[Histonet] Bluing

[WILLIAM DESALVO]

Blueing will differentiate and can help crisp the stain. The pH does affect the rate and intensity of color change. You want a slightly alkaline solution to “Blue” your slides. The blueing process will differentiate the normal purplish color of the acid dye, Hematoxylin (pH 2.6-2.9) by changing the color to a blue from the reddish purple. This process produces a better contrast with the usual red counter stains, Eosin and Phloxine, used in the H&E.
 
Common blueing/differentiators I have used are; 0.1% Sodium Bicarbonate (1 g / 1000 ml distilled water), 0.2% Ammonia Water (2 ml / 1000 ml distilled water) and Saturated Lithium Carbonate (1.54 g / 100 ml distilled water). You can also use “tap water”, but be careful about how the water is treated. Rinsing in tap or distilled water also provides a crisper stain because is rinses away the alum. If alum remains, the color will fade with time.

William DeSalvo, B.S., HTL(ASCP)
Production Manager-Anatomic Pathology
Chair, NSH Quality Management Committee
Owner/Consultant, Collaborative Advantage Consulting

 

> Date: Tue, 18 Dec 2012 11:20:26 -0500
> From: turkekul <@t> gmail.com
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Bluing
> 
> Dear Histonetters,
> 
> 
> I wonder what is the purpose of bluing the hematoxylin. Can bluing effect
> the clarity of the nuclear staining? Can one achieve different bluing
> results varying the composition, pH or time of the bluing reagent?
> 
> 
> Mes
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