Institute of Neuroscience, Key Laboratory of Brain Cognition and Brain-Inspired Intelligence Technology, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai, China.
University of Chinese Academy of Sciences, Beijing, China.
Nat Methods. 2024 Nov;21(11):2160-2170. doi: 10.1038/s41592-024-02458-5. Epub 2024 Oct 8.
Voltage imaging measures neuronal activity directly and holds promise for understanding information processing within individual neurons and across populations. However, imaging voltage over large neuronal populations has been challenging owing to the simultaneous requirements of high imaging speed and signal-to-noise ratio, large volume coverage and low photobleaching rate. Here, to overcome this challenge, we developed a confocal light-field microscope that surpassed the traditional limits in speed and noise performance by incorporating a speed-enhanced camera, a fast and robust scanning mechanism, laser-speckle-noise elimination and optimized light efficiency. With this method, we achieved simultaneous recording from more than 300 spiking neurons within an 800-µm-diameter and 180-µm-thick volume in the mouse cortex, for more than 20 min. By integrating the spatial and voltage activity profiles, we have mapped three-dimensional neural coordination patterns in awake mouse brains. Our method is robust for routine application in volumetric voltage imaging.
电压成像是一种直接测量神经元活动的方法，有望帮助我们理解单个神经元和神经元群体之间的信息处理过程。然而，由于同时需要高速成像和高信噪比、大体积覆盖和低光漂白率，因此对大量神经元群体进行电压成像一直具有挑战性。为了克服这一挑战，我们开发了一种共聚焦光场显微镜，该显微镜通过结合高速相机、快速稳健的扫描机制、激光散斑噪声消除和优化的光效率，超越了传统速度和噪声性能的限制。通过这种方法，我们在小鼠皮层中直径 800 微米、厚 180 微米的体积内同时记录了超过 300 个放电神经元的活动，持续时间超过 20 分钟。通过整合空间和电压活动谱，我们绘制了清醒小鼠大脑中的三维神经协调模式。我们的方法在体积电压成像的常规应用中具有很强的鲁棒性。
