[Histonet] RE: EM fixative composition

[Sherwood, Margaret]

We are a research lab, but we use K2 (Karnovsky's fix) as well.  Since
paraformaldehyde is hazardous (and a bit involved to make up), we simply buy the
glutaraldehyde and paraformaldehyde in sealed ampules and make it up in a 0.1M
cacodylate buffer.  We also check the pH (7.4), not the osmolality.  I freeze
aliquots (3ml) and then thaw to use.  Works out well.

Peggy 


Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDR 214)
Massachusetts General Hospital
50 Blossom Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood <@t> partners.org

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of idimitro <@t> mun.ca
Sent: Thursday, December 08, 2011 9:39 AM
To: Ronald.Houston <@t> nationwidechildrens.org; histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: EM fixative composition

Hi Ronnie,

For EM on clinical samples we use Karnovsky's fixative, which is a
glutaraldehyde/paraformaldehyde solution in 0.1M cacodylate buffer. We prepare
our own but you can buy it. 
We found it does the best job in preserving the cell organelles and membranes.
We do not check the osmolality, but we check the pH and adjust accordingly to
7.4. Keep in the fridge.
I hope this helps.

Iliana Dimitrova, B. Tech, MSc, RT
Histology Supervisor
Room 2808
Medical Education and Laboratory Support Services (MELSS)
Faculty of Medicine
Memorial University of Newfoundland
St. John's, NL Canada A1B 3V6
Tel: 709.777.8389
Fax: 709.777.6746





-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Houston, Ronald
Sent: December 8, 2011 10:53 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] EM fixative composition

Trying to get a sense of how people are making up their glutaradelhyde for EM on
clinical specimens.

Is anyone buying it in premade, and if so from whom?
What buffer system are you using? phosphate or cacodylate (or something else)?
Are you checking the osmolality of the fixative before use?

Thanks

Ronnie Houston, MS HT(ASCP)QIHC
Anatomic Pathology Manager
ChildLab, a Division of Nationwide Children's Hospital
www.childlab.com

700 Children's Drive
Columbus, OH 43205
(P) 614-722-5450
(F) 614-722-2899
ronald.houston <@t> nationwidechildrens.org<mailto:ronald.houston <@t> nationwidechildrens
.org>
www.NationwideChildrens.org<http://www.nationwidechildrens.org/>

"One person with passion is better than forty people merely interested."
~ E.M. Forster



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