Re: [Histonet] wright-giemsa stain on tissue

From:"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

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
_______________________________ 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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