Improved and fast biotissue imaging by temporal focusing widefield multiphoton microscopy with spatially modulated illumination

Chia-Yuan Chang; Chun-Yu Lin; Vladimir A. Hovhannisyan; Shean-Jen Chen

doi:10.1117/12.2526908

31 July 2019 Improved and fast biotissue imaging by temporal focusing widefield multiphoton microscopy with spatially modulated illumination

Chia-Yuan Chang, Chun-Yu Lin, Vladimir A. Hovhannisyan, Shean-Jen Chen

Author Affiliations +

Proceedings Volume 11076, Advances in Microscopic Imaging II; 110760G (2019) https://doi.org/10.1117/12.2526908
Event: European Conferences on Biomedical Optics, 2019, Munich, Germany

Abstract

A developed temporal focusing-based multiphoton microscope (TFMPM) has a digital micromirror device (DMD) which is adopted not only as a blazed grating for light spatial dispersion but also for patterned illumination simultaneously. Herein, the TFMPM has been extended to implement spatially modulated illumination at structured frequency and orientation to increase the beam coverage at the back-focal aperture of the objective lens. The axial excitation confinement (AEC) of TFMPM can be condensed from 3.0 μm to 1.5 μm for a 50% improvement. Furthermore, a multiline scanning mechanism based on the DMD can push its AEC nearly equivalent to line scanningbased TFMPM to 1.5 μm from optimal 3.0 μm of the conventional TFMPM. By using the TFMPM with structured illumination and multiline scanning, reconstructed biotissue images according to the condensed AEC structured illumination are shown obviously superior in contrast and better scattering suppression.

Citation Download Citation

Chia-Yuan Chang, Chun-Yu Lin, Vladimir A. Hovhannisyan, and Shean-Jen Chen "Improved and fast biotissue imaging by temporal focusing widefield multiphoton microscopy with spatially modulated illumination", Proc. SPIE 11076, Advances in Microscopic Imaging II, 110760G (31 July 2019); https://doi.org/10.1117/12.2526908

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