[Histonet] Cryostat sections with small bubbles
kspencer <@t> utmem.edu
Wed Jun 15 10:49:37 CDT 2005
I cut rat brain sections at -18 to -20 and often have to warm the block
face with my finger before each section. I also have better luck if I
cut sections at 10u. These are fresh frozen brains. If I cut them at
20u I get bubbles under the sections. I keep them in the cryostat, and
then fix the sections in cold fix, buffer wash, water wash, then air
dry them in the hood.
When I cut rat brain that has been perfused, I cut 20u floating
sections. They never adhere well to slides, even if I use a hot plate.
I always have bubbles under the section. That is why in this case we
use floating sections.
I hope this is helpful.
On Jun 14, 2005, at 8:39 PM, Katri Tuomala wrote:
> Hi Emily,
> I have never heard of hot plating cryostat sections. You may be
> boiling the moisture under the section forming bubbles or do you air
> dry them well first? Also -35 C seems too cold for brain sections, you
> may be getting some cutting artifacts.
> I'm sure there'll be others with helpful information for you.
> Katri Tuomala
> Hamilton, Ontario, Canada
> ----- Original Message ----- From: "Emily Jane Wiesner-Camm"
> <Emily.Wiesner <@t> medecine.unige.ch>
> To: <histonet <@t> lists.utsouthwestern.edu>
> Sent: Tuesday, June 14, 2005 10:36 AM
> Subject: [Histonet] Cryostat sections with small bubbles
>> Hi All,
>> I was wondering whether anyone can let me know why small bubbles
>> appear underneath my rat brain sections when I cut them on a cryostat
>> (at -35) when they appear perfectly flat before placing them on a
>> hotplate. How they can be avoided?
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
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