frozen sections--help!

Janet Vaglia jvaglia at IS2.DAL.CA
Tue Apr 22 09:06:28 EST 1997


I have been working with cryostat (OCT) sectioning on z-fish embryos
younger than 30 ss.  Although I have followed the protocol in the
'Zebrafish Book' (incl. fixing in 4% paraformaldehyde, soaking in
sucrose/PBS, freezing in liquid nitrogen, and doing a quick H & E
stain to view the tissue), as well as variations on this theme, I have had
only minor success.  The internal tissue is almost always broken apart
or slightly misshapen (and in worse cases scattered).  It looks like
shrinkage might be part of the problem, but there is likely something else
going on.  I would greatly appreciate any advice on how to improve the
results of OCT sectioning, or perhaps there is a better alternative.
Unfortunately, plastic sectioning is not an option.  THANK YOU!

cheers,
Janet Vaglia

				====================================

oo       ,.. /'''''\ ..,     _		Janet L. Vaglia
 oo   ,"      >>       "    / |		
 o   /  @        > >    `` /  |		Department of Biology
    ()     (,,,)   >>   ,, \  |		Dalhousie University
      ",              "     \_|		Halifax, Nova Scotia B3H 4J1
        ' ~~~~\,,,/ '"			CANADA
    ~~~~~~~~~~~~~~~~~~~~~~~~~~~~
      ~~~~~~~   ~~~~~~  ~~~~~~		Phone:	(902) 494-3335
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					e-mail: jvaglia at is2.dal.ca 










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