[Histonet] Cresyl echt Violett for Nissl substance

Lee & Peggy Wenk lpwenk <@t> sbcglobal.net
Tue May 7 19:02:32 CDT 2013

Cresyl Echt Violet (CEV) is the pre-WWI name. After factories in Germany 
were bombed, the formulation for this stain was lost (that's the story I 
heard in the 70's).

Closest companies since then have made is cresyl violet acetate (CVA), 
though for many decades, continued to call it CEV. Only recently  (maybe 
last 10 years) have companies been calling it CVA, which is confusing to 
labs that need to order a 25 gram bottle once every 25 years.

Here's how we make it up:
Cresyl echt violet (Cresyl Violet Acetate)        0.5 g
Sodium acetate (CH3COONa3H2O)        0.18 g
Distilled water        500.0 mL
Acetic acid, concentrated (CH3COOH)        1.5 mL (up to)
Dissolve cresyl echt violet and sodium acetate in distilled water. Slowly 
add acetic acid, drop by drop, to solution. Should have a pH of 3.5. If 
solution pH is below 3.5, add more sodium acetate. If solution pH is above 
3.5, add more acetic acid. Filter. Let stand overnight before using. Store 
at room temperature. Stable for months. May be reused until weak.

Stain for 1-2 hours. Differentiate in 2 changes 95% alcohol, until 
background cytoplasm is clear (check with microscope). Run up through 2 
changes 100% reagent alcohol, 2 changes xylene or substitute, coverslip. 
Nissl, RNA and DNA (nuclei) will be purple.

To speed up differentiation, and to pull more out of the DNA/nuclei, add 2 
drops of acetic acid conc. to the first 95% alcohol.

It might be CI # 51180, but I don't have that info at home.

Peggy A. Wenk, HTL(ASCP)SLS
Beaumont Hospital
3601 W. 13 Mile Rd.
Royal Oak, MI 48073

Opinions expressed by me to not represent my place of employment.

-----Original Message----- 
From: Yaskovich, Ruth A (NIH/NIDCR) [E]
Sent: Thursday, May 02, 2013 3:52 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Cresyl echt Violett for Nissl substance

What is everyone using for this stain?
Ruth Yaskovich
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Histonet <@t> lists.utsouthwestern.edu

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