[Histonet] cryosectioning intervertebral disc
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
trying frozen, wax embedding and resin embedding this tisse. Can
help please!! Thanking you all in anticipation of a miracle method.
:- The volume of the disc tissue (mainly annulus) is approx 1cm^3
frozen is isopentane. So far we have only been able to take
annulus perpendicular to the spinal axis (ie transverse sections).
ideally like to take sections of unfixed tissue not only in the
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
tried various slice thicknesses from 10 - 40 microns and heating the
increase the temperature differential
*mounting sections flat - The sections are normally folded or
*tears in sections - we have increased the temperature of the table
to 20 degC
*Flaking when sectioning parallel to layers - no suggestions for
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