Re: Methyl Green pyronin staining
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From: | Laurence Reilly <laurie.reilly@jcu.edu.au> |
To: | "Sarah Christo" <schristo@cvm.tamu.edu>, <HistoNet@pathology.swmed.edu> |
Reply-To: | |
Date: | Thu, 29 Jul 1999 10:34:34 +1000 |
Content-Type: | text/plain; charset="us-ascii" |
Dear Sarah,
The method we have used came originally from Elias,J.M. (1969)
Stain Technology 44 201 via Arthur Rowlatt at CSIRO in Melbourne, Australia.
Like all good chefs we have altered the method to suit our own needs, which
has been to demonstrate plasma cells.
Carnoy's is the fixative of choice but formalin fixed tissues can be
stained if the ratio of the staining solutions is altered.
Method.
First, the 2% Methyl Green solution must be cleared of violet colours by
extraction with chloroform
1. Take sections to water
2. Stain in Methyl Green-Pyronin solution (freshly made) 3 hrs
If the fixative was formalin If the fixative was Carnoy's
2% aq. Pyronin Y 1.5ml 2% aq. Pyronin Y 3ml
2% aq. Methyl Green 3.0ml 2% aq. Methyl Green 3ml
Tap Water to 50ml Tap Water to 50ml
Townsville tap water is "about neutral"--distilled water dosen't work.
(Buffer solutions may be required)
3. Rinse with tert-butanol
4. Dehydrate in tert-butanol
5. Clear in xylene and mount in DPX (or similar)
Results.
Nuclei -- green-purple
Plasma cell cytoplasm - red
Another reference that I have been given for formalin fixed tissues using a
pH 3.8 is Ahlquist, J. (1972) Stain Technology 47 17, but I haven't tried
this.
Good luck with it. Regards, Laurie.
Mr.Laurie Reilly Ph 07 4781 4468
Physiology & Pharmacology Fax 07 4781 5558
Aust.Inst.of Tropical Vet.& Animal Sc.
James Cook University laurie.reilly@jcu.edu.au
Townsville Qld. 4811
Australia.
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