RE: cutting fixed "sucrose" livers

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From:Cynthia Favara <> (by way of histonet)
To:histonet <>
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		I cut fixed cryopreserved [30% sucrose] mouse brain and
usually have to cut at a lower temperature than fresh frozen. In my limited
experience with this the temp is usually 10C cooler. We have two cryostats
and the temperature setting is different on each one. In essence I would
try colder and not be afraid to go to -30C.
	I also have a higher incidence of sections floating off the slides
during staining[IHC]. I find that poly-l-lysine  is the worst and super
frost the most satisfactory. Good luck!
Cynthia Favara
Hamilton, MT

		-----Original Message-----
		From:	I.H.Straatsburg DIVG
		Sent:	Wednesday, January 13, 1999 12:17 PM
		Subject:	cutting fixed "sucrose" livers

		To start with: thank you for the suggestion to infuse fixed
		with a sucrose gradient before cutting cryostat sections.
		sucrose, the morphology of the sections was awful.
Unfortunately, the
		procedure I am using at the moment requires fixation of the
		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
		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,
		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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