Eggeling, C. et al. Direct observation of the nanoscale dynamics of membrane lipids in a living cell. Nature 457, 1159-1162

Department of Nanobiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.
Nature (Impact Factor: 42.35). 12/2008; 457(7233):1159-62. DOI: 10.1038/nature07596
Source: PubMed

ABSTRACT Cholesterol-mediated lipid interactions are thought to have a functional role in many membrane-associated processes such as signalling events. Although several experiments indicate their existence, lipid nanodomains ('rafts') remain controversial owing to the lack of suitable detection techniques in living cells. The controversy is reflected in their putative size of 5-200 nm, spanning the range between the extent of a protein complex and the resolution limit of optical microscopy. Here we demonstrate the ability of stimulated emission depletion (STED) far-field fluorescence nanoscopy to detect single diffusing (lipid) molecules in nanosized areas in the plasma membrane of living cells. Tuning of the probed area to spot sizes approximately 70-fold below the diffraction barrier reveals that unlike phosphoglycerolipids, sphingolipids and glycosylphosphatidylinositol-anchored proteins are transiently ( approximately 10-20 ms) trapped in cholesterol-mediated molecular complexes dwelling within <20-nm diameter areas. The non-invasive optical recording of molecular time traces and fluctuation data in tunable nanoscale domains is a powerful new approach to study the dynamics of biomolecules in living cells.

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Available from: Günter Schwarzmann, Aug 19, 2015
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    • "Interestingly, syntaxin-1 molecules are only transiently associated with their nanoclusters and rapidly exchange with freely diffusing syntaxin-1 molecules . Live cell STED imaging also revealed the transient, nanoscale organization of sphingolipids and glycosylphosphatidylinositol (GPI)anchored proteins within a 20 nm cholesterol rich membrane region [53]. These studies support the notion that the PM organization is controlled by dynamic molecular interactions of associated and dissociated states. "
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    Biochimica et Biophysica Acta (BBA) - Molecular Cell Research 11/2014; 1853(4). DOI:10.1016/j.bbamcr.2014.11.010 · 5.30 Impact Factor
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    • "Reducing the focal volume with the STED beam (Figure 4C) revealed the mode of diffusion of phosphoethanolamine, which differed from that of sphingomyelin that becomes temporarily trapped in domains of 520–50 nm for 510 ms (Eggeling et al., 2009). Later work confirmed that shortchained unsaturated lipids diffuse more rapidly than longchain saturated lipids, as predicted by the lipid raft hypothesis (Mueller et al., 2011). "
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    • "Microscopy Research and Technique between antennas and the membrane. Finally, illumination spot size variation as conveniently afforded by STED (Eggeling et al., 2009; Ringemann et al., 2009) is significantly more tedious to implement with antenna probes since it requires the exchange of multiple antennas with different sizes. 2D optical antennas are on the other hand much easier to handle, and they can be made of different sizes on the same substrate. "
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