Use of Hepes buffer for microalgal culture media and fixation for electron microscopy

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Abstract
N-2-hydroxyethylpiperazine-N-2′-ethanesulfonic acid (Hepes) has been tested as an alternative to tris-(hydroxymethyl)aminoethane (Tris) for hydrogen-ion buffering of algal cultures. Hepes satisfactorily buffered both freshwater and artificial seawater media and has been used successfully to grow 25 species of microalgae from five classes. Electron microscopic fixations of Tetraselmis striata Butcher (Prasinophyceae) and Amphidinium rhyncocephalum Anissimowa (Dinophyceae) buffered by Hepes have given far superior ultrastructural preservation than fixations buffered by Tris. The substitution of Hepes for Tris allows a defined seawater medium to be used as a fixative vehicle for the first time.