Ultrafast light targeting for high-throughput precise control of neuronal networks.
Ultrafast light targeting for high-throughput precise control of neuronal networks.
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DOI：
10.1038/s41467-023-37416-w
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发表时间：
2023-04-05
影响因子：
16.6
通讯作者：
Emiliani, Valentina
中科院分区：
文献类型：
--
作者：
Faini, Giulia;Tanese, Dimitrii;Molinier, Clement;Telliez, Cecile;Hamdani, Massilia;Blot, Francois;Tourain, Christophe;de Sars, Vincent;Del Bene, Filippo;Forget, Benoit C.;Ronzitti, Emiliano;Emiliani, Valentina
基于全息光靶向方法的双光子、单细胞分辨率光遗传学能够生成精确的时空神经元活动模式，从而实现广泛的实验应用，例如高通量连接映射和探测感知神经代码。然而，目前的全息方法将用于调整不同细胞的相对尖峰时间的分辨率限制为几毫秒，并且根据工作深度，可实现的目标数量限制为100-200。为了克服这些限制并扩展单细胞光遗传学的能力，我们引入了一种基于以kHz速率在全息图之间快速切换时间聚焦光束的超快顺序光靶向（FLiT）光学配置。我们使用FLiT来演示两种照明协议，称为混合和循环照明，并实现体外（小鼠器官型和急性脑切片）和体内（斑马鱼幼虫和小鼠）顺序神经元激活和高通量多细胞照明的亚毫秒控制，同时最大限度地减少光诱导的热上升。这些方法将是重要的实验，需要快速和精确的细胞刺激与定义的时空活动模式和光学控制的大型神经元合奏。目前用于神经元刺激的全息方法在其时间分辨率和目标神经元的数量方面具有限制。在这里，作者展示了一种超快全息光靶向方法，该方法与光遗传学相结合，能够实现亚毫秒级的顺序神经元激活控制和高通量同时多细胞照明。
Two-photon, single-cell resolution optogenetics based on holographic light-targeting approaches enables the generation of precise spatiotemporal neuronal activity patterns and thus a broad range of experimental applications, such as high throughput connectivity mapping and probing neural codes for perception. Yet, current holographic approaches limit the resolution for tuning the relative spiking time of distinct cells to a few milliseconds, and the achievable number of targets to 100-200, depending on the working depth. To overcome these limitations and expand the capabilities of single-cell optogenetics, we introduce an ultra-fast sequential light targeting (FLiT) optical configuration based on the rapid switching of a temporally focused beam between holograms at kHz rates. We used FLiT to demonstrate two illumination protocols, termed hybrid- and cyclic-illumination, and achieve sub-millisecond control of sequential neuronal activation and high throughput multicell illumination in vitro (mouse organotypic and acute brain slices) and in vivo (zebrafish larvae and mice), while minimizing light-induced thermal rise. These approaches will be important for experiments that require rapid and precise cell stimulation with defined spatio-temporal activity patterns and optical control of large neuronal ensembles. Current holographic approaches for neuronal stimulation have limitations in their temporal resolution and the number of targeted neurons. Here, the authors demonstrate an approach for ultra-fast holographic light targeting which, combined with optogenetics, enables sub-millisecond control of sequential neuronal activation and high throughput simultaneous multicell illumination.
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