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Holography-based structured light illumination for temporal focusing microscopy

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Abstract

In this Letter, we present a holography-based structured light illumination (SLI) method to enhance the resolution of widefield temporal focusing microscopy (TFM). In the system, a digital micromirror device is employed to simultaneously disperse the incoming femtosecond laser to induce temporal focusing at the focal plane and generate designed structured patterns via a Lee hologram. As the generated structured patterns do not contain the zeroth order beam, it improves the contrast and modulation frequency. Mathematical models have been derived to calculate the electric fields at the focal plane and to explain the effects of improved optical cross-sectioning capability. Imaging experiments have been devised and performed on fluorescent beads and mouse kidney sections; the results demonstrate enhanced axial confinement and improved suppression of out-of-focus fluorescence. The new SLI method realizes high-resolution TFM and can be readily applied to other microscopy platforms for biophotonics applications.

© 2021 Optical Society of America

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Supplementary Material (1)

NameDescription
Visualization 1       3D imaging of a mouse kidney sample: (left) widefield TFM images; (middle) conventional SLI-TFM; and (right) holography-based SLI-TFM, over a volume 80 × 80 × 18 µm3 at a speed of 10 frames per second. The excitation laser power is 2 mW.

Data Availability

Data underlying the results presented in this paper are not publicly available at this time but may be obtained from the authors upon reasonable request.

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