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Proc Natl Acad Sci U S A. 2012 Aug 28;109(35):13978-83. doi: 10.1073/pnas.1201882109. Epub 2012 Aug 13.

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

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1
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.

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.

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PMID:
22891300
PMCID:
PMC3435176
DOI:
10.1073/pnas.1201882109
[Indexed for MEDLINE]
Free PMC Article
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