Re: FW: brain section blowout
|From:||Tara Miller <email@example.com>|
Another possible problem is the type of paraffin used to embed the brain.
The melting temp may be very low. I know that one place I worked we
accidently ordered the wrong temp paraffin and even though our water baths
were set at 39C, our ribbons started to separate on the water very quickly
(both the sections from each other and the paraffin from the tissue).
Everyone was going crazy for a few hours until I got up and looked at the
bags that the paraffin pellets came in and noticed the printing was a
different color. Aha! Low melting point paraffin.
This is just one more thing to rule out. Good luck.
Tara Oakes, HT
Central Dupage Hospital
>From: Stephanie Rodriguez <firstname.lastname@example.org>
>Subject: FW: brain section blowout
>Date: Fri, 25 May 2001 08:13:15 -0700
>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.
>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.
>Research Technologist III
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