Methods for imaging mammalian mitochondrial morphology: A prospective on MitoGraph.
Methods for imaging mammalian mitochondrial morphology: A prospective on MitoGraph.
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DOI：
10.1016/j.ab.2018.02.022
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发表时间：
2018-07-01
影响因子：
2.9
通讯作者：
Hill RB
中科院分区：
文献类型：
--
作者：
Harwig MC;Viana MP;Egner JM;Harwig JJ;Widlansky ME;Rafelski SM;Hill RB
线粒体的形状多种多样，从小的圆形点状结构到高度互连的网状结构。这种形态多样性对功能很重要，但使量化复杂化。因此，早期的量化工作依赖于各种定性描述，这可以理解地降低了网络的复杂性，导致整个外地的一致性面临挑战。最近应用国家的最先进的计算工具，导致更多的定量方法。这一前瞻性突出了MitoGraph的实施，MitoGraph是一种开源图像分析平台，用于测量最初优化用于酿酒酵母的线粒体形态。在这里，Mitograph在五种不同的哺乳动物细胞类型上进行了评估，所有这些细胞类型都通过MitoGraph分析准确分割。MitoGraph还成功区分了不同的线粒体形态，从完全碎片化到过度细长。还提供了标记线粒体的共聚焦成像的一般建议（使用mito-YFP，MitoTracker染料和免疫染色参数）。MitoGraph的广泛采用将有助于实现长期追求的线粒体形态一致且可重复量化的目标。
Mitochondria are found in a variety of shapes, from small round punctate structures to a highly interconnected web. This morphological diversity is important for function, but complicates quantification. Consequently, early quantification efforts relied on various qualitative descriptors that understandably reduce the complexity of the network leading to challenges in consistency across the field. Recent application of state-of-the-art computational tools have resulted in more quantitative approaches. This prospective highlights the implementation of MitoGraph, an open-source image analysis platform for measuring mitochondrial morphology initially optimized for use with Saccharomyces cerevisiae. Here Mitograph was assessed on five different mammalian cells types, all of which were accurately segmented by MitoGraph analysis. MitoGraph also successfully differentiated between distinct mitochondrial morphologies that ranged from entirely fragmented to hyper-elongated. General recommendations are also provided for confocal imaging of labeled mitochondria (using mito-YFP, MitoTracker dyes and immunostaining parameters). Widespread adoption of MitoGraph will help achieve a long-sought goal of consistent and reproducible quantification of mitochondrial morphology.
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