RE: Zamboni Solution

<< Previous Message | Next Message >>
From:"Sebree Linda A." <>
To:"'Hall, Phil'" <>
Date:Fri, 18 Jun 1999 07:58:31 -0500

Hi Phil,

Here is the recipe for Zamboni's fixative out of "Theory and Practice of
Histotechnology" by Sheehan and Hrapchak.

Add 20 gm of paraformaldehyde to 150 ml of double-filtered, saturated
solution of picric acid.  Heat solution to 60 degrees C (waterbath or oven)
for 2 hours to dissociate paraformaldehyde into formaldehyde.  Add 2.52%
sodium hydroxide (in water) dropwise to alkalize.  The solution should be
clear.  Filter and allow to cool.  Bring solution to a volume of 1 liter
with phosphate buffer.  The PAF fixative should have a final pH of 7.3 and
an osmolarity of 900 mOsm.  (The range of osmolarity can be adjusted from
600 to 900 to give satisfactory results with tissue fixation.  Tests should
be made to determine what is satisfactory for your particular work.)

Phosphate buffer for Zamboni's fixative:

3.31 gm of NaH2PO4-H20 (monobasic sodium phosphate)
33.77 gm of Na2HPO4-7HsO (dibasic sodium phosphate)
17.88 gm of Na2HPO4

Dissolve salts in 1 liter distilled water.

Good Luck,

Linda Sebree, HT
University of Wisconsin Hospital & Clinics
Department of Laboratory Medicine
IHC/ISH Laboratory
600 Highland Ave.
Madison, WI  53792-2472

Phone:  (608)265-6596
FAX:  (608)263-1568

> -----Original Message-----
> From:	Hall, Phil []
> Sent:	Friday, June 18, 1999 3:51 AM
> To:	'Histonet'
> Subject:	Zamboni Solution
> I am trying a method for NADPH Diaphorase found in the literature.  The
> first step given is to fix in Zamboni solution.  Does anyone have the
> formula to make this up?
> Many thanks
> Phil Hall
> Paediatric Pathology
> St Michael's Hospital
> UK

<< Previous Message | Next Message >>