Hybrid light-sheet and light-field microscope for high resolution and large volume neuroimaging
Hybrid light-sheet and light-field microscope for high resolution and large volume neuroimaging
复制标题
DOI：
10.1364/boe.10.006595
复制
发表时间：
2019-12-01
影响因子：
3.4
通讯作者：
Gong, Yiyang
中科院分区：
文献类型：
--
作者：
Wang, Depeng;Xu, Stephen;Gong, Yiyang
大规模同时记录神经活动的快速模式仍然具有挑战性。扫描光束光片显微镜 (LSM) 和宽视场光场显微镜 (WFLFM) 等体积成像方式由于其复杂的校准程序、低空间分辨率或高光漂白而未能达到目标。在这里，我们展示了一种混合光片光场显微镜（LSLFM）模式，该模式可通过简化成像平面和激发平面的对准来产生高空间分辨率。这种新模式将光片照明的选择性激发与体积光场成像结合起来。这种方式克服了扫描光束 LSM 和 WFLFM 实现的当前限制。与LSM相比，LSFLM以比LSM低50倍的帧速率捕获体积数据，并且不需要动态校准。与 WFLFM 相比，LSFLM 的空间分辨率有所提高，荧光珠成像时的对比度提高了 10 倍，对表达基因编码钙传感器的活体斑马鱼进行成像时，神经活动检测的信噪比提高了 3.2 倍。 (C) 2019 年美国光学学会根据 OSA 开放获取出版协议条款
Large scale simultaneous recording of fast patterns of neural activity remains challenging. Volumetric imaging modalities such as scanning-beam light-sheet microscopy (LSM) and wide-field light-field microscopy (WFLFM) fall short of the goal due to their complex calibration procedure, low spatial resolution, or high-photobleaching. Here, we demonstrate a hybrid light-sheet light-field microscopy (LSLFM) modality that yields high spatial resolution with simplified alignment of the imaging plane and the excitation plane. This new modality combines the selective excitation of light-sheet illumination with volumetric light-field imaging. This modality overcomes the current limitations of the scanning-beam LSM and WFLFM implementations. Compared with LSM, LSLFM captures volumetric data at a frame rate 50 x lower than the rate of LSM and requires no dynamic calibration. Compared with WFLFM, LSLFM produces moderate improvements in spatial resolutions, 10 times improvement in the contrast when imaging fluorescent beads, and 3.2 x the signal-to-noise ratio in the detection of neural activity when imaging live zebrafish expressing a genetically encoded calcium sensor. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement