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Fixation

1. 1.5 ml FSW (0.2 µm filtered seawater) in a 15 ml tube (for small fragments).
2. Cut the sample into approximately 2 cm fragments and place them in the tube.
3. Leave the fragments in the tube with the FSW for 10 min before adding 1M MgCl₂.
4. Add 0.3 ml of 1M MgCl₂ to a single tube, and cover the plate with aluminum foil for 10 min.
5. Remove the FSW from the tube.
6. Add 4% formaldehyde solution into the tube.
   Second option: leave the 1.8 ml of FSW and 1M MgCl₂ solution in the tube and add 0.5 ml of 16% formaldehyde to the tube for a final concentration of 4%.
7. Leave at 4°C overnight or 1–2 hours at room temperature.
8. Wash ×3 with sterile PBS solution.
9. For extended storage, store in 70% sterile ethanol, or leave in PBS for short storage of 2–3 days.