[Histonet] Autofluorescence in murine white adpose tissue cryosections

[Frank Lau]

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