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Title: Ultrafast single-molecule imaging reveals focal adhesion nano-architecture and molecular dynamics
Authors: Fujiwara, Takahiro K.
Tsunoyama, Taka A.
Takeuchi, Shinji
Kalay, Ziya
Nagai, Yosuke
Kalkbrenner, Thomas
Nemoto, Yuri L.
Chen, Limin H.
Shibata, Akihiro C.E.
Iwasawa, Kokoro
Ritchie, Ken P.
Suzuki, Kenichi G.N.
Kusumi, Akihiro
Author's alias: 藤原, 敬宏
角山, 貴昭
竹内, 信司
根本, 悠宇里
柴田, 明裕
岩沢, こころ
鈴木, 健一
楠見, 明弘
Keywords: Adhesion
Biophysics
Issue Date: 7-Aug-2023
Publisher: Rockefeller University Press
Journal title: Journal of Cell Biology
Volume: 222
Issue: 8
Thesis number: e202110162
Abstract: Using our newly developed ultrafast camera described in the companion paper, we reduced the data acquisition periods required for photoactivation/photoconversion localization microscopy (PALM, using mEos3.2) and direct stochastic reconstruction microscopy (dSTORM, using HMSiR) by a factor of ≈30 compared with standard methods, for much greater view-fields, with localization precisions of 29 and 19 nm, respectively, thus opening up previously inaccessible spatiotemporal scales to cell biology research. Simultaneous two-color PALM-dSTORM and PALM-ultrafast (10 kHz) single fluorescent-molecule imaging-tracking has been realized. They revealed the dynamic nanoorganization of the focal adhesion (FA), leading to the compartmentalized archipelago FA model, consisting of FA-protein islands with broad diversities in size (13–100 nm; mean island diameter ≈30 nm), protein copy numbers, compositions, and stoichiometries, which dot the partitioned fluid membrane (74-nm compartments in the FA vs. 109-nm compartments outside the FA). Integrins are recruited to these islands by hop diffusion. The FA-protein islands form loose ≈320 nm clusters and function as units for recruiting FA proteins.
Description: 細胞膜上の分子がバレエの群舞のように見えてきた: 1蛍光分子の感度で、究極速度で撮像できるカメラを開発. 京都大学プレスリリース. 2023-06-06.
Rights: © 2023 Fujiwara et al.
This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).
URI: http://hdl.handle.net/2433/283284
DOI(Published Version): 10.1083/jcb.202110162
PubMed ID: 37278764
Related Link: https://www.icems.kyoto-u.ac.jp/news/8175/
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