[Histonet] wright-giemsa stain on tissue

[John A. Kiernan]

Yes. Such methods have been available for many
years. The older ones are somewhat complicated to
do, and require fixation of the tissue in Helly's
or other mercury-containing mixtures. A simplified
method introduced by R.D.Lillie in 1943 (Public
Health Reports 58: 449-452) and refined in 1944
(J. Tech. Methods Bull. Intl Assoc. Med. Museums
24:43).

If you use an azure-eosin (blood) stain on
sections of fixed tissue it is necessary to adjust
the pH. Methanol-fixed blood smears are typically
stained at pH 6.8, but for formaldehyde-fixed
paraffin sections the pH must be lower, in the
range 4 to 5. 

Detailed instructions are given in Lillie &
Fullmer (1976), p.195-197. Lillie, who was a
pathologist, preferred this type of method to H&E
for routine use in the Technicon staining
equipment of his day. I have used it (manually)
and agree with Lillie that it's a much more
informative staining technique than H&E. The pH is
critical, however, and it's necessary to do tests
in the range 3.5 to 5.5 to find the optimum for
the material you are staining. Lillie & Fullmer
provide some guidance on the relation of pH to
type of fixation. pH 4 is often optimal for tissue
that has been properly fixed (overnight or longer)
in formaldehyde.

For a more recent account, see Wittekind, DH et
al. (1991) The standard Romanowsky-Giemsa stain in
histology. Biotechnic & Histochemistry 66:282-295. 
-- 
-------------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
   kiernan[AT]uwo.ca
   http://publish.uwo.ca/~jkiernan/
   http://instruct.uwo.ca/anatomy/530/index.htm
_______________________________
dgaupp <@t> tulane.edu asked:
> 
> Histonet:
> 
> Has anyone ever heard of staining wright-giemsa stain on paraffin embedded
> tissue(rat bone)?  I've stained in the past blood smears and bone marrow
> smears.
> Just wondering if anyone out there in histoland has ever done so.  I checked out
> histonet archives and did not find any results of paraffin processed tissue
> stained with wright-giemsa.
> 
> Thanks,
> 
> Dina
> 
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