Lookup NU author(s): Julie Faitg,
Professor Doug Turnbull,
Dr Kathryn White,
Dr Amy Vincent
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
Mitochondrial shape and function are known to be linked; therefore, there is a need to combine three‐dimensional EM structural analysis with functional analysis. Cytochrome c oxidase labelling is one approach to examine mitochondrial function at the EM level. However, previous efforts to apply this method have had several issues including inconsistent results, disruption to mitochondrial ultrastructure, and a lack of optimisation for volume EM methods. We have used short fixation and microwave processing to address these issues. We show that our method gives consistent cytochrome c oxidase labelling and improves labelling penetration across tissue volume. We also quantify mitochondrial morphology metrics, including in volume EM, to show that ultrastructure is unaltered by the processing. This work represents a technical advance that allows the correlation of mitochondrial function and morphology with greater resolution and volume than has previously been feasible.
Author(s): Faitg J, Davey T, Turnbull DM, White K, Vincent AE
Publication type: Article
Publication status: Published
Journal: Journal of Microscopy
Pages: epub ahead of print
Online publication date: 11/04/2020
Acceptance date: 27/03/2020
Date deposited: 24/04/2020
ISSN (print): 0022-2720
ISSN (electronic): 1365-2818
Publisher: Wiley-Blackwell Publishing Ltd.
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