Fwd: Re: cryo-preservation

From:"Dr. Ian Montgomery." <ian.montgomery@bio.gla.ac.uk>

<html> Jen,<br> <x-tab>        </x-tab>Following on from a posting several weeks ago, attach the muscle to a <b>cork disc </b> with OTC before freezing. This will save you using a cryostat chuck which in a multi-user facility might be in short supply. Plus, why store a valuable chuck when a wee cork disc can be attached to the chuck many times without the risk of muscle thaw. <br> Ian.<br> <br> <br> <blockquote type=cite class=cite cite>Date: Thu, 25 Jan 2001 10:59:02 +0000<br> From: "t.hacker@har.mrc.ac.uk" <T.Hacker@har.mrc.ac.uk><br> Subject: Re: cryo-preservation<br> To: "Philopena, Jennifer" <jennifer.philopena@canji.com><br> Cc: "'Histonet'" <Histonet@pathology.swmed.edu><br> <br> Date sent:      <x-tab>        </x-tab>Wed, 24 Jan 2001 17:23:59 -0500<br> From:           <x-tab>        </x-tab>"Philopena, Jennifer" <jennifer.philopena@canji.com><br> Subject:        <x-tab>        </x-tab>cryo-preservation<br> To:             <x-tab>        </x-tab>"'Histonet'" <histonet@pathology.swmed.edu><br> <br> > Happy Chinese New Year to all in Histoland.<br> > <br> > I am doing fresh frozen sections of muscle and am getting freezing artifact.<br> <br> Jen, I would avoid any form of "cryo-protection" with muscle as this <br> tends to provide the tissue with enough time to develop ice crystal <br> artefact before freezing (producing those nasty little holes within <br> the fibre). Expose the fibres as best you can (small block) and <br> immerse in pre cooled (LN2) isopentane, preferably on the cryostat <br> chuck (specimen holder). If you attach the tissue to the chuck after <br> freezing avoid any thawing and re-freezing as this will introduce <br> even worse artefact.<br> The above observations are the result of personal experience and a <br> number of frustrating hours attempting to get perfectly preserved <br> muscle fibres.<br> Best of luck/skill,<br> Terry.<br> Terry Hacker,<br> Medical Research Council,<br> Harwell,<br> Didcot,<br> Oxfordshire, OX11 ORD<br> 01235 834393 x360</blockquote> <x-sigsep><p></x-sigsep> <font color="#0000FF">Dr. Ian Montgomery,<br> West Medical Building,<br> University of Glasgow,<br> Glasgow,<br> G12 8QQ.<br> Tel: 0141 339 8855.  Extn:6602.<br> Fax: 0141 330 2923<br> e-mail: ian.montgomery@bio.gla.ac.uk</font></html>
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