From: | Tony Henwood |
Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: (02) 9845 3306
Fax: (02) 9845
3318
http://www.histosearch.com/homepages/TonyHenwood/default.html
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-----Original Message-----
From: manal galal [mailto:galalmkh@yahoo.com]
Sent: Wednesday, 4 June 2003 23:45
To: histonet@pathology.swmed.edu
Subject: merosin and emerin on frozen muscle sections.Dear colleagues,My work is on frozen muscle. I am having trouble with merosin and emerin immunostaining on frozen muscle. I wonder if my freezing process is to blame?My freezing procedure is as follows:1-We recieve the specimen immediately after the biopsy is taken.2-We put it in a fridge for 15 min to relax.3- The section is stuck on a pre-frozen chuck with a dab of OCT.4- It is then immediately placed on the quick freeze shelf (-40C) of the cryostat for 15 min.5-Then the specimen is put in the main compartment of the cryostat (-20C) and left there for 20 min. to accomodate to the temperature.6- The sections are then cut at 8 microns and placed on the slides.7- The slides are left to dry overnight at room temperature.By the way we get very good results with Dystrophins and Sarcoglycans, using this method of freezing.Any suggestions??Thank youManal Galal
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