Tissue Preparation for Vibratome Sectioning

Contributed by Luke Hammond, QBI, The University of Queensland, Australia

Tissue preparation for vibratome sectioning.

  • Fix in 4% paraformaldehyde in 0.1 M PBS pH 7.2-7.4.
  • Remove excess fixative by washing tissues in PBS x 3.
  • Embed in warm 40ºC 4% agarose in distilled water (dH20).
  • Cool tissue blocks to 25ºC or until the agarose solidifies.
  • Trim excess agarose away and attach blocks to metal chuck using cyanoacrylate adhesive.
  • Cut sections to 30-50µm Vibratome sections.
  • Collect sections in 0.1M PBS (add azide 0.05% for long term storage>1 week).

This method is based, with permission, on an original protocol available here.