[Histonet] freezing mouse heart tissue

Merced M Leiker leiker <@t> buffalo.edu
Fri Mar 26 10:18:36 CDT 2010


Wow, a whole book has been written on frozen sections? That's fantastic! 
I've got to get a hold of that!


--On Friday, March 26, 2010 8:45 AM -0500 
Charles.Scouten <@t> leica-microsystems.com wrote:

>
>
> There is a thorough discussion of these issues in Dr. Peters Book, "A
> Practical Guide to Frozen Sections"
>
>
>
>
> Cordially,
>
> Charles W. Scouten, Ph.D
>
> Product Manager, MNL
>
> Biosystems Division
>
>
>
> Leica Biosystems Richmond, Inc.
> 5205 Route 12
> P.O. Box 528
> Richmond, IL 60071
> United States of America
>
> Telephone 630 964 0501
>
> facsimile +1 630 964 0576
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> www.MyNeuroLab.com
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> www.leica-microsystems.com
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> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Malika
> Benatti <malbenatti <@t> googlemail.com>
> Sent: Friday, March 26, 2010 8:33 AM
> To: Merced M Leiker <leiker <@t> buffalo.edu>
> Cc: histonet <@t> lists.utsouthwestern.edu
> Subject: Re: [Histonet] freezing mouse heart tissue
>
>
>
> You may found that excessive amount of saline could cause freezing
> artefacts, so dabe
>
> Try to embed heart sample on cryostat chuck directly, with OCT then
> freeze chuck on dry ice block directly as opposed to using liquid
> nitrogen.
>
> Another option, would be freezing tissue in hexane boiling tube, with
> Acetone/dry ice freezing mixture.
>
> Hope this help.
>
> Malika
>
>
>
> On 26 Mar 2010, at 13:22, Merced M Leiker wrote:
>
>> Hi David,
>>
>> I wouldn't snap-freeze the tissue in the OCT blocks. For one this could
>> cause the blocks to crack; you may have noticed this already in some of
>> your blocks?
>>
>> But you may want to try freezing the OCT block by resting the block in a
>> tray of isopentane (2-methylbutane) that has been frozen over liquid N,
>> as you've already thought of.
>>
>> You can also search the Histonet archives on this topic as it's been
>> discussed before here.
>>
>> Regards,
>> Merced
>>
>>
>> --On Friday, March 26, 2010 8:26 AM +0100 David Santer < etw <@t> gmx.at>
>> wrote:
>>
>>> Hello,
>>>
>>>
>>>
>>> I am currently trying to produce cryosections from mouse heart tissue.
>>> I  already have experience with paraffin-sections and had faced no
>>> major  problems. But with cryosectioning I would ask for your help. You
>>> can get  an idea of our current status at
>>> < http://www.d-cup.at/histo/mouseheart.jpg> this link (Hematoxylin test
>>> stain, not H&E, the whole heart section looks like this) and as you
>>> might  guess I am not satisfied with the quality. Would you call this
>>> freezing  artifacts? Some people suggested that freezing with only LN2
>>> would be not  quick enough and create those ice crystals.
>>>
>>>
>>>
>>> Here is how we prepared the tissue:
>>>
>>> After taking out the hearts from the mice, we flushed them retrogradely
>>> via the aorta with cold sodium solution. Then we cut the hearts in
>>> half,  put them into cryomolds and covered them with OCT. Afterwards
>>> they were  snap-frozen in liquid nitrogen and stored at -80?C.
>>>
>>>
>>>
>>> Do you have an advice or maybe a suitable protocol for me? Would you
>>> recommend 2-methyl butane?
>>>
>>>
>>>
>>> Thank you very much! Greetings from the sunny Vienna!
>>>
>>>
>>>
>>> David
>>>
>>> --
>>>
>>> Mit freundlichen Gr??en
>>>
>>> with kind regards
>>>
>>>
>>>
>>> Dr. David Santer
>>>
>>>
>>>
>>> Ludwig Boltzmann Cluster for Cardiovascular Research
>>>
>>> c/o Core Unit for Biomedical Research
>>>
>>> Waehringer Guertel 18-20 - Leitstelle 1Q
>>>
>>> A-1090 Vienna
>>>
>>> Austria
>>>
>>>
>>>
>>> Website: < http://www.cardiovascular-research.at>
>>> www.cardiovascular-research.at
>>>
>>>
>>>
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>>> Histonet <@t> lists.utsouthwestern.edu
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>>>
>>
>>
>>
>> Merced M Leiker
>> Research Technician III
>> Cardiovascular Medicine
>> 348 Biomedical Research Building
>> State University of New York at Buffalo
>> 3435 Main St, Buffalo, NY 14214 USA
>> leiker <@t> buffalo.edu
>> 716-829-6118 (Ph)
>> 716-829-2665 (Fx)
>>
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Merced M Leiker
Research Technician III
Cardiovascular Medicine
348 Biomedical Research Building
State University of New York at Buffalo
3435 Main St, Buffalo, NY 14214  USA
leiker <@t> buffalo.edu
716-829-6118 (Ph)
716-829-2665 (Fx)

No trees were harmed in the sending of this email.
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