Programmable Aperture Light-Field Microscopy

doi:10.1002/lpor.202300217

2023-04-10

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DOI：10.1002/lpor.202300217

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摘要

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A computational three-dimensional (3D) microscopy technique, termed programmable aperture light-field microscopy (PALFM), for motion-free, high-resolution volumetric imaging of fluorescent or self-luminous samples is proposed. The well-known Fourier slice theorem is extended to incoherent tomographic imaging, which states that a detected image under an ideal aperture corresponds to a central slice in the 3D object spectrum, so the spectrum coverage can be accomplished based on the motion-free aperture modulation. When further considering frequency extension and coverage, a hybrid aperture modulation scheme is designed consisting of non-centrosymmetric circular and annular apertures for high-efficiency, non-ambiguous depth discrimination. A PALFM system with an easy-to-build programmable aperture module attached to an off-the-shelf inverted fluorescence microscope is constructed, where annular apertures can manipulate Bessel-like beams for spectrum modulation. Experimental results on near-diffraction-limited imaging of a resolution target across a large depth range and high-resolution, multi-color 3D imaging of a mouse kidney section verify the validity and effectiveness of PALFM. High-speed, long-term time-lapse volumetric imaging of HeLa cells in vitro further demonstrates that PALFM is a promising tomographic imaging tool for studying dynamic cellular processes and events without requiring complicated sample rotation or beam scanning.

Keyword:

aperture modulation

incoherent Fourier slice theorem

light-field microscopy

tomography

volumetric imaging