Live-cell imaging and analysis of actin-mediated mitochondrial fission.
Live-cell imaging and analysis of actin-mediated mitochondrial fission.
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DOI：
10.1016/j.xpro.2022.101958
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发表时间：
2023-03-17
期刊：
影响因子：
--
通讯作者：
Shaw, Robin M.
中科院分区：
文献类型：
--
作者：
Shimura, Daisuke;Shaw, Robin M.
目前的方法，如固定细胞成像或单次快照成像，不足以捕捉细胞分裂素介导的线粒体分裂。在这里，我们提出了一个协议，捕捉肌动蛋白介导的线粒体分裂使用高分辨率延时成像。我们描述了从细胞制备和线粒体标记到活细胞成像和最终分析的步骤。这种方法也适用于分析多个细胞因子介导的细胞器事件，如囊泡运输，膜融合，和活细胞中的内吞事件。有关本方案使用和执行的完整详细信息，请参见Shimura等人。（2021年）。线粒体周围肌动蛋白组织的实时成像引起裂变定位肌动蛋白吸引剂GJA 1 - 20 k，识别特定的裂变点高分辨率延时成像和细胞器动力学处理出版商说明：进行任何实验方案都需要遵守当地机构的实验室安全和道德准则。目前的方法，如固定细胞成像或单次快照成像，不足以捕捉细胞分裂素介导的线粒体分裂。在这里，我们提出了一个协议，捕捉肌动蛋白介导的线粒体分裂使用高分辨率延时成像。我们描述了从细胞制备和线粒体标记到活细胞成像和最终分析的步骤。这种方法也适用于分析多个细胞因子介导的细胞器事件，如囊泡运输，膜融合，和活细胞中的内吞事件。
Current approaches, such as fixed-cell imaging or single-snapshot imaging, are insufficient to capture cytoskeleton-mediated mitochondrial fission. Here, we present a protocol to capture actin-mediated mitochondrial fission using high-resolution time-lapse imaging. We describe steps starting from cell preparation and mitochondria labeling through to live-cell imaging and final analysis. This approach is also applicable for analysis of multiple cytoskeleton-mediated organelle events such as vesicle trafficking, membrane fusion, and endocytic events in live cells. For complete details on the use and execution of this protocol, please refer to Shimura et al. (2021). Real-time imaging of actin organization around mitochondria causing fission Localize actin attractant GJA1-20k which identifies the specific point of fission High-resolution time-lapse imaging and processing of organelle dynamics Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Current approaches, such as fixed-cell imaging or single-snapshot imaging, are insufficient to capture cytoskeleton-mediated mitochondrial fission. Here, we present a protocol to capture actin-mediated mitochondrial fission using high-resolution time-lapse imaging. We describe steps starting from cell preparation and mitochondria labeling through to live-cell imaging and final analysis. This approach is also applicable for analysis of multiple cytoskeleton-mediated organelle events such as vesicle trafficking, membrane fusion, and endocytic events in live cells.
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