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A phase mask design that we term a “bisected pupil”(BSP) provides several advantages for single-molecule optical imaging. When using the BSP with a dual-polarization optical Fourier processing system, both the position and dipole orientation of individual fluorescent molecules may be measured from a single camera image. In the context of single-molecule super-resolution microscopy, this technique permits one to diagnose, and subsequently to remove imaging artifacts resulting from orientation-induced localization errors. If the molecules labeling a structure are rotationally mobile, thus mitigating dipole orientation errors, this technique enables super-resolution imaging in three dimensions. We present simulations and experimental verification. VC 2014 AIP Publishing LLC.[http://dx. doi. org/10.1063/1.4876440]

The field of fluorescence microscopy has made remarkable progress in recent years, benefitting from …