Yes, this fixative is FORMALIN FREE.  I have given the original references
for this fixative, and it is useful for CD or cell surface markers,
particularly in rat and mouse, so you can AVOID formalin altogether.  BD
Pharmingen has confused people badly by calling it Zinc Fixative, which
implys zinc formalin - beware, it is formalin free and they refuse or
ignore requests to call it formalin free Zinc Tris buffer fixative, hey -
maybe they are looking in here!!  Sorry, I have pointed this out many times
to BD Pharm and others.  I do make it up in very pure water, and the pH is
NEVER readjusted after adding the salts, this was in original publications.
     

ZINC TRIS BUFFER FIXATIVE

	0.1M TRIS-HCl (MW 157.6)	15.67g/liter MQ water, adjust pH to 7.4 before
adding salts
	
		Calcium acetate				0.5g
		Zinc acetate					5 g
		Zinc chloride					5 g

	Dissolve all salts in 1 liter 0.1M Tris-HCl, pH adjusted to 7.4.  The pH
is not adjusted 	after salts are added

	FIXATION:	Fix tissues for 24 hours up to 72 hours,  use small tissue
pieces, a 			fixation time study is advisable.

	PROCESSING:
	1 hour per change in 70, 90, 95, and 100% ethanol, 2 changes each
	2 changes xylene (xylene substitute) 1 hour each
	Paraffin 3 changes, 1 hour each
	Embed and cut at 5 um, proceed with staining using Alkaline phosphatase as
substrate

	Nitta et al, Cell Vision, Improved in situ immunodetection of leukocytes
on paraffin 	embedded mouse spleen   4(1):73-80, 1997
	Beckstead J HistochemCytochem 1994 42:1127-1134
	Beckstead J HistochemCytochem 1995 43:345

Additional comments:  I know one of the dry salts is hygroscopic(picks up
water), check label/MSDS for this information.   Store it over silica gel
dessicant.  I never had cloudiness, but do store it working fixative in
refrigerator after making up.  

	
At 10:21 AM 2/26/02 +0200, you wrote:
>HISTONETTERS
>Does anyone know the following solution(fixative),used for 
>immunohistochemistry: 0.1MTris,0.05%calcium acetate,0.5% zinc acetate 
>and 0.5% ZnCl2 ph 7.4.? Problems making up the soln. goes cloudy . 
>Ref paper Infection and Immunity Jan 2002, p303-314, "Plasmidic 
>versus Insertional Cloning etc" I. Me'derle' .Can anyone help  
>or can I use Buffered formalin? Thanks. Marilyn
>**********************************************************************
>MRS MARILYN TYLER
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Gayle Callis
MT,HT,HTL(ASCP)
Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367
406 994-4303 (FAX)