[Histonet] Oil of Cajeput??

[Lee & Peggy Wenk]

Here's our procedure for Cresyl echt violet (which, by the way, as a dye -
no longer exists. Use cresyl violet acetate):

REAGENTS:

CRESYL ECHT VIOLET STAINING SOLUTION
0.5 g Cresyl echt violet (Cresyl Violet Acetate)
0.18 g Sodium acetate (CH3COONaC3H2O)
500 mL Distilled water
Acetic acid, concentrated (CH3COOH)		1.5 mL (approximately - see
instructions)
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.


PROCEDURE - Cresyl Echt Violet:
1.	Deparaffinize and hydrate slides through graded alcohol to distilled
water.
2.	Place sections in cresyl echt violet solution	1-2 hours (up to
overnight) at room temperature.
3. 	Differentiate in two changes of 95% ethanol until nuclei and Nissl
granules remain violet and the background is nearly colorless. Check
differentiation with the microscope. 1 to 2 drops of acetic acid many be
added to the first alcohol to speed up differentiation.
4.	Dehydrate through absolute ethanol and clear in xylene.
5.	Coverslip with a synthetic mounting media.

RESULTS:
Nissl granules - violet
Nuclei	   - violet
Bacteria, fungus - blue to purple
Cartilage, mast cell granules	- blue to purple
Background - colorless to a very very pale violet-gray
 

PROCEDURAL NOTES:
1.	Use the microscope to differentiate. Differentiations may need to be
repeated several times.
2.	May be used as a counterstain after the Luxol fast blue procedure. 
      a.	AFTER LFB: Start after the LFB has been differentiated and
is still in d. water, but before it is run up through alcohols into xylene.
      b.	CEV STAIN: Since slides are in d. water after LFB, slides
can go immediately into CEV staining solution.
3.	To demonstrate only Nissl substance, continue to differentiate until
the nuclei are colorless. Definitely add the acetic acid to the
differentiating alcohols.
4.	Cresyl echt violet dye powder is no longer available. (German dye
discontinued in the 1940's.) Substitute cresyl violet acetate (no Color
Index number). 

Peggy A. Wenk, HTL(ASCP)SLS
William Beaumont Hospital
Royal Oak, MI 48073

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Monfils,
Paul
Sent: Thursday, February 28, 2008 11:36 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Oil of Cajeput??

I have a request to do a cresyl echt violet stain for nerve cell
differentiation.  The protocol calls for oil of cajeput.  Has anyone used
this?  Know where to get it?  Or, has anyone done this technique and know of
a suitable substitute for this oil?  Thanks.
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