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MESAB
Tricaine: 3-amino benzoic acid ethyl ester from Sigma (Cat # A-5040).
Mix in fish safe container with a stir bar:
400 mg tricaine powder
800 mg Na2HPO4 (anhydrous)
100 ml glass distilled water
Adjust to ~pH 7 with a drop at a time of 1N NaOH or 1N HCl if needed but it'sit’s
usually right if you weigh the sodium phosphate carefully and measure the water with a
graduated cylinder.
For storage: Aliquot into 6 x 25 ml fish safe plastic bottles and store at 4C. Label
with date made and use within a couple of weeks.
Dilution for babies: 10 drops in fish water (water in which fish are kept) in a 35 mm
diameter disposable Petri dish. (~40-50 mls of liquid). Embryonic zebrafish can
withstand being in MESAB for hours or until the anesthetic wears off. This will slow
their growth rate, but does not seem to cause abnormal development of structures.
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