Multidimensional Double Refraction Microscopy

Abstract

Multidimensional optical microscopy - extracting 3D location, orientation, and spectral information from nanoscale emitters - is key to modern nanoscience. However, this multidimensional capability requires complex instrumental setups, demanding exceptional stability and specialized expertise in optics. Here, we overcome these barriers by introducing Double Refraction Microscopy (DRM), a passive imaging method providing a straightforward pathway to multidimensional super-resolution imaging. Based on bifocal imaging via a birefringent microscope slide, DRM splits nanoprobe light into a characteristic double image encoding 3D location, orientation, and spectral signature, while requiring zero hardware modifications to standard wide-field microscopes. To showcase its robustness, we implement DRM across three distinct slides of varying materials and thicknesses, calibrating 3D localization with 450-nm nanoparticles and demonstrating high orientation sensitivity using polarization-controlled emitters. Furthermore, we confirm spectral readout using a pinhole array acting as a grid of point emitters back-illuminated at various wavelengths to generate distinct spectral signatures. Finally, we showcase practical utility by performing a two-colour 3D localization measurement within a $10\,μ\text{m}$ gap, utilizing the single optical pathway of a standard epi-fluorescence microscope. Ultimately, DRM provides the missing optical hardware simplicity to complement established computational tools, promising to transfer advanced super-resolution capabilities from specialized facilities to every laboratory bench.