Live-cell super-resolved PAINT imaging of piconewton cellular traction forces.

Live-cell super-resolved PAINT imaging of piconewton cellular traction forces.
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DOI:
10.1038/s41592-020-0929-2
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发表时间:
2020-10
期刊:
影响因子:
48
通讯作者:
Salaita K
Salaita K
中科院分区:
生物学1区
文献类型:
--
作者:
Brockman JM;Su H;Blanchard AT;Duan Y;Meyer T;Quach ME;Glazier R;Bazrafshan A;Bender RL;Kellner AV;Ogasawara H;Ma R;Schueder F;Petrich BG;Jungmann R;Li R;Mattheyses AL;Ke Y;Salaita K

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尽管机械力在生物学中发挥着至关重要的作用,但目前尚无以亚 100 nm 分辨率对细胞力进行成像的技术报道。在这里,我们提出张力-PAINT (tPAINT),将分子张力探针与 DNA-PAINT 技术相结合,以约 25 nm 的分辨率绘制 pN 机械事件图。为了进行活细胞动态张力成像,我们设计了可逆探针,其具有神秘的对接位点,只有当探针受到的力超过定义的机械阈值(~7-21 pN)时才会显露出来。此外,我们报告了第二种类型的不可逆 tPAINT 探针,它永久暴露其神秘的对接,从而随着时间的推移整合力的历史,提供改进的空间分辨率以换取时间动态。我们应用两种类型的 tPAINT 探针来绘制活人血小板和小鼠胚胎成纤维细胞中的整合素受体力图。重要的是,tPAINT 揭示了细胞前缘的血小板力与 Arp2/3 复合物成核的动态富含肌动蛋白的环之间的联系。
Despite the vital role of mechanical forces in biology, no technique has been reported to image cellular force with sub-100 nm resolution. Here, we present tension-PAINT (tPAINT), integrating molecular tension probes with the DNA-PAINT technique to map pN mechanical events with ~25 nm resolution. To perform live-cell dynamic tension imaging, we engineered reversible probes with a cryptic docking site revealed only when the probe experiences forces exceeding a defined mechanical threshold (~7–21 pN). Additionally, we report a second type of irreversible tPAINT probe that exposes its cryptic docking permanently and thus integrates force history over time, offering improved spatial resolution in exchange for temporal dynamics. We applied both types of tPAINT probes to map integrin receptor forces in live human platelets and mouse embryonic fibroblasts. Importantly, tPAINT revealed a link between platelet forces at the leading edge of cells and the dynamic actin-rich ring nucleated by the Arp2/3 complex.
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发表时间: 2013-04-10
影响因子: 15
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