X-Message-Number: 23350
From: "Basie" <>
Subject: Cryopreservation at high pressure 
Date: Sat, 31 Jan 2004 22:36:18 -0500

Abstract This study compared 4 fixation methods of rabbit articular
cartilage. Tibial plateau samples were prepared either by chemical fixation,
with or without microwave enhancement, freezing at ambient pressure followed
by freeze substitution or high pressure freezing methods. Following the
initial preservation, samples were freeze fractured to expose their internal
structure. Microwave enhanced chemical fixation provided preservation of the
morphological structure within seconds. Conventional immersion fixation
required hours to give comparable preservation. Ambient pressure
cryofixation preserved samples in their hydrated stage within seconds and
the subsequent freeze substitution required 10 days. The collagen structure
was qualita-tively as well preserved as by the other methods, but the
chondrocytes showed severe ice crystal damage. The shrinkage of the samples
cryo fixed at ambient pressure was significantly less than conventional or
microwave methods.Cryo preservation at high pressure resulted in matrix
preservation that was presumably closest to the in vivo state.Collagen fiber
structures were difficult to differentiate,which may be due to the presence
of frozen hydrated proteoglycans throughout the tissue. Each technique
available to preserve cartilage has it's advantages and disadvantages and
the methods best suited for the specific research goal must be chosen.

Scanning Microscopy Vol. 13, No. 1, 1999 (Pages 61-69) 0891-703599$5.00+.25

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