Hi everyone, I'm trying to cryosection mouse white adipose tissue. The end-goal is to immunostain these sections.
Unfortunately, all of my sections are autofluorescing heavily. My protocol is: 1. Harvest tissue and rinse in PBS 2. Fix for 2 hours in 4% PFA at 4 Celsius 3. Wash 3x with PBS for 5 minutes per wash 4. Incubate in 30% sucrose overnight at 4C 5. Wash with O.C.T. 6. Incubate in O.C.T. overnight at 4C 7. Embed and cryosection 8. Bring to room temp in PBS 9. Check sections under the microscope (no mounting, no staining yet) I thought perhaps I was over-fixing the tissues, so I dropped the 4% PFA fix time to 15 minutes. I don't see any notable difference in the degree of autofluorescence. Thanks in advance for any help you can provide. Frank _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
