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

doi:10.1073/pnas.1201882109

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 journal › Article › peer-review

375 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 language	English
Pages (from-to)	13978-13983
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	109
Issue number	35
DOIs	https://doi.org/10.1073/pnas.1201882109
Publication status	Published - 2012 Aug 28
Externally published	Yes

Keywords

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

ASJC Scopus subject areas

General

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10.1073/pnas.1201882109

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Shim, S. H., Xia, C., Zhong, G., Babcock, H. P., Vaughan, J. C., Huang, B., Wang, X., Xu, C., Bi, G. Q., & Zhuang, X. (2012). Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes. Proceedings of the National Academy of Sciences of the United States of America, 109(35), 13978-13983. https://doi.org/10.1073/pnas.1201882109

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title = "Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes",

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.",

keywords = "Diffraction limit, Nanoscopy, Photoactivation localization microscopy, Photoswitchable dye, Stochastic optical reconstruction microscopy",

author = "Shim, {Sang Hee} and Chenglong Xia and Guisheng Zhong and Babcock, {Hazen P.} and Vaughan, {Joshua C.} and Bo Huang and Xun Wang and Cheng Xu and Bi, {Guo Qiang} and Xiaowei Zhuang",

year = "2012",

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doi = "10.1073/pnas.1201882109",

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AU - Shim, Sang Hee

AU - Xia, Chenglong

AU - Zhong, Guisheng

AU - Babcock, Hazen P.

AU - Vaughan, Joshua C.

AU - Huang, Bo

AU - Wang, Xun

AU - Xu, Cheng

AU - Bi, Guo Qiang

AU - Zhuang, Xiaowei

PY - 2012/8/28

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AB - 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.

KW - Diffraction limit

KW - Nanoscopy

KW - Photoactivation localization microscopy

KW - Photoswitchable dye

KW - Stochastic optical reconstruction microscopy

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JF - Proceedings of the National Academy of Sciences of the United States of America

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ER -

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

Abstract

Keywords

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