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Stephanie,
I cut 7 um section of formalin-fixed, paraffin-embedded rat brain all
the time. I used to trim my blocks into a square or rectangle shape.
Then I read something about trimming in a trapezoid shape (where the top
of the block is the short edge of the trapezoid). I used to have a
problem with my sections coming apart before all the wrinkles were gone,
however, after trying the trapezoid trimming the problem went away.
Also, making ribbons is still no problem.
Since then, someone attempted to explain to me that it's all about
physics and the distribution of forces! Whatever the reason, it's worth
a try.
Sharron Ladd
Dept. of Anatomy
University of South Florida

Hi,
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.

Thanks,

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





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