Re: Dominici's Stain

<< Previous Message | Next Message >>
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






<< Previous Message | Next Message >>