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.