Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Cell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht University, Utrecht, Netherlands.
Science. 2023 Mar 10;379(6636):1010-1015. doi: 10.1126/science.ade2676. Epub 2023 Mar 9.
Dynamic measurements of molecular machines can provide invaluable insights into their mechanism, but these measurements have been challenging in living cells. Here, we developed live-cell tracking of single fluorophores with nanometer spatial and millisecond temporal resolution in two and three dimensions using the recently introduced super-resolution technique MINFLUX. Using this approach, we resolved the precise stepping motion of the motor protein kinesin-1 as it walked on microtubules in living cells. Nanoscopic tracking of motors walking on the microtubules of fixed cells also enabled us to resolve the architecture of the microtubule cytoskeleton with protofilament resolution.
动态测量分子机器可以为其机制提供宝贵的见解，但这些测量在活细胞中一直具有挑战性。在这里，我们使用最近引入的超分辨率技术 MINFLUX 在二维和三维空间中以纳米级空间和毫秒级时间分辨率开发了单个荧光团的活细胞跟踪。使用这种方法，我们解析了马达蛋白 kinesin-1 在活细胞中的微管上行走时的精确步进运动。在固定细胞的微管上行走的马达的纳米级跟踪也使我们能够解析微管细胞骨架的结构，达到原丝分辨率。
