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  STED-FLCS: An advanced tool to reveal spatiotemporal heterogeneity of molecular membrane dynamics.

Vicidomini, G., Ta, H., Honigmann, A., Müller, V., Clausen, M. P., Waithe, D., et al. (2015). STED-FLCS: An advanced tool to reveal spatiotemporal heterogeneity of molecular membrane dynamics. Nano Letters, 15(9), 5912-5918. doi:10.1021/acs.nanolett.5b02001.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-8060-5 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-6006-5
Genre: Journal Article

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 Creators:
Vicidomini, G.1, Author              
Ta, H.1, Author              
Honigmann, A.1, Author              
Müller, V.1, Author              
Clausen, M. P., Author
Waithe, D., Author
Galiani, S., Author
Sezgin, E., Author
Diaspro, A., Author
Hell, S. W.1, Author              
Eggeling, C.1, Author              
Affiliations:
1Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              

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Free keywords: Super-resolved microscopy; stimulated-emission-depletion microscopy; time-resolved; fluorescence-correlation spectroscopy; time-correlated single-photon counting
 Abstract: Heterogeneous diffusion dynamics of molecules play an important role in many cellular signaling events, such as of lipids in plasma membrane bioactivity. However, these dynamics can often only be visualized by single-molecule and super-resolution optical microscopy techniques. Using fluorescence lifetime correlation spectroscopy (FLCS, an extension of fluorescence correlation spectroscopy, FCS) on a super-resolution stimulated emission depletion (STED) microscope, we here extend previous observations of nanoscale lipid dynamics in the plasma membrane of living mammalian cells. STED-FLCS allows an improved determination of spatiotemporal heterogeneity in molecular diffusion and interaction dynamics via a novel gated detection scheme, as demonstrated by a comparison between STED-FLCS and previous conventional STED-FCS recordings on fluorescent phosphoglycerolipid and sphingolipid analogues in the plasma membrane of live mammalian cells. The STED-FLCS data indicate that biophysical and biochemical parameters such as the affinity for molecular complexes strongly change over space and time within a few seconds. Drug treatment for cholesterol depletion or actin cytoskeleton depolymerization not only results in the already previously observed decreased affinity for molecular interactions but also in a slight reduction of the spatiotemporal heterogeneity. STED-FLCS specifically demonstrates a significant improvement over previous gated STED-FCS experiments and with its improved spatial and temporal resolution is a novel tool for investigating how heterogeneities of the cellular plasma membrane may regulate biofunctionality.

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Language(s): eng - English
 Dates: 2015-08-032015-09
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1021/acs.nanolett.5b02001
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Title: Nano Letters
Source Genre: Journal
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Pages: - Volume / Issue: 15 (9) Sequence Number: - Start / End Page: 5912 - 5918 Identifier: -