FW: brain section blowout

From:Stephanie Rodriguez <srodriguez@primalinc.com>


I know there was a discussion recently about tissue coming apart on the
water bath, but I am still have trouble...

I am cutting 5um sections of formalin-fixed, paraffin-embedded mouse brain.
When I float my ribbons out on my water bath, the sections rapidly come
apart before I have a chance to get them on a slide.

I've tried turning the temp down on my water bath, from 42C to 40C, which
didn't change anything, then down to 35C, which, while it gave me a little
more time, made it almost impossible to get all the wrinkles out of my
sections, so I think that might not be warm enough!

Could it be that the tissue is not adequately fixed? or perhaps poorly
processed?  I fixed the whole mouse brains in 10% NBF for 8hrs at RT, then
72hrs at 4C, then sent them to the contract lab that does our processing.  I
used to work in that lab, and I know their processing schedule is pretty
standard, but perhaps insufficient for these samples?

Any suggestions would be greatly appreciated.


Stephanie Rodriguez
Research Technologist III
Primal, Inc.
Seattle, WA

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