Polarized microscopy for single molecule’s orientation and localization imaging

Measuring single molecules’ 3D orientations simultaneously with their 3D spatial localization is currently a topic of intense efforts in optical design. We have developed different methods based on polarized imaging, that are capable to report both 3D spatial localization and 3D orientation (including wobbling) from single molecules. We present how the design of such schemes can be adapted to account from experimental constraints versus accuracy and precision of the estimation of both orientation and localization parameters. We illustrate such performances on actin structural STORM imaging in complex and dense meshworks in fixed cells.