merosin and emerin on frozen muscle sections.
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.
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