SEM Preparation

SEM Preparation

SEM of Squid Tentacle Sucker
  1. Fixation Fixation 1-2 hrs (or overnight at 4C) 2.5% Glutaraldehyde 2.5% Paraformaldehyde in 0.1 M sodium cacodylate buffer (pH 7.4). 
  2. Rinse 3 X 10 min in 0.1M sodium cacodylate buffer, pH 7.4
  3. Post-Fix 1-2 hrs in 1% Osmium tetroxide in water
  4.  Rinse 3 X 5 min in water
  5. Dehydrate
    50% ETOH 5 min
    70% ETOH 5 min
    95% ETOH 5 min
    100% ETOH 10 min
    100% ETOH 10 min
  6. Drying: Critical Point Dry (CPD) or dry using HMDS. 
  7. Mount specimens onto stubs by using conductive carbon tape, silver paint, or colloidal graphite. If you use paint or graphite, it must be dried overnight before going into the Sputter Coater.
  8. Sputter Coating. The thickness you use can vary between 1- 10 nanometers. 
  9. SEM Imaging