[Histonet] thawing and refreezing muscle?

[Lewis, Sarah]

I have tried this method as well and it works just as described below.

Sarah E. Lewis HT, ASCP
The Research Institute at
Nationwide Children's Hospital
Center for Gene Therapy
Neuromuscular Division
Rm WA3110
(614)722-2204


 

-----Original Message-----
From: Tony Reilly [mailto:Tony_Reilly <@t> health.qld.gov.au] 
Sent: Tuesday, October 01, 2013 7:15 PM
To: Histonet; Timothy Morken
Cc: jmitchell <@t> neurology.wisc.edu
Subject: Re: [Histonet] thawing and refreezing muscle?

Hi Tim
 
A guy in our histology group gave a presentation debunking the theory that ice crystal artefact is irreversible.  His lab receives frozen specimens from thousands of kilometres away from labs with no histology staff.  If there was ice crystal artefact he defrosts the specimen at room temperature.  Mop the specimen with tissues to remove as much OCT and moisture as possible.  Allow the specimen to remain at room temperature on the bench for 20-30 minutes depending on the size of the specimen to allow evaporation of more moisture.  Refreeze the specimen with OCT and cut sections.  The result was absence of ice crystal artefact but some separation of muscle bundles which does not interfere with staining.  It appears the ice crystals do not damage the fibres just cause separation.  So removing the moisture eliminates the problem.  If you have a lot of muscle perhaps you could separate a small amount to trial the procedure.
 
regards
Tony
 
 
 

Tony Reilly  B.App.Sc. , M.Sc.
Chief Scientist, Anatomical Pathology
Pathology Queensland-PA Laboratory
________________________________________________
Health Services Support Agency | Department of Health
 
Level 1, Building 15,Princess Alexandra Hospital Ipswich Road,WOOLLOONGABBA  Qld4102
Ph: 07 3176 2412
Mob: 0402 139411
Fax: 07 3176 2930
Email: tony_reilly <@t> health.qld.gov.au
Web:  www.health.qld.gov.au/qhcss/
 
 


>>> "Morken, Timothy" <Timothy.Morken <@t> ucsfmedctr.org> 10/2/2013 2:07 am 
>>> >>>
Histonetters, Does anyone have a good method for thawing muscle and refreezing for histochemistry? With good results? We have some bulk-frozen muscle (centimeter thick) and that is what they want to try.

Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center

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