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- W2914371336 abstract "To date, the only type of membrane in a living cell that is known to phase separate into large (micron-scale) domains is that of the yeast vacuole. In contrast, within plasma membranes, the distribution of lipids and proteins appears to be heterogeneous on short time and length scales. In model membranes containing ternary or quaternary mixtures of lipids, the presence of submicron domains has been inferred by indirect methods such as FRET, neutron diffraction, and NMR. In cases when indirect methods rely on model-dependent assumptions, it is powerful to directly image the membranes. However, direct imaging of submicron domains presents significant experimental challenges. Here, we present two novel cryo electron microscopy methods to directly image domains in submicron vesicles containing ternary and quaternary lipid mixtures. First, we use a probe-free method to image domains that are thicker than the surrounding membrane. To do so, we leverage existing ternary phase diagrams (assessed via fluorescence microscopy) and AFM data to identify lipid types and ratios likely to produce thickness mismatches = 1 nm. Second, we label domains in quaternary mixtures of lipids with an electron-dense protein probe, and we image the distribution of this probe." @default.
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- W2914371336 date "2019-02-01" @default.
- W2914371336 modified "2023-09-30" @default.
- W2914371336 title "Direct Imaging of Membrane Domains in Sub-Micron Lipid Vesicles by Cryo-EM" @default.
- W2914371336 doi "https://doi.org/10.1016/j.bpj.2018.11.463" @default.
- W2914371336 hasPublicationYear "2019" @default.
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