Re: frozen eyes - my challenge for the week
|From:||Stephanie Moore <firstname.lastname@example.org>|
I don't know exactly what you need to do...will the eyes be fresh or
fixed? Are you using a cryostat or freezing microtome. I have sectioned
fixed rat eye on a cryostat. They are not difficult to cut at all.
After fixing, I sink them in 30% sucrose for cryoprotection. When ready
I crush up some dry ice, set the chuck in it and put down one layer of
the medium. When all but the center is frozen (center being about equal
to the size of the base of the specimen to be cut) I set them so that
the pupil is facing me and embed with tissue tek OCT medium. After I put
the chuck in the cryostat, I trim the edges and slice away. I slice on
the cryostat at -20 CT and -18 OT at 50 microns, then put the slice onto
a subbed slide. I find eye MUCH easier than brain!
I have no idea if this is even helpful. This is what I have done in the
past and it worked for me.
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