[Histonet] re embedding of small artery
carl.hobbs <@t> kcl.ac.uk
Mon Sep 13 13:02:14 CDT 2004
In my experienc, with fixed tissues it is easy to embed using agarose, then
to re-orient the tissue for frozen/pwax sectioning: Dissolve 1% agarose( aq)
and when about 50C place the tissue in the agarose, in a bigger mould. It
will set quickly.....better if placed at 4C for a while. Remove the set
agarose and cut away excess agarose so you have the tissue in the
orientation of your choice. Then process to pwax. Or, place in OCT for a
couple of hours on a rotator. Then embed in an OCT filled mould in the
orientation you need and....freeze. If you are doing fresh tissue, get the
melted, cooling agarose into a mould , place the tissue and cool quickly.
Immediately place in OCT, swirl gently for a minute, then freeze. Play
around with control material until you are confident of the technique and to
reassure yourself that the elevated temp does not adversely afffect your
tissue. You can also do gelatin/vibratome sections of the fixed tissue, if
you have a vibratome.
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