Far-field photostable optical nanoscopy (PHOTON) for real-time super-resolution single-molecular imaging of signaling pathways of single live cells
Far-field photostable optical nanoscopy (PHOTON) for real-time super-resolution single-molecular imaging of signaling pathways of single live cells
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DOI：
10.1039/c2nr11739h
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发表时间：
2012-01-01
期刊：
影响因子：
6.7
通讯作者：
Xu, Xiao-Hong Nancy
中科院分区：
文献类型：
--
作者：
Huang, Tao;Browning, Lauren M.;Xu, Xiao-Hong Nancy
细胞信号通路在细胞功能和有效治疗设计中起着至关重要的作用。不幸的是，细胞信号通路的研究仍然具有巨大的挑战性，因为涉及复杂的级联反应，少数分子足以触发单个细胞的信号反应。在这里，我们报告了远场光稳定光学纳米显微镜（PHOTON）与光稳定单分子纳米粒子光学生物传感器（SMNOBS）的发展，用于在单分子（SM）和纳米（nm）分辨率下实时绘制单个活细胞凋亡信号通路的动态级联。我们对单个配体分子（肿瘤坏死因子α， TNF α）及其在单个活细胞上与其受体（TNFR1）的结合动力学进行了定量成像；实时跟踪其簇的形成和内化以及细胞内信号通路的启动；并以足够的时空分辨率研究单个活细胞的凋亡信号动力学和机制。本研究为单个活细胞凋亡信号通路的复杂实时动态级联和分子机制提供了新的见解。PHOTON提供了卓越的成像和传感能力，SMNOBS提供了无与伦比的生物相容性和光稳定性，能够在SM和nm分辨率下实时探测单个活细胞的信号通路。
Cellular signaling pathways play crucial roles in cellular functions and design of effective therapies. Unfortunately, study of cellular signaling pathways remains formidably challenging because sophisticated cascades are involved, and a few molecules are sufficient to trigger signaling responses of a single cell. Here we report the development of far-field photostable-optical-nanoscopy (PHOTON) with photostable single-molecule-nanoparticle-optical-biosensors (SMNOBS) for mapping dynamic cascades of apoptotic signaling pathways of single live cells in real-time at single-molecule (SM) and nanometer (nm) resolutions. We have quantitatively imaged single ligand molecules (tumor necrosis factor alpha, TNF alpha) and their binding kinetics with their receptors (TNFR1) on single live cells; tracked formation and internalization of their clusters and their initiation of intracellular signaling pathways in real-time; and studied apoptotic signaling dynamics and mechanisms of single live cells with sufficient temporal and spatial resolutions. This study provides new insights into complex real-time dynamic cascades and molecular mechanisms of apoptotic signaling pathways of single live cells. PHOTON provides superior imaging and sensing capabilities and SMNOBS offer unrivaled biocompatibility and photostability, which enable probing of signaling pathways of single live cells in real-time at SM and nm resolutions.