Improvement of axial excitation confinement in temporal focusing-based multiphoton microscopy via spatially modulated illumination

Chia-Yuan Chang; Shean-Jen Chen

doi:10.1117/12.2251183

20 February 2017 Improvement of axial excitation confinement in temporal focusing-based multiphoton microscopy via spatially modulated illumination

Chia-Yuan Chang, Shean-Jen Chen

Author Affiliations +

Proceedings Volume 10117, Emerging Digital Micromirror Device Based Systems and Applications IX; 101170F (2017) https://doi.org/10.1117/12.2251183
Event: SPIE OPTO, 2017, San Francisco, California, United States

Abstract

Conventional temporal focusing-based multiphoton excitation microscopy (TFMPEM) can offer widefield optical sectioning with an axial excitation confinement (AEC) of a few microns. Herein, a developed TFMPEM with a digital micromirror device (DMD), acting as the blazed grating for light spatial dispersion and simultaneous patterned illumination, has been extended to implement spatially modulated illumination at structured frequency and orientation. By implementing the spatially modulated illumination, the beam coverage at the back-focal aperture of the objective lens can be increased. As a result, the AEC can be condensed from 3.0 μm to 1.5 μm in full width at half maximum for a 2-fold enhancement. Furthermore, by using HiLo microscopy with two structured illuminations at the same spatial frequency but different orientation, biotissue images according to the structured illumination with condensed AEC is obviously superior in contrast and scattering suppression.

Conference Presentation

Citation Download Citation

Chia-Yuan Chang and Shean-Jen Chen "Improvement of axial excitation confinement in temporal focusing-based multiphoton microscopy via spatially modulated illumination", Proc. SPIE 10117, Emerging Digital Micromirror Device Based Systems and Applications IX, 101170F (20 February 2017); https://doi.org/10.1117/12.2251183

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