[Histonet] freezing mouse heart tissue

[Merced M Leiker]

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 Grüßen
>
> 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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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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