>Date: Fri, 09 Jul 1999 14:20:45 +0800
>From: Tim Fairchild <timf@cyllene.uwa.edu.au>
>Subject: Re-hydrating frozen samples
>To: HistoNet Server <HistoNet@pathology.swmed.edu>
>Mime-Version: 1.0
>
>Hi guys,
>
>I have just come across a new set of problems with the frozen muscle
>sections.
>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!

Tim,
	'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.
Ian.
>
>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
>Tim.
>-----------------------------------------------------------------
>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
>Email: timf@cyllene.uwa.edu.au
>http://www.general.uwa.edu.au/~hmweb/index.htm
>

Dr. Ian Montgomery,
West Medical Building,
University of Glasgow,
Glasgow,
G12 8QQ,
Scotland.
Tel: 0141 339 8855 Extn. 6602.
Fax: 0141 330 4100.
e-mail: ian.montgomery@bio.gla.ac.uk