Light Field Multiphoton Microscopy with Temporal Focusing-Based Volume Selective Excitation

Feng Chun Hsu, Yong Da Sie, Chun Yu Lin, Yvonne Yuling Hu, Shean Jen Chen*


研究成果: Conference contribution同行評審


Light field fluorescence microscopy (LFM) can provide three-dimensional (3D) images in one snapshot, but essentially lighting up the entirety of the sample, even though only a part of the sample is meaningfully captured in the reconstruction. Thus, entire illumination introduces extraneous background noise, degrading the contrast and accuracy of the final reconstructed images. In this paper, temporal focusing-based multiphoton illumination (TFMI) has the advantage of widefield multiphoton excitation with volume selective excitation. We implement the TFMI to LFM, illuminating only the volume of interest, thus significantly reducing the background. Furthermore, offering higher penetration depth in scattering tissue via multiphoton. In addition, the volume range can be varied by modulating the size of the Fourier-plane aperture of objective lens. 100 nm fluorescence beads are used to examine the lateral and axial resolution after phase space deconvolution from light field image, the experimental results show that the lateral resolution is around 1.2 µm and axial resolution is around 1.6 µm close to the focal plane. Furthermore, the mushroom body of drosophila brain which carried a genetic fluorescent marker GFP (OK-107) are used to demonstrate volumetric bioimaging capability.

主出版物標題Unconventional Optical Imaging III
編輯Marc P. Georges, Gabriel Popescu, Nicolas Verrier
出版狀態Published - 2022
事件Unconventional Optical Imaging III 2022 - Virtual, Online
持續時間: 9 5月 202220 5月 2022


名字Proceedings of SPIE - The International Society for Optical Engineering


ConferenceUnconventional Optical Imaging III 2022
城市Virtual, Online


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