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.

中文翻译：

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线粒体形态和功能：一价两！

已知线粒体的形状和功能是相互关联的；因此，需要将三维 EM 结构分析与功能分析结合起来。细胞色素 c 氧化酶标记是一种在 EM 水平检查线粒体功能的方法。然而，以前应用这种方法的努力有几个问题，包括结果不一致、线粒体超微结构的破坏以及体积 EM 方法缺乏优化。我们使用短固定和微波处理来解决这些问题。我们表明我们的方法提供了一致的细胞色素 c 氧化酶标记，并提高了整个组织体积的标记渗透。我们还量化了线粒体形态指标，包括体积 EM，以表明超微结构未因处理而改变。