Axial standing-wave illumination frequency-domain imaging (SWIF)

Abstract

Despite their tremendous contribution to biomedical research and diagnosis, conventional spatial sampling techniques such as wide-field, point scanning or selective plane illumination microscopy face inherent limiting trade-offs between spatial resolution, field-of-view, phototoxicity and recording speed. Several of these trade-offs are the result of spatial sampling with diffracting beams. Here, we introduce a new strategy for fluorescence imaging, SWIF, which instead encodes the axial profile of a sample in the Fourier domain. We demonstrate how this can be achieved with propagation-invariant illumination patterns that extend over several millimeters and robustly propagate through layers of varying refractive index. This enabled us to image a lateral field-of-view of 0.8 mm x 1.5 mm with an axial resolution of 2.4 µm – greatly exceeding the lateral field-of-view of conventional illumination techniques (~100 µm) at comparable resolution. Thus, SWIF allowed us to surpass the limitations of diffracting illumination beams and untangle lateral field-of-view from resolution.

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