Fast, cell-resolution, contiguous-wide two-photon imaging to reveal functional network architectures across multi-modal cortical areas
Fast, cell-resolution, contiguous-wide two-photon imaging to reveal functional network architectures across multi-modal cortical areas
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DOI：
10.1016/j.neuron.2021.03.032
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发表时间：
2021-06-02
期刊：
影响因子：
16.2
通讯作者：
Murayama, Masanori
中科院分区：
文献类型：
--
作者：
Ota, Keisuke;Oisi, Yasuhiro;Murayama, Masanori
具有单细胞分辨率、高信噪比和无光学像差的快速和宽视场成像有可能激发生物学研究的新途径。然而，这种成像是具有挑战性的，因为这些参数之间不可避免的权衡。在这里，我们克服了这些权衡相结合的共振扫描系统，一个大的物镜与低放大倍率和高数值孔径，高灵敏度的大孔径光电探测器。其结果是一种几乎无像差、快速扫描的高光学不变双光子显微镜（FASHIO-2PM），能够从一个由16，000个神经元组成的大型网络以7.5 Hz的频率从9 mm（2）连续图像平面进行钙成像，包括清醒小鼠大脑皮层的10多个感觉运动和高阶区域。基于单细胞活动的网络分析表明，大脑表现出小世界而不是无标度行为。预计FASHIO-2PM将通过同时监测宏观活动及其组成元素来进行生物动力学研究。
Fast and wide field-of-view imaging with single-cell resolution, high signal-to-noise ratio, and no optical aberrations have the potential to inspire new avenues of investigations in biology. However, such imaging is challenging because of the inevitable tradeoffs among these parameters. Here, we overcome these tradeoffs by combining a resonant scanning system, a large objective with low magnification and high numerical aperture, and highly sensitive large-aperture photodetectors. The result is a practically aberration-free, fast-scanning high optical invariant two-photon microscopy (FASHIO-2PM) that enables calcium imaging from a large network composed of similar to 16,000 neurons at 7.5 Hz from a 9 mm(2) contiguous image plane, including more than 10 sensory-motor and higher-order areas of the cerebral cortex in awake mice. Network analysis based on single-cell activities revealed that the brain exhibits small-world rather than scale-free behavior. The FASHIO-2PM is expected to enable studies on biological dynamics by simultaneously monitoring macroscopic activities and their compositional elements.