AW: [Histonet] frozens from lung-tissue

[Gudrun Lang]

The reason is more the antipathy paired with unfamiliarity with this
reagens. I tried it with a small amount of liquid nitrogen in a plasticbox,
where I placed the (metal) mold into it, with the tissue and the OCT until
it cracks most times. I thought it would be the fastest way of freezing, but
the result didn't suit. So I am not very ambitious to do it again.

The doctor always speaks about "over-freezing" of the edges and a overall
bad morphology.

Thank you for the hint with the cryobath.

Gudrun Lang
 
-----Ursprüngliche Nachricht-----
Von: Cheryl R. Kerry [mailto:tkngflght <@t> yahoo.com] 
Gesendet: Mittwoch, 21. November 2007 19:12
An: gu.lang <@t> gmx.at; histonet <@t> lists.utsouthwestern.edu
Betreff: RE: [Histonet] frozens from lung-tissue

Gudrun--I suspect this isn't quite what you were looking for but have you
guys tried a cryobath?  It's a little blue countertop 'freezer' unit that
has well of pre-chilled isopentane in it, always cold enough to snap freeze
without the drama of liquid nitrogen.  It is about the size of a big toaster
and is ready to go 24/7 with minimal maintenance (temp chart and periodic
cleaning).  It wasn't overly expensive from what I remember for the great
quality of the end results.  We mounted the tissue on a pre-frozen
OCT-covered chuck, covered it with liquid OCT, lowered it into the cryobath
for about 30-40 second and cut...beautiful, artifact free, duplicable and
fast.

What are the reasons s/he doesn't want the more standard snap freeze
methods?  Is it a time or expense issue?

Cheryl



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gudrun Lang
Sent: Wednesday, November 21, 2007 11:51 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] frozens from lung-tissue

Hi to all,

I need the help of those, who have experience on making good frozens from
lung-tissue. We look for a method for freezing lung-tissue without the usage
of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the
institute is not satisfied with the quality and wants us to do better. Our
usual method is to make a OCT-layer on a chuck, let it freeze, give the
tissue on it, surround with OCT, put the cold weight on it, let it freeze
and make the sections.

We tried putting the tissue in the liquid OCT on the chuck without a layer
and  freezing it in a mold with the chuck on top. I also tried first
freezing the piece solely in liquid nitrogen or isopentan, then putting it
on a chuck. All in all - my boss wasn't happy with it.

 

Any input will be appreciated. Thank you in advance

Gudrun Lang