Re: Dominici's Stain
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From: | Richard Pitman <Richard.Pitman@wri-tr.wmids.nhs.uk> |
To: | ryan2@niehs.nih.gov |
Reply-To: | |
Content-Type: | text/plain; charset=US-ASCII |
Richard Pitman FIBMS,
Head MLSO,
Dept of Histology, Cytology & Immunology,
Worcester Royal Infirmary NHS Trust
>>> "Ryan.Linda" <ryan2@niehs.nih.gov> 08/09/00 01:50pm >>>
Hello,
Anybody with experience with Dominici's stain? I would appreciate the
staining procedure and/or any information about this old stain.
Hi Linda,
We use this stain on 1 micron GMA sections, for light microscopy, typically on bone marrow aspirates. Method follows, uses barbiturate buffer I notice !
Buffering reduces background staining of GM sections.
Reagents
Stock staining solutions:
A) 0.5 g Eosin Y and 0.5 g Orange G dissolved in 100 ml distilled water.
B) 0.5 g Toluidine Blue dissolved in 100 ml distilled water.
OR
0.5g Toluidine Blue dissolved in 100 ml of Michaelis Veronal acetate buffer, pH 4.
Michaelis Veronal Acetate buffer:
Stock solution
9.714 g Sodium Acetate 3H20
14.714 g Sodium Barbiturate (Veronal)
500 ml distilled water
Working solution
Using pH meter, take 25 ml of stock solution. Add 0.1 N HCL until pH 4 is reached. Dilute to total volume of 100 ml with distilled water. Dissolve 0.5 g Toluidine Blue in this.
Quality control
Procedure/Methodology
1.Place sections in coplin jar filled with solution A for 10 minutes.
2. Rinse in distilled water to remove excess red staining.
3. Place sections in coplin jar filled with solution B for 35-60 seconds.
4. Rinse briefly in distilled water and blot dry.
5. Dry thoroughly on hot plate, rinse in Xylene and mount in Pertex. / DPX
Normal ranges
Nuclei................................blue
Basophilic cytoplasm........shades of blue
Erythrocytes..........................pink
Eosinophil granules.............bright red
Red cell precursors.........shades of pink
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