Re-hydrating frozen samples

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From:Ian Montgomery <>
Date:Fri, 9 Jul 1999 12:13:58 +0000
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>Date: Fri, 09 Jul 1999 14:20:45 +0800
>From: Tim Fairchild <>
>Subject: Re-hydrating frozen samples
>To: HistoNet Server <>
>Mime-Version: 1.0
>Hi guys,
>I have just come across a new set of problems with the frozen muscle
>Some of the samples we are using have been stored in the -80 freezer for
>about 2 years now.  One factor we did not take into consideration with
>this storing is 'ice burn', or the dehydration of our muscle samples.
>When I go to cut the muscle, the muscle splinters and basically turns to
>powder, thus not being very stainable!

	'Ice burn', your specimens are freeze dried. Not only have you
freeze dried the specimens but ice crystals will be growing away quite
happily at -80. I keep telling people they should store in liquid nitrogen
but sometimes I'm just f*****g in the wind. I'll be interested in the
replies to your posting.
>I guess 2 options arise, firstly, is there a way to cut the dehydrated
>sections (10um), without getting a splintering effect (i.e. put OCT
>around the muscle to keep it in a nice section)?
>The second option would be to re-hydrate the muscle sample in some way.
>We are going to be using the PAS stain (since freezing muscle sections
>breaks open the sarcopasmic reticulum, calcium is on stand-by to break
>glycogen down at a huge rate as soon as the sample is thawed out) and
>also ATPase stain (boiling will kill the enzyme).  Hence, we would need
>a technique that allows rehydration without having to thaw the muscle
>for too long (well as rapidly as possible anyway), or alternately
>re-hydrate the muscle in a medium which isn't conducive to the breakdown
>of glycogen whilst concurrently not killimg the enzyme.
>Any ideas?
>Thanks in advance
>Timothy J. Fairchild B.Sc. (Hons)
>PhD Candidate
>Co-ordinator for Centre of Athletic Testing
>Department of Human Movement and Exercise Science
>Nedlands, Western Australia 6907
>Telephone: (+61 8) 9380 2793
>Facsimile:  (+61 8) 9380 1039

Dr. Ian Montgomery,
West Medical Building,
University of Glasgow,
G12 8QQ,
Tel: 0141 339 8855 Extn. 6602.
Fax: 0141 330 4100.

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