Integrated one- and two-photon scanned oblique plane illumination (SOPi) microscopy for rapid volumetric imaging
Integrated one- and two-photon scanned oblique plane illumination (SOPi) microscopy for rapid volumetric imaging
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DOI：
10.1364/oe.26.013027
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发表时间：
2018-05-14
期刊：
影响因子：
3.8
通讯作者：
Kozorovitskiy, Yevgenia
中科院分区：
文献类型：
--
作者：
Kumar, Manish;Kishore, Sandeep;Kozorovitskiy, Yevgenia
能够在脑深处进行体内快速体积功能和结构成像的多功能空间可达成像系统仍然是神经科学研究的限制因素。为了克服这一障碍，我们提出了集成的单光子和双光子扫描斜平面照明（SOPi，/SOPI）显微镜，它使用一个单一的面向前的显微镜物镜，以提供基于光片扫描的快速体积成像能力在亚细胞分辨率。我们基于平面扫描镜的优化光片架构允许对体积样本进行无失真扫描，从而简化成像体积的准确重建。单光子（1 P）和双光子（2 P）光片显微镜在同一系统中的集成允许在散射介质中的快速体积成像和更高分辨率成像之间进行轻松选择。使用SOPi，我们展示了散射小鼠脑切片内的深，大体积成像能力和快速成像速度高达每秒10卷表达基因编码的荧光蛋白GFP或GCaMP 6s的斑马鱼幼虫。SOPi的灵活性和空间访问使其适用于许多成像应用，并广泛兼容用于驱动或询问神经元结构和活动的正交技术。(C)根据OSA开放获取出版协议的条款，2018年美国光学学会
Versatile, sterically accessible imaging systems capable of in vivo rapid volumetric functional and structural imaging deep in the brain continue to be a limiting factor in neuroscience research. Towards overcoming this obstacle, we present integrated one- and two-photon scanned oblique plane illumination (SOPi, /sopi) microscopy which uses a single front-facing microscope objective to provide light-sheet scanning based rapid volumetric imaging capability at subcellular resolution. Our planar scan-mirror based optimized light-sheet architecture allows for non-distorted scanning of volume samples, simplifying accurate reconstruction of the imaged volume. Integration of both one-photon (1P) and two-photon (2P) light-sheet microscopy in the same system allows for easy selection between rapid volumetric imaging and higher resolution imaging in scattering media. Using SOPi, we demonstrate deep, large volume imaging capability inside scattering mouse brain sections and rapid imaging speeds up to 10 volumes per second in zebrafish larvae expressing genetically encoded fluorescent proteins GFP or GCaMP6s. SOPi's flexibility and steric access makes it adaptable for numerous imaging applications and broadly compatible with orthogonal techniques for actuating or interrogating neuronal structure and activity. (C) 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement