[Histonet] Cryostat sections with small bubbles

[Kathleen Spencer]

Yes Alan, but when I put my finger on the block face to warm it, there 
is a kimwipe or gauze between my finger and the tissue.  It works 
really well, so I guess the Ark is still around.
As the owner of a company that sells cryostats, you need to be aware 
that not everyone has or can obtain such devices.
Many of us, especially in research, work with old and crippled 
equipment, thus we need all the help we can get from our fellow 
technicians.

Kathleen Spencer, HT ASCP
Lab Manager/LCM Supervisor
UTHSC



On Jun 16, 2005, at 5:11 AM, Alan Bright wrote:

> Emily,
>
> Basically you need to section brain at -8 to -12ºC as colder 
> temperatures will cause what you are getting plus cracking. However to 
> achieve good quality sections at these temperatures you will need to 
> maintain the knife and anti-roll temperatures at -20ºC or lower to 
> allow the section to slide in between the knife and the anti-roll 
> plate unimpeded. Which is what Kathleen is doing when she warms the 
> block face with her finger, a technique that went out with the Ark and 
> should not be used as firstly you could have an infection issue and 
> secondly it is not a very scientific approach in this day and age when 
> there are devices fitted to some cryostats such as independent 
> specimen temperature control to perform this task with full and 
> repeatable control and of most importance, ease.
>
> Best Regards
>
> Alan Bright
>
> Bright Instrument Co.Ltd.
> St Margaret's Way
> Huntingdon
> Cambridgeshire
> PE29 6EU
> England
>
> Tel No:+44 (0)1480 454528
> Fax No:+44 (0)1480 456031
> Email: abright <@t> brightinstruments.com
> Web Site: www.brightinstruments.com
>
>
>
> -----Original Message-----
> From: Kathleen Spencer [mailto:kspencer <@t> utmem.edu]
> Sent: 15 June 2005 16:50
> To: Katri Tuomala
> Cc: histonet <@t> lists.utsouthwestern.edu; Emily Jane Wiesner-Camm
> Subject: Re: [Histonet] Cryostat sections with small bubbles
>
>
> 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"
>> <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?
>>> Thanks,
>>> Emily
>>>
>>>
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>>
>>
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