I'm having trouble getting consistent good quality fixed sections from mouse retroperitoneal and epididymal fat. I need high quality morphology (intact cell membranes) in order to do computer based cell size measurements from 6µm sections. I fix 1x1 cm tissue in 10% formalin for 4 days, dehydrate to xylene and embed in paraffin. I get very variable result that I belive has to to with processing and not with the microtome cutting. I have tried sections from 4 to 12µm but thick sections tend to be even worse. Sometimes the sections are perfect and less than 1% off the cell membranes are broken, but sometimes the whole set of samples have more than 20% broken cell membranes. Does anyone had a similar experience or have any suggestions on how to best fix adipose tissue in order to get sections without broken cell membranes?
I would be grateful for any help or suggestion.
> Melker Göransson, PhD
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