[Histonet] Buffer.

Judith L. Williams juditw <@t> u.washington.edu
Thu Nov 13 15:13:21 CST 2008


Hi Ian - I just happen to have a copy of "Fixation of Fish Tissue" by Fournie, Krol, Hawkins- CH 34, Academic Press, 2000. I have the excerpt so I do not know the title of the book.
I quote them- " The osmolality range for mammalian tissues are routinely used. For fish, glutaraldehyde fixatives in cacodylate buffer wihch fall with in the osmolality range for mammalian tissues are routinely used. Karnovsky's fixative (about 2000 mosm) also is often used for marine specimens. The additon of non-electroylytes (sucrose, glucose, dextran) and electroylytes (CaCL2 and NaCl) stabilize osmolarity and decrease cell swelling.
Recommended formula:
2.5% glutaraldehyde in 0.1M Sodium Cacodylate buffer with 0.6 M sucrose
O.2M sodium cacoduylate buffer, pH7.4 .........250.0 ml
glutaraldehyde, 25%  ........................... 50.0 ml
sucrose.........................................100 gm
distilled water  .............................500 gm

Hope this is helpful.  I have good friends who do a lot of fish histology at Gulf Coast Research Laboratory, Univ. of Southern Miss. Ocean Springs, MS.
If you have further questions- contact Nancy-Brown Peterson there.
Judy

On Thu, 13 Nov 2008, Ian Montgomery wrote:

>            What is the recommended EM buffer for sea water fish? I had
> thought of using sea water itself but would cacodylate or one of the
> phosphates be better and at what molarity?
>
> Ian.
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> Dr. Ian Montgomery,
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Judith Williams, PhD, HT(ASCP)
Research Scientist
Department of Comparative Medicine
University of Washington
Seattle, WA 98195





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