[Histonet] freezing mouse heart tissue

[David Santer]

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