Latin American Journal of Aquatic Research, 41 (5) November 2013
Staining protocol for the histological study of sea anemones (Anthozoa: Actiniaria) with recommendations for anesthesia and fixation of specimens.
Many of the characteristics used in sea anemone taxonomy can only be examined through histological sections. Since there is no standardized procedure for this purpose, various anesthesia and fixation techniques applied to specimens of the intertidal species Anthopleura hermaphroditica and Bunodactis hermafroditica are discussed. Additionally, further modifications are proposed to the Masson\'s trichrome method according to the results obtained on these species. The combined effect of the short application of menthol crystals, together with small doses of MgCl2 were the most satisfactory anesthetics for maintaining the specimens expanded. The best preparations were obtained from samples fixed for several months in 8% seawater formalin; however, in order to achieve a good differentiation of the tissue, mordanting the samples with Bouin\'s fixative was necessary. Besides being a fast method, the modified Masson\'s trichrome gives very good contrasts between the epithelia and the mesoglea, and allows controlling the timing of differentiation during staining. The present paper includes suggestions and precautions and thus offers practical help for the histological study of sea anemones.
Author: Carlos Spano & Verónica Flores

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