[Histonet] liver cryosections
algranth <@t> u.arizona.edu
Tue Jul 5 12:10:50 CDT 2005
I do many frozen sections on liver tissue and have found that the manner in
which the tissue is frozen makes a huge difference in the way it cuts.
Check to make sure that your samples are being frozen properly. There have
been numerous discussions on histonet about the proper way to freeze tissue
and you can find them in the histonet archives. I have found that if your
tissue looks chalky and whitish then it may have been left in whatever
froze the tissue too long. This makes for real fly away shattered sections
and sometimes cracks the OCT which makes for another nightmare.
Next, I cut my liver frozens at -18 to -16 degrees C. Before taking the
section I often have to rub my thumb gently over the surface of the tissue
(wearing gloves) to slightly warm the surface. This helps to produce a
section that is not shattered. Sometimes a puff of warm air helps too.
Breathe gently on the tissue as you tease down the section and it doesn't
want to fly away.
At 11:45 AM 7/5/2005 -0400, you wrote:
>The block is too cold. -20 degrees is an appropriate temperature for
>sectioning many tissues, but some tissues require a lower temperature and
>others require a higher temperature. For liver, try -15 degrees. For some
>samples you may have to go as high as -12 degrees.
> > ----------
> > From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Till,
> > Renee
> > Sent: Tuesday, July 5, 2005 8:39 AM
> > To: histonet <@t> lists.utsouthwestern.edu
> > Subject: [Histonet] liver cryosections
> > Can anyone offer some advice on cutting liver cryosections? My every
> > attempt produces shredded tissues. I know in paraffin embedding that
> > means they are dry and I would soak it in ice water before I cut, but
> > what do you do with cryosections? I'm pretty sure it's not my cutting
> > technique, though I am fairly new at it. I've adjusted the roll plate
> > and the vacume window. And I've tried it without the roll plate, though
> > as we are just starting out with our cryostat I don't have any good
> > brushes for pulling the section. Any ideas? Maybe they did not have
> > enough time to come down from -80 to -20? I left them in for about 2
> > hours.
> > Renee'
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