RE: [Histonet] FW: Cutting frozen tumour

If your tumors are stored in LN without any cryoprotectant (OCT or Sucrose)
you will probably have trouble cutting them.  I would suggest that the
tumors be cryoprotected before freezing and storing in LN.

-----Original Message-----
[] On Behalf Of Martin S.
Sent: Friday, December 15, 2006 3:09 AM
Subject: [Histonet] FW: Cutting frozen tumour



Hi there,
I'm having some trouble cutting frozen tumour samples. The tumours were
stored in liquid nitrogen and i embedded them in OCT in a dish of
isopentane on dry ice.
After cutting, the sections seem to have a very poor structure - bits of
the tissue have holes and some bits look stretched. There are also some
areas that seem to have high background with all antibodies and the
HRP/DAB. I dont know whether its a problem with embedding/cutting or if
this is just what tumours look like! - I'm used to cutting spleen and
lymph nodes. I've been using a number of different types of tumours
(breast, lung, colon etc) from human and mouse.
Some of the tumours I havent been able to cut at all they just crumble -
I have tried as low as -30oC and as high as -12oC cutting temp.
Any suggestions greatly appreciated.

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