cutting fixed "sucrose" livers

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From:"I.H.Straatsburg DIVG" <I.H.Straatsburg@AMC.UVA.NL> (by way of histonet)
To:histonet <>
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To start with: thank you for the suggestion to infuse fixed tissue
with a sucrose gradient before cutting cryostat sections. Without
sucrose, the morphology of the sections was awful. Unfortunately, the
procedure I am using at the moment requires fixation of the liver
tissue prior to freezing. I would rather do without.

Here comes the problem: the liver and kidney tissue infused with 50%
sucrose in 0.1 M Phosphate buffer (until it sank) is very difficult
to cut in a cryostat (-20 degrees Celsius; 8-10 micrometers
Note: The tissue samples were not impregnated in OCT compound (I take
that is 'Tissue Tec' or something similar) before freezing, but
directly frozen either in liquid N2 or on a cryo-boost-plate in the
cryostat cabinet.

Should I cut a lower temperatures? I have got the impression, the
tissue is too soft, too sticky.
Is the impregnation with OCT compund essential?

I am very curious and thankful for any help!
Dr. I.H. Straatsburg
IWO1-155, Dept. Exp. Surg., Surgical Lab.,
AMC, Amsterdam, NL 1105 AZ
tel. +31.20.5666653

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