Hello, I am having some issues with the sections lifting off the slide, and also getting good sections. The rat femur is decalcified, and was stored in 30% sucrose & Na Azide solution, then put into 30% sucrose for 3-5 days until embedding in OCT and flash freezing that and putting in to the -80 overnight. Next day I put sample into the cryostat that is set at -21. I f tried different positions on the chuck, diamond, horizontal, and longitudinal. I have tried temps from -19 thru -25, also tried 5um up to 50 um, which is huge I know, but I was told to try that.
I would love any suggestions anybody has for me.