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

Shim S.-H.; Xia C.; Zhong G.Babcock H.P.Vaughan J.C.Huang B.Wang X.Xu C.Bi G.-Q.Zhuang X.

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

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

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摘要

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.

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《Proceedings of the National Academy of Sciences of the United States of America》 |2012年第35期|13978-13983|共6页
作者
Shim S.-H.; Xia C.; Zhong G.Babcock H.P.Vaughan J.C.Huang B.Wang X.Xu C.Bi G.-Q.Zhuang X.;
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作者单位

Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, United States;

Hefei National Laboratory for Physical Sciences at the Microscale, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230026, China;

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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
原文格式 PDF
正文语种英语
中图分类自然科学总论;
关键词
Diffraction limit; Nanoscopy; Photoactivation localization microscopy; Photoswitchable dye; Stochastic optical reconstruction microscopy;

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

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