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Super-resolution mapping of the 3D orientation of fluorophores reveals thealignment of biological structures where the fluorophores are tightly attached,and thus plays a vital role in studying the organization and dynamics ofbio-complexes. However, current super-resolution imaging techniques are eitherlimited to 2D orientation mapping or suffer from slow speed and the requirementof special labels in 3D orientation mapping. Here, we propose a novel polarizedvirtual spatial-frequency-shift effect to overcome these restrictions toachieve a universal 3D orientation super-resolution mapping capability. Todemonstrate the mechanism, we simulate the imaging process and reconstruct thespatial-angular information for sparsely distributed dipoles with random 3Dorientations and microfilament-like structures decorated with fluorophoresoriented parallel to them. The 3D orientation distribution can be recoveredwith a doubled spatial resolution and an average angular precision of up to2.39 degrees. The performance of the approach with noise has also been analyzedconsidering real implementation.

3D orientation super-resolution spatial-frequency-shift microscopy