[Coral-List] Technical question about fixation of coral tissues for EM

Szmant, Alina szmanta at uncw.edu
Mon Jul 23 11:53:39 EDT 2007

Hi All:


We will be fixing some coral larvae and tissue for EM later this summer
and would appreciate any advice on good stopping places in the fixation
sequence:  We will be fixing with 2.5 % glut in Millonig's buffer, and
post fix with 2 % osmium in same buffer.  We can't go all the way to
embedding in the field but can take it though the osmium step.  It will
then be a few weeks before the materials can be dehydrated and embedded
in Spurr's.  So the question is:  where is the best place to stop the
process in the field?


a)       after the glutaraldehye (and whether to store in glut sol'n or
in the PO4 buffer?)


b)       after the osmium step (in buffer or 70 % EtOH??)


Thank you for sharing any tips you've developed over the years.  It's
been awhile since I've used these methods and I imagine better protocols
for coral tissues have been worked out.


Alina Szmant


P.S.  EMS has a new, more expensive low viscosity 2 part Spurr's like
medium:  has anyone tried it?  3 X the price but looks much more



Dr. Alina M. Szmant

Coral Reef Research Group

UNCW-Center for Marine Science 

5600 Marvin K. Moss Ln

Wilmington NC 28409

Tel: (910)962-2362 & Fax:  (910)962-2410

Cell:  (910)200-3913

email:  szmanta at uncw.edu

Web Page:  http://people.uncw.edu/szmanta



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