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Short summary of the pixOL technique @[osf](zfdbt) @[osf](w9ecr) Demonstration of SM detection and position orientation estimation using the pixOL microscope. (Top) Polarized images (red: x-polarized and blue: y-polarized) of Nile red are compared to (bottom) reconstructed images of the pixOL DSF using 3D orientations and 3D positions estimated by a bespoke regularized maximum likelihood estimator. Individual Nile red molecules are shown binding transiently to lipids (DPPC with cholesterol) within a spherical bilayer. Colorbar: photons/pixel. Scale bar: 1 μm. @[osf](dr92x) 3D view of 6D single-molecule orientation localization microscopy (SMOLM) imaging of Nile red within spherical supported lipid bilayers consisting of (top) DPPC plus cholesterol (chol) and (bottom) DPPC-only. The position and orientation of each line depicts an emitter’s 3D position and 3D orientation. Colors represent polar orientation θ. @[osf](6a54z) x-y cross-sections of 6D SMOLM imaging of Nile red within spherical supported lipid bilayers consisting of (top) DPPC plus cholesterol (chol) and (bottom) DPPC only at various h-slices. The position and orientation of each line depicts an emitter’s lateral position and 3D orientation. Colors represent azimuthal orientation ϕ.
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