[Histonet] (no subject)

From:Adi Sabag


I'm a PhD student and recently started working with cryo sections.  
I'm working with s.c tumors that are approximately 8x8 mm, they were fixed with
4% PFA and then either frozen in isopentene over liquid N2 or I used a protocol
of embedding in sucrose and then freezing. In both cases I found that the
optimal temperature for getting nice sections is around 20C. I'm using 3
different sizes of sections- 60, 30 and 10um. The problem is that when I'm
trying to pick up the sample from the stage it adheres to the slide from the
margins toward the center of the tumor and at the very center it does not
adhere. This region remains unstuck to the slides and after a few minutes it
starts breaking and a hole is created in the middle of the tissue which gets
bigger with time. After a while the section is almost useless. 
If anyone have an idea on how to deal with this problem it will be a great



Adi Sabag
Tumor Progression and Angiogenesis laboratory
Medicine Faculty, Technion
Haifa, Israel. 

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