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Harvard University

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Proc. Natl. Acad. Sci. USA, August 28, 2012 vol. 109 no. 35 13978-13983. Published online before print August 13, 2012, doi: 10.1073/pnas.1201882109 Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes

Abstract

Sang-Hee Shim^a,1, Chenglong Xia^b,1, Guisheng Zhong^a,c, Hazen P. Babcock^a,d, Joshua C. Vaughan^a,c, Bo Huang^a,2, Xun Wang^b, Cheng Xu^b, Guo-Qiang Bi^b,3, and Xiaowei Zhuang^a,c,d,e,3

^aDepartment of Chemistry and Chemical Biology,
^cHoward Hughes Medical Institute,
^dCenter for Brain Science, and
^eDepartment of Physics, Harvard University, Cambridge, MA 02138; and
^bHefei National Laboratory for Physical Sciences at the Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230026, China

Abstract
Imaging membranes in live cells with nanometer-scale resolution promises to reveal ultrastructural dynamics of organelles that are essential for cellular functions. In this work, we identified photoswitchable membrane probes and obtained super-resolution fluorescence images of cellular membranes. We demonstrated the photoswitching capabilities of eight commonly used membrane probes, each specific to the plasma membrane, mitochondria, the endoplasmic recticulum (ER) or lysosomes. These small-molecule probes readily label live cells with high probe densities. Using these probes, we achieved dynamic imaging of specific membrane structures in living cells with 30-60 nm spatial resolution at temporal resolutions down to 1-2 s. Moreover, by using spectrally distinguishable probes, we obtained two-color super-resolution images of mitochondria and the ER. We observed previously obscured details of morphological dynamics of mitochondrial fusion/fission and ER remodeling, as well as heterogeneous membrane diffusivity on neuronal processes.

nanoscopy, diffraction limit, photoswitchable dye, stochastic optical reconstruction microscopy, photoactivation localization microscopy

Footnotes
¹S.-H.S. and C. Xia contributed equally to this work.
²Present address: Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94158.
³To whom correspondence may be addressed.

Author contributions: S.-H.S., J.C.V., B.H., G.-Q.B., and X.Z. designed research; S.-H.S. and C. Xia performed research; S.-H.S., C. Xia, G.Z., H.P.B., B.H., X.W., and C. Xu contributed new reagents/analytic tools; S.-H.S. and C. Xia analyzed data; and S.-H.S., G.-Q.B., and X.Z. wrote the paper.

논문정보

형식Research article
게재일2012년 08월 (BRIC 등록일 )
연구진국외 연구진
분야
피인용횟수60회 이상 인용된 논문 (상위피인용논문 등록일 2015-08-31)

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