains
from Merck since you are in Argentenia.  If not., who's stains do you use
and what is your procedure.   Believe it or not, it almost sounds as if
they are not being differentiated enough by your buffer.  Are you using a
buffer.  What pH is your buffer?

Rande Kline
EM Science




"Marshall Terry Dr, Consultant Histopathologist"
 on 09/06/2002 10:56:35 AM

To:    Agustín Venzano , Histonet Server
       
cc:
Subject:    RE: May-Grünwald-Giemsa


Agustin,
First try to ensure that the smears are thin and dried instantaneously - no
slower. I always shake the slide in the air like fury, usually to the
amusement of onlookers, but it is so essential. The speed of drying affects
the staining enormously.

Terry L Marshall B.A.(Law), M.B.Ch.B., F.R.C.Path
Consultant Histopathologist
Rotherham General Hospital, Yorkshire
terry.marshall@rgh-tr.trent.nhs.uk

-----Original Message-----
From: Agustín Venzano [mailto:avenzano@cicv.inta.gov.ar]
Sent: 06 September 2002 15:17
To: Histonet Server
Subject: May-Grünwald-Giemsa


Histonetters: I've lately been having some problems with blood films
staining. Despite previously filtering all reagents and strictly following
the required steps the results were frustrating.

Leukocytes' nuclei often appeared faintly stained, and their cytoplasms'
aspect was variable. Sometimes they looked well colored, but often
cytoplasmic coloration resulted dull. Moreover, eosinophils didn't show
there red cytoplasmic granules, instead their cytoplasm appeared rather
basophilic.

Of course I decided to immediately dispose of MG and G solutions in use,
but
now I need a wise advise of you veteran netters: Many precious foul-stained
slides should be re-stained rather than disposed of. What should I do in
order to:

1. Decolor these slides.
2. Restain them with my new MG and G reagents?

Thank you

Agustin Jose Venzano Halliburton
DVM, INTA, Argentina