Direct observation of motor protein stepping in living cells using MINFLUX

被引:71
|
作者
Deguchi, Takahiro [1 ]
Iwanski, Malina K. [2 ]
Schentarra, Eva-Maria [1 ,3 ]
Heidebrecht, Christopher [1 ,3 ]
Schmidt, Lisa [1 ,3 ]
Heck, Jennifer [4 ]
Weihs, Tobias [5 ]
Schnorrenberg, Sebastian [6 ]
Hoess, Philipp [1 ]
Liu, Sheng [1 ,7 ]
Chevyreva, Veronika [1 ,8 ]
Noh, Kyung-Min [4 ]
Kapitein, Lukas C.
Ries, Jonas [1 ]
机构
[1] European Mol Biol Lab, Cell Biol & Biophys Unit, Heidelberg, Germany
[2] Univ Utrecht, Fac Sci, Dept Biol, Cell Biol Neurobiol & Biophys, Utrecht, Netherlands
[3] Heidelberg Univ, Fac Biosci, Heidelberg, Germany
[4] European Mol Biol Lab, Genome Biol Unit, Heidelberg, Germany
[5] Abberior Instruments GmbH, Gottingen, Germany
[6] European Mol Biol Lab, EMBL Imaging Ctr, Heidelberg, Germany
[7] Univ New Mexico, Dept Phys & Astron, Albuquerque, NM USA
[8] FIRC Inst Mol Oncol, Milan, Italy
基金
欧盟地平线“2020”; 欧洲研究理事会;
关键词
TRACKING; RESOLUTION;
D O I
10.1126/science.ade2676
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Dynamic measurements of molecular machines can provide invaluable insights into their mechanism, but these measurements have been challenging in living cells. Here, we developed live-cell tracking of single fluorophores with nanometer spatial and millisecond temporal resolution in two and three dimensions using the recently introduced super-resolution technique MINFLUX. Using this approach, we resolved the precise stepping motion of the motor protein kinesin-1 as it walked on microtubules in living cells. Nanoscopic tracking of motors walking on the microtubules of fixed cells also enabled us to resolve the architecture of the microtubule cytoskeleton with protofilament resolution.
引用
收藏
页码:1010 / 1015
页数:6
相关论文
共 48 条
  • [41] Visualization of Lipid Droplets in Living Cells and Fatty Livers of Mice Based on the Fluorescence of π-Extended Coumarin Using Fluorescence Lifetime Imaging Microscopy
    Yoshihara, Toshitada
    Maruyama, Ryo
    Shiozaki, Shuichi
    Yamamoto, Koji
    Kato, Shin-ichiro
    Nakamura, Yosuke
    Tobita, Seiji
    ANALYTICAL CHEMISTRY, 2020, 92 (07) : 4996 - 5003
  • [42] Dynamic nano-imaging of label-free living cells using electron beam excitation-assisted optical microscope
    Fukuta, Masahiro
    Kanamori, Satoshi
    Furukawa, Taichi
    Nawa, Yasunori
    Inami, Wataru
    Lin, Sheng
    Kawata, Yoshimasa
    Terakawa, Susumu
    SCIENTIFIC REPORTS, 2015, 5
  • [43] Visualizing Protein-DNA Interactions in Live Bacterial Cells Using Photoactivated Single-molecule Tracking
    Uphoff, Stephan
    Sherratt, David J.
    Kapanidis, Achillefs N.
    JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2014, (85):
  • [44] Efficacy and Durability in Direct Labeling of Mesenchymal Stem Cells Using Ultrasmall Superparamagnetic Iron Oxide Nanoparticles with Organosilica, Dextran, and PEG Coatings
    Wang, Yi-Xiang J.
    Quercy-Jouvet, Thibault
    Wang, Hao-Hao
    Li, Ak-Wai
    Chak, Chun-Pong
    Xuan, Shouhu
    Shi, Lin
    Wang, De-Feng
    Lee, Siu-Fung
    Leung, Ping-Chung
    Lau, Clara B. S.
    Fung, Kwok-Pui
    Leung, Ken Cham-Fai
    MATERIALS, 2011, 4 (04): : 703 - 715
  • [45] Efficient electrooxidation of ammonia using platinum supported on titania/sulfur-doped onion like carbon nanoparticles in direct ammonia fuel cells
    Sikeyi, Ludwe L.
    Bila, Laercia R.
    Ntuli, Themba D.
    Selepe, Cyril T.
    Maxakato, Nobanathi W.
    Coville, Neil J.
    Maubane-Nkadimeng, Manoko S.
    DIAMOND AND RELATED MATERIALS, 2023, 132
  • [46] Three-Dimensional Super-Resolution Imaging of the Midplane Protein FtsZ in Live Caulobacter crescentus Cells Using Astigmatism
    Biteen, Julie S.
    Goley, Erin D.
    Shapiro, Lucy
    Moerner, W. E.
    CHEMPHYSCHEM, 2012, 13 (04) : 1007 - 1012
  • [47] Analysis of Her2/neu membrane protein clusters in different types of breast cancer cells using localization microscopy
    Kaufmann, R.
    Mueller, P.
    Hildenbrand, G.
    Hausmann, M.
    Cremer, C.
    JOURNAL OF MICROSCOPY, 2011, 242 (01) : 46 - 54
  • [48] Multimodal Imaging of Living Cells with Multiplex Coherent Anti-stokes Raman Scattering (CARS), Third-order Sum Frequency Generation (TSFG) and Two-photon Excitation Fluorescence (TPEF) Using a Nanosecond White-light Laser Source
    Segawa, Hiroki
    Okuno, Masanari
    Leproux, Philippe
    Couderc, Vincent
    Ozawa, Takeaki
    Kano, Hideaki
    ANALYTICAL SCIENCES, 2015, 31 (04) : 299 - 305