Hi all, Can someone please provide me with more details regarding cryosectioning of mouse spleen and liver tissue?
Currently, I've been fixing my samples in 4% PFA (they are well fixed - I know that will be the first question asked!), and then cryoprotect the tissue in a series of graded sucrose solutions (15% to 30%) until they sink. However, when I go to place the sections on the slide from the cryostat, they look great initially under the microscope, but once they dry they have poor morphology, especially seen in the liver. I've never run into this issue before, with either brain or spinal cord which are significantly more delicate. I've tried cutting the tissue free-floating as well to see if it was how I was placing the sections on the slide, but nothing seems to work. The morphology in the liver is so terribly compromised that I cannot visualize the sinusoids properly. It is baffling that once they go onto the slide they look okay, but 5 minutes later the tissue appears to "separate" from itself. Does anyone work specifically in frozen tissue sections, liver and spleen in particular? If so, would you be able to help me figure out the best way to generate quality specimens? Thank you! Regards, Allyse Mazzarelli _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
