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Fine-tuning transmission electron microscopy methods to evaluate the cellular architecture of Ulvacean seaweeds (Chlorophyta)

Citation

Farias, DR and Simioni, C and Poltronieri, E and Bouzon, ZL and Macleod, CK, Fine-tuning transmission electron microscopy methods to evaluate the cellular architecture of Ulvacean seaweeds (Chlorophyta), Micron, 96 pp. 48-56. ISSN 0968-4328 (2017) [Refereed Article]

Copyright Statement

Copyright 2017 Elsevier Ltd.

DOI: doi:10.1016/j.micron.2017.02.003

Abstract

Chemical fixation is a critical step in the analysis of the ultrastructure of seaweeds because the wrong approach can compromise the ability to distinguish fine-scale cellular composition. Fixation agents, fixation time and type of tissue are important factors to consider for transmission electron microscopy (TEM), and not every protocol is suitable for all cell types. We evaluated a range of fixation agents, post-fixation time and dehydration solutions to determine a TEM protocol for seaweeds in the Family Ulvaceae. We assessed Ulva lactuca using 5 protocols. The level of preservation obtained differed markedly between fixation methods The best result was obtained by fixing the sample with 2.5% glutaraldehyde, 0.05 M sodium cacodylate buffer and 2% paraformaldehyde overnight, and 8 h post-fixation in 1% in osmium tetroxide 1%. This approach and fixation time ensured that the membranes, especially the thylakoid membranes of chloroplasts, remained intact. Ethanol is recommended for dehydration as the use of acetone for dehydration resulted in the collapse of cellular membranes. This new protocol will ensure the ultrastructure of Ulvacean seaweeds can be clearly ascertained in the future.

Item Details

Item Type:Refereed Article
Keywords:microscopy, seaweed, TEM protocol, ultrastructure, Ulva lactuca
Research Division:Environmental Sciences
Research Group:Environmental Science and Management
Research Field:Natural Resource Management
Objective Division:Animal Production and Animal Primary Products
Objective Group:Environmentally Sustainable Animal Production
Objective Field:Management of Solid Waste from Animal Production
Author:Farias, DR (Mrs Daniela Farias)
Author:Macleod, CK (Associate Professor Catriona MacLeod)
ID Code:124275
Year Published:2017
Web of Science® Times Cited:1
Deposited By:Ecology and Biodiversity
Deposited On:2018-02-16
Last Modified:2018-08-28
Downloads:0

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