[Histonet] cryosectioning intervertebral disc

[anclg]

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!