Re: [Histonet] Cryostat sections with small bubbles

From:Kathleen Spencer

Hi Emily,

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.

Kathleen Spencer

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
> Katri Tuomala
> Hamilton, Ontario, Canada
> ----- Original Message ----- From: "Emily Jane Wiesner-Camm" 
> To: 
> 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?
>> Thanks,
>> Emily
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