Light-field microscopy with temporal focusing multiphoton illumination for scanless volumetric bioimaging |
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| Authors: | Feng-Chun Hsu Chun-Yu Lin Yvonne Yuling Hu Yeu-kuang Hwu Ann-Shyn Chiang Shean-Jen Chen |
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| Affiliation: | 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 |
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| 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. |
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