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Improved ultrastructure of marine invertebrates using non-toxic buffers

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...

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Detalles Bibliográficos
Autores principales: Montanaro, Jacqueline, Gruber, Daniela, Leisch, Nikolaus
Formato: Online Artículo Texto
Lenguaje:English
Publicado: PeerJ Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4824901/
https://www.ncbi.nlm.nih.gov/pubmed/27069800
http://dx.doi.org/10.7717/peerj.1860
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author Montanaro, Jacqueline
Gruber, Daniela
Leisch, Nikolaus
author_facet Montanaro, Jacqueline
Gruber, Daniela
Leisch, Nikolaus
author_sort Montanaro, Jacqueline
collection PubMed
description 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 MgCl(2)). 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.
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spelling pubmed-48249012016-04-11 Improved ultrastructure of marine invertebrates using non-toxic buffers Montanaro, Jacqueline Gruber, Daniela Leisch, Nikolaus PeerJ Cell Biology 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 MgCl(2)). 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. PeerJ Inc. 2016-03-31 /pmc/articles/PMC4824901/ /pubmed/27069800 http://dx.doi.org/10.7717/peerj.1860 Text en ©2016 Montanaro et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.
spellingShingle Cell Biology
Montanaro, Jacqueline
Gruber, Daniela
Leisch, Nikolaus
Improved ultrastructure of marine invertebrates using non-toxic buffers
title Improved ultrastructure of marine invertebrates using non-toxic buffers
title_full Improved ultrastructure of marine invertebrates using non-toxic buffers
title_fullStr Improved ultrastructure of marine invertebrates using non-toxic buffers
title_full_unstemmed Improved ultrastructure of marine invertebrates using non-toxic buffers
title_short Improved ultrastructure of marine invertebrates using non-toxic buffers
title_sort improved ultrastructure of marine invertebrates using non-toxic buffers
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4824901/
https://www.ncbi.nlm.nih.gov/pubmed/27069800
http://dx.doi.org/10.7717/peerj.1860
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