[Coral-List] Optimal fixatives for coral tissues??
Szmant, Alina
szmanta at uncw.edu
Tue Jan 13 14:03:11 EST 2004
Hi Shelley:
I used a fixative called Helly's [also known as Zenkers when made as below with the ZnCl2] for many years and it works well for corals:
1000 gm Zn Cl2
500 gm K dichromate
20 L distilled water
Above is stable indefinitely. For use, add 5 ml of formaldehyde (37 to 40 %) to each 100 ml of Helly's
Decalcification is critical to getting good coral tissue preps. The folowing is better than straight HCl solutions:
0.7 gm/L EDTA
0.008 gm/L KNa tartrate
0.14 gm/L Na tartrate
99.2 ml/L conc. HCl (12N)
Change acid at least twice per day and keep removing crud from decalcifying surfaces (endolithic sponges, etc). Wash for ca. 12 with running tap water (overnight) to remove acid before dehydrating to embed, etc.
Techniques were learned from PaulYevich, US EPA and found in a short technical manual that is not generally available and I am not sure if it was published:
Yevich, P and C. Barszcz. 1981. Preparation of aquatic animals for histopathological examination. US EPA, Environmental Monitoring and Support Laboratory, Cincinnati Ohio USA
*******************************************************************
Dr. Alina M. Szmant
Coral Reef Research Group
Professor of Biology
Center for Marine Science
University of North Carolina at Wilmington
5600 Marvin K. Moss Lane
Wilmington NC 28409-5928
tel (910)962-2362 fax (910)962-2410
email szmanta at uncw.edu
http//people.uncw.edu/szmanta/
******************************************************************
-----Original Message-----
From: coral-list-bounces at coral.aoml.noaa.gov
[mailto:coral-list-bounces at coral.aoml.noaa.gov]On Behalf Of Shelley
Anthony
Sent: Monday, January 12, 2004 7:06 PM
To: coral-list at coral.aoml.noaa.gov
Subject: [Coral-List] Optimal fixatives for coral tissues??
Dear Coral-Listers,
I would very much like to hear from scientists who are dealing with
different fixation techniques for histological work on hard corals. I
have been using standard 5% seawater-buffered formalin, per
recommendation from my supervisor. However, I have had numerous
conflicting suggestions from histologists, lab technicians and other
sources that the following fixatives give better results: 4% formalin,
10% formalin, 70% ethanol, and 73% ethanol, glutaraldehyde, Helley's
fixative, etc. (However, most of these people were not necessarily
working on corals.) The publications I have read also do not seem to
have a standard procedure, or the fixative may be dependent on the
intended usage.
My questions are: Is there an optimal fixative specifically for
hard corals that can be used for several different kinds of
histological/histopathological/microscopic procedures? What kind of
results have you gotten when using different fixatives? I would
appreciate any information you can share. Please reply directly to my
email address. Thanks!
Cheers,
Shelley
--
Shelley L. Anthony <shelleya at gbrmpa.gov.au>
PhD Student, CRC Reef Research Center
School of Marine Biology & Aquaculture
James Cook University
Townsville, QLD 4811 AUSTRALIA
ReefHQ/GBRMPA
Townsville, QLD 4810 AUSTRALIA
(07)4750-0898
================================================================================
If you have received this transmission in error please notify us immediately
by return email and delete all copies. Any unauthorised use, disclosure or
distribution of this email is prohibited.
================================================================================
_______________________________________________
Coral-List mailing list
Coral-List at coral.aoml.noaa.gov
http://coral.aoml.noaa.gov/mailman/listinfo/coral-list
More information about the Coral-List
mailing list