Super resolution imaging of proteins’ organization and conformation in 3D using polarized single molecule localization

Measuring single molecules’ 3D orientations in addition to their 3D spatial localization is still today a challenge, due to the intrinsic coupling of both spatial and orientational parameters in the point spread function (PSF) image formation. We present polarized fluorescence microscopy methods able to report both orientational and spatial information from single molecules with high precision. These methods are applied to STORM and PALM nanoscale imaging of actin filaments organization and membrane proteins’ conformational changes in 3D.