title: Improved ultrastructure of marine invertebrates using non-toxic buffers.
abstract: Many marine biology studies depend on field work on ships or remote sampling locations where sophisticated sample preservation techniques (e.g., high-pressure freezing) are often limited or unavailable. Our aim was to optimize the ultrastructural preservation of marine invertebrates, especially when working in the field. To achieve chemically-fixed material of the highest quality, we compared the resulting ultrastructure of gill tissue of the mussel Mytilus edulis when fixed with differently buffered EM fixatives for marine specimens (seawater, cacodylate and phosphate buffer) and a new fixative formulation with the non-toxic PHEM buffer (PIPES, HEPES, EGTA and MgCl2). All buffers were adapted for immersion fixation to form an isotonic fixative in combination with 2.5% glutaraldehyde. We showed that PHEM buffer based fixatives resulted in equal or better ultrastructure preservation when directly compared to routine standard fixatives. These results were also reproducible when extending the PHEM buffered fixative to the fixation of additional different marine invertebrate species, which also displayed excellent ultrastructural detail. We highly recommend the usage of PHEM-buffered fixation for the fixation of marine invertebrates.
authors.affiliation: OCUVAC-Center of Ocular Inflammation and Infection, Laura Bassi Centers of Expertise, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna , Vienna , Austria.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Montanaro
authors.affiliation: Core Facility Cell Imaging and Ultrastructure Research, University of Vienna , Vienna , Austria.
authors.forename: Daniela
authors.initials: D
authors.lastname: Gruber
authors.affiliation: Max Planck Institute for Marine Microbiology, Bremen, Germany; Department of Ecogenomics and Systems Biology, University of Vienna, Vienna, Austria.
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Leisch
keywords: Electron microscopy; Immersion fixation; PHEM buffer