Light-field microscopy with temporal focusing multiphoton illumination for scanless volumetric bioimaging

Feng-Chun Hsu; Chun-Yu Lin; Yvonne Yuling Hu; Yeu-Kuang Hwu; Ann-Shyn Chiang; Shean-Jen Chen

doi:10.1364/BOE.473807

Light-field microscopy with temporal focusing multiphoton illumination for scanless volumetric bioimaging

Biomed Opt Express. 2022 Nov 23;13(12):6610-6620. doi: 10.1364/BOE.473807. eCollection 2022 Dec 1.

Authors

Feng-Chun Hsu¹, Chun-Yu Lin¹, Yvonne Yuling Hu², Yeu-Kuang Hwu³, Ann-Shyn Chiang⁴, Shean-Jen Chen^{1

5}

Affiliations

¹ College of Photonics, National Yang Ming Chiao Tung University, Tainan 112, Taiwan.
² Department of Photonics, National Cheng Kung University, Tainan 701, Taiwan.
³ Institute of Physics, Academia Sinica, Taipei 115, Taiwan.
⁴ Brain Research Center, National Tsing Hua University, Hsinchu 300, Taiwan.
⁵ Taiwan Instrument Research Institute, National Applied Research Laboratories, Hsinchu 300, Taiwan.

Abstract

A temporal focusing multiphoton illumination (TFMI) method is proposed for achieving selective volume illumination (SVI) (i.e., illuminating only the volume of interest) in light-field microscopy (LFM). The proposed method minimizes the background noise of the LFM images and enhances the contrast, and thus improves the imaging quality. Three-dimensional (3D) volumetric imaging is achieved by reconstructing the LFM images using a phase-space deconvolution algorithm. The experimental results obtained using 100-nm fluorescent beads show that the proposed TFMI-LFM system achieves lateral and axial resolutions of 1.2 µm and 1.1 µm, respectively, at the focal plane. Furthermore, the TFMI-LFM system enables 3D images of the single lobe of the drosophila mushroom body with GFP biomarker (OK-107) to be reconstructed in a one-snapshot record.