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Fluorescence polarization microscopy images both the intensity andorientation of fluorescent dipoles, which plays a vital role in studying themolecular structure and dynamics of bio-complex. However, it is difficult toresolve the dipole assemblies on the subcellular structure and their dynamicsin living cells with super-resolution. Here we report polarized structuredillumination microscopy (pSIM), which decouples the entangled spatial andangular structured illumination through interpreting the dipoles inspatio-angular hyperspace. We demonstrate its application on a series ofbiological filamentous systems such as cytoskeleton networks and lambda-DNA,and report the dynamics of short actin sliding through myosin-coated surface.Further, pSIM reveals "side-by-side" organization of the actin ring structurein the membrane-associated periodic skeleton in hippocampal neurons. It alsoimages the dipole dynamics of green fluorescent proteins labeled to themicrotubules in live U2OS cells. pSIM can be applied directly to a largevariety of commercial or home-built SIM systems.

Super-resolution Imaging of the Fluorescent Dipole Assembly with Polarized Structured Illumination Microscopy