Figure 1.

Polarized localization microscopy combines the techniques of polarized TIRFM and SMLM. By controlling the linear polarization of incident excitation light, the electric field (green arrows) of the evanescent wave for fluorescence excitation can be either (A) vertical with p-polarized light or (B) horizontal with s-polarized light. This results in differential excitation of rotationally confined fluorophores dependent on the local membrane orientation. (C) Imaging and localizing individual blinking fluorophores in separate frames enables the reconstruction of superresolution images with embedded information on membrane orientations. (D) The probability of localizing a DiI depends on the direction of the normal vector to the membrane (n) relative to the coverslip, as described by θ and φ, as well as and the angle of the DiI dipole moment (μ) within the membrane, as described by β and ψ. To see this figure in color, go online.