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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet