Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics
Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics
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DOI：
10.1038/nmeth.1202
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发表时间：
2008-05-01
期刊：
影响因子：
48
通讯作者：
Betzig, Eric
中科院分区：
文献类型：
--
作者：
Shroff, Hari;Galbraith, Catherine G.;Betzig, Eric
我们展示了使用光激活定位显微镜（PALM）的活细胞超分辨率成像。使用耐光细胞系，结合PALM的高分辨率和分子灵敏度，使我们能够在生理条件下研究活细胞中单个粘附复合物（ACs）内的纳米级动态，时间长达25分钟，其中一半的时间用于收集空间分辨率降至60 nm，帧速率短至25 s的PALM图像。我们可视化了AC的形成，并测量了每个AC进化过程中单个paxillin分子的分数增益和损失。通过观察各种各样的纳米级动力学，活细胞PALM提供了在细胞过程的起始、成熟和溶解过程中的分子组装的见解。
We demonstrate live-cell super-resolution imaging using photoactivated localization microscopy (PALM). The use of photon-tolerant cell lines in combination with the high resolution and molecular sensitivity of PALM permitted us to investigate the nanoscale dynamics within individual adhesion complexes (ACs) in living cells under physiological conditions for as long as 25 min, with half of the time spent collecting the PALM images at spatial resolutions down to similar to 60 nm and frame rates as short as 25 s. We visualized the formation of ACs and measured the fractional gain and loss of individual paxillin molecules as each AC evolved. By allowing observation of a wide variety of nanoscale dynamics, live-cell PALM provides insights into molecular assembly during the initiation, maturation and dissolution of cellular processes.