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

Farias, DR, Simioni, C, Poltronieri, E, Bouzon, ZL and Macleod, CK ORCID: 0000-0002-0539-6361 2017 , 'Fine-tuning transmission electron microscopy methods to evaluate the cellular architecture of Ulvacean seaweeds (Chlorophyta)' , Micron, vol. 96 , pp. 48-56 , doi: 10.1016/j.micron.2017.02.003.

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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 Type: Article
Authors/Creators:Farias, DR and Simioni, C and Poltronieri, E and Bouzon, ZL and Macleod, CK
Keywords: microscopy, seaweed, TEM protocol, ultrastructure, Ulva lactuca
Journal or Publication Title: Micron
Publisher: Pergamon-Elsevier Science Ltd
ISSN: 0968-4328
DOI / ID Number: 10.1016/j.micron.2017.02.003
Copyright Information:

Copyright 2017 Elsevier Ltd.

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