[Histonet] cryosectioning intervertebral disc
Hi everyone,
I`m posting this on behalf of a collegue who wondered if I could
suggest anything to help, but I have had limited success myself
despite
trying frozen, wax embedding and resin embedding this tisse. Can
anyone
help please!! Thanking you all in anticipation of a miracle method.
Chris.
:- The volume of the disc tissue (mainly annulus) is approx 1cm^3
snap
frozen is isopentane. So far we have only been able to take
sections
of the
annulus perpendicular to the spinal axis (ie transverse sections).
I
would
ideally like to take sections of unfixed tissue not only in the
transverse
plane but also radial sections and sections parallel to layers.
The problems we are experiencing are:
*mounting sections to slide - we are currently using APES coated
slides
and have
tried various slice thicknesses from 10 - 40 microns and heating the
slides to
increase the temperature differential
*mounting sections flat - The sections are normally folded or
ruffled
*tears in sections - we have increased the temperature of the table
from 15degC
to 20 degC
*Flaking when sectioning parallel to layers - no suggestions for
this
one yet!
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