Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution

被引:1384
作者
Chen, Bi-Chang [1 ]
Legant, Wesley R. [1 ]
Wang, Kai [1 ]
Shao, Lin [1 ]
Milkie, Daniel E. [2 ]
Davidson, Michael W. [3 ,4 ]
Janetopoulos, Chris [5 ]
Wu, Xufeng S. [6 ]
Hammer, John A., III [6 ]
Liu, Zhe [1 ]
English, Brian P. [1 ]
Mimori-Kiyosue, Yuko [7 ]
Romero, Daniel P. [8 ]
Ritter, Alex T. [9 ,10 ]
Lippincott-Schwartz, Jennifer [9 ]
Fritz-Laylin, Lillian [11 ]
Mullins, R. Dyche [11 ]
Mitchell, Diana M. [12 ]
Bembenek, Joshua N. [12 ]
Reymann, Anne-Cecile [13 ,14 ]
Boehme, Ralph [13 ,14 ]
Grill, Stephan W. [13 ,14 ]
Wang, Jennifer T. [15 ]
Seydoux, Geraldine [15 ]
Tulu, U. Serdar [16 ]
Kiehart, Daniel P. [16 ]
Betzig, Eric [1 ]
机构
[1] Howard Hughes Med Inst, Ashburn, VA 20147 USA
[2] Coleman Technol Inc, Newtown Sq, PA 19073 USA
[3] Florida State Univ, Natl High Magnet Field Lab, Tallahassee, FL 32310 USA
[4] Florida State Univ, Dept Biol Sci, Tallahassee, FL 32310 USA
[5] Univ Sci, Dept Biol Sci, Philadelphia, PA 19104 USA
[6] NHLBI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA
[7] RIKEN Ctr Dev Biol, Opt Image Anal Unit, Kobe, Hyogo 6500047, Japan
[8] Univ Minnesota, Dept Pharmacol, Minneapolis, MN 55455 USA
[9] NICHHD, Cell Biol & Metab Program, NIH, Bethesda, MD 20892 USA
[10] Addenbrookes Hosp, Cambridge Inst Med Res, Cambridge CB2 0XY, England
[11] Univ Calif San Francisco, Dept Cellular & Mol Pharmacol, San Francisco, CA 94158 USA
[12] Univ Tennessee, Dept Biochem & Cellular & Mol Biol, Knoxville, TN 37996 USA
[13] Max Planck Inst Mol Cell Biol & Genet, D-01307 Dresden, Germany
[14] Max Planck Inst Phys Komplexer Syst, D-01307 Dresden, Germany
[15] Johns Hopkins Sch Med, Ctr Cell Dynam, Howard Hughes Med Inst, Dept Mol Biol & Genet, Baltimore, MD 21205 USA
[16] Duke Univ, Dept Biol, Durham, NC 27708 USA
基金
日本学术振兴会; 欧洲研究理事会;
关键词
FLUORESCENCE MICROSCOPY; HIGH-SPEED; DYNAMICS; CELLS; PROTEIN; ORGANIZATION; CYTOKINESIS; STRATEGIES; MIGRATION; TRACKING;
D O I
10.1126/science.1257998
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
Although fluorescence microscopy provides a crucial window into the physiology of living specimens, many biological processes are too fragile, are too small, or occur too rapidly to see clearly with existing tools. We crafted ultrathin light sheets from two-dimensional optical lattices that allowed us to image three-dimensional (3D) dynamics for hundreds of volumes, often at subsecond intervals, at the diffraction limit and beyond. We applied this to systems spanning four orders of magnitude in space and time, including the diffusion of single transcription factor molecules in stem cell spheroids, the dynamic instability of mitotic microtubules, the immunological synapse, neutrophil motility in a 3D matrix, and embryogenesis in Caenorhabditis elegans and Drosophila melanogaster. The results provide a visceral reminder of the beauty and the complexity of living systems.
引用
收藏
页码:439 / +
页数:13
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