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Fluorescence-Combined Interferometric Scattering Imaging Reveals Nanoscale Dynamic Events of Single Nascent Adhesions in Living Cells
The Journal of Physical Chemistry Letters ( IF 5.7 ) Pub Date : 2020-11-18 , DOI: 10.1021/acs.jpclett.0c02103
Jin-Sung Park 1 , Il-Buem Lee 1 , Hyeon-Min Moon 1 , Jin-Sun Ryu 2 , Sun-Young Kong 3 , Seok-Cheol Hong 1, 4 , Minhaeng Cho 1, 5
Affiliation  

Focal adhesions (FAs) are dynamic protein nanostructures that form mechanical links between cytoskeletal actin fibers and the extracellular matrix. Here, we demonstrate that interferometric scattering (iSCAT) microscopy, a high-speed and time-unlimited imaging technique, can uncover the real-time dynamics of nanoscopic nascent adhesions (NAs). The high sensitivity and stability of the iSCAT signal enabled us to trace the whole life span of each NA spontaneously nucleated under a lamellipodium. Such high-throughput and long-term image data provide a unique opportunity for statistical analysis of adhesion dynamics. Moreover, we directly revealed that FAs play critical roles in both the extrusion of filopodia as nucleation sites on the leading edge and the one-dimensional transport of cargos along cytoskeletal fibers as fiber docking sites. These experimental results show that iSCAT is a sensitive tool for tracking real-time dynamics of nanoscopic objects involved in endogenous and exogenous biological processes in living cells.

中文翻译:

荧光组合干涉散射成像揭示活细胞中单个新生黏附的纳米级动态事件。

粘着斑(FAs)是动态的蛋白质纳米结构,可在细胞骨架肌动蛋白纤维和细胞外基质之间形成机械连接。在这里,我们证明了干涉散射(iSCAT)显微镜是一种高速且不受时间限制的成像技术,可以揭示纳米级新生粘连(NAs)的实时动态。iSCAT信号的高灵敏度和稳定性使我们能够追踪每个在片状脂质体下自发成核的NA的整个寿命。这种高通量和长期的图像数据为粘附动力学的统计分析提供了独特的机会。此外,我们直接揭示,FAs在丝状伪足的挤出(作为前沿的成核位点)和货物沿细胞骨架纤维的一维运输(作为纤维停靠位点)中都发挥着关键作用。
更新日期:2020-12-03
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