Large-scale calcium imaging with a head-mount axial scanning 3D fluorescence microscope
Large-scale calcium imaging with a head-mount axial scanning 3D fluorescence microscope
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使用头戴式轴向扫描 3D 荧光显微镜进行大规模钙成像
DOI：
10.1101/2021.01.20.427512
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Kobayakawa Reiko
中科院分区：
文献类型：
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作者：
Hayashi Yuichiro;Kobayakawa Ko;Kobayakawa Reiko
微型荧光显微镜对于破译各种大脑功能背后的神经代码变得越来越重要。通过梯度折射率（GRIN）透镜，这些设备能够记录大脑深部结构中的神经元活动。然而，为了最小化对脑组织和局部回路的任何损伤，GRIN透镜的直径应该为0.5-1 mm，从而导致小的视场。考虑到大脑中神经回路的三维（3D）结构，体积成像能力可能会增加通过透镜成像的神经元数量。为了观察三维钙动力学，我们开发了一种小型化的显微镜与电可调透镜和一种新的CNMF为基础的神经信号提取算法的宽场三维成像数据。通过结合硬件和软件，大约1000个神经元从自由行为的小鼠的皮质成像。与最先进的2D成像技术相比，所提出的3D方法可以成像1.7-2.6倍的细胞，并且细胞信号的分离度更高。
Miniaturized fluorescence microscopes are becoming more important for deciphering the neural codes underlying various brain functions. With gradient index (GRIN) lenses, these devices enable recording neuronal activity in deep brain structures. However, to minimize any damage to brain tissues and local circuits, the diameter of the GRIN lens should be 0.5–1 mm, resulting in a small field of view. Volumetric imaging capability might increase the number of neurons imaged through the lenses considering the three-dimensional (3D) structure of neural circuits in the brain. To observe 3D calcium dynamics, we developed a miniaturized microscope with an electrically tunable lens and a novel CNMF-based neural signal extraction algorithm for wide-field 3D imaging data. By combining the hardware and software, approximately 1000 neurons were imaged from the cortices of freely behaving mice. Compared with the state-of-the-art 2D imaging technique, the proposed 3D method imaged 1.7–2.6 times more cells with a higher separation of cellular signals.