Single-organelle tracking by two-photon conversion.
Single-organelle tracking by two-photon conversion.
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DOI：
10.1364/oe.15.002490
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发表时间：
2007-03
期刊：
影响因子：
3.8
通讯作者：
W. Watanabe;T. Shimada;S. Matsunaga;Daisuke Kurihara;K. Fukui;Shin-Ichi Shin-Ichi Arimura-Shin-Ichi;N. Tsutsumi;K. Isobe;K. Itoh
中科院分区：
文献类型：
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作者：
W. Watanabe;T. Shimada;S. Matsunaga;Daisuke Kurihara;K. Fukui;Shin-Ichi Shin-Ichi Arimura-Shin-Ichi;N. Tsutsumi;K. Isobe;K. Itoh
在细胞生物学中，关于细胞内对象及其动态的空间和时间信息对于这些对象的动态分析是必不可少的。一个特定的细胞内对象可以通过表现出明显的光诱导光谱变化的光激活荧光蛋白来区分。在这里，我们报告了一个单一的细胞器的选择性标记和跟踪，通过使用近红外飞秒激光脉冲的光转换荧光蛋白的双光子转换。我们使用Kaede的双光子光转换对活的烟草BY-2细胞中的单个荧光素进行了选择性标记。使用这种技术，我们证明，在植物中，个别线粒体沿着细胞骨架的定向运动是由肌动蛋白丝介导的，而微管是不需要的线粒体的运动。这种单细胞器标记技术使我们能够跟踪单个细胞器的动态，揭示细胞器动态的机制。该技术在直接跟踪选择性细胞和细胞内结构方面具有潜在的应用。
Spatial and temporal information about intracellular objects and their dynamics within a living cell are essential for dynamic analysis of such objects in cell biology. A specific intracellular object can be discriminated by photoactivatable fluorescent proteins that exhibit pronounced light-induced spectral changes. Here, we report on selective labeling and tracking of a single organelle by using two-photon conversion of a photoconvertible fluorescent protein with near-infrared femtosecond laser pulses. We performed selective labeling of a single mitochondrion in a living tobacco BY-2 cell using two-photon photoconversion of Kaede. Using this technique, we demonstrated that, in plants, the directed movement of individual mitochondria along the cytoskeletons was mediated by actin filaments, whereas microtubules were not required for the movement of mitochondria. This single-organelle labeling technique enabled us to track the dynamics of a single organelle, revealing the mechanisms involved in organelle dynamics. The technique has potential application in direct tracking of selective cellular and intracellular structures.