Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes

Sang-Hee Shim, Chenglong Xia, Guisheng Zhong, Hazen P. Babcock, Joshua C. Vaughan, Bo Huang, Xun Wang, Cheng Xu, Guo Qiang Bi, Xiaowei Zhuang

Research output: Contribution to journalArticle

249 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)13978-13983
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume109
Issue number35
DOIs
Publication statusPublished - 2012 Aug 28
Externally publishedYes

Fingerprint

Optical Imaging
Organelles
Membranes
Mitochondria
Mitochondrial Dynamics
Lysosomes
Color
Fluorescence
Cell Membrane

Keywords

  • Diffraction limit
  • Nanoscopy
  • Photoactivation localization microscopy
  • Photoswitchable dye
  • Stochastic optical reconstruction microscopy

ASJC Scopus subject areas

  • General

Cite this

Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes. / Shim, Sang-Hee; Xia, Chenglong; Zhong, Guisheng; Babcock, Hazen P.; Vaughan, Joshua C.; Huang, Bo; Wang, Xun; Xu, Cheng; Bi, Guo Qiang; Zhuang, Xiaowei.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 109, No. 35, 28.08.2012, p. 13978-13983.

Research output: Contribution to journalArticle

Shim, Sang-Hee ; Xia, Chenglong ; Zhong, Guisheng ; Babcock, Hazen P. ; Vaughan, Joshua C. ; Huang, Bo ; Wang, Xun ; Xu, Cheng ; Bi, Guo Qiang ; Zhuang, Xiaowei. / Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes. In: Proceedings of the National Academy of Sciences of the United States of America. 2012 ; Vol. 109, No. 35. pp. 13978-13983.
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