High-speed in vivo calcium imaging reveals neuronal network activity with near-millisecond precision
High-speed in vivo calcium imaging reveals neuronal network activity with near-millisecond precision
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DOI：
10.1038/nmeth.1453
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发表时间：
2010-05-01
期刊：
影响因子：
48
通讯作者：
Helmchen, Fritjof
中科院分区：
文献类型：
--
作者：
Grewe, Benjamin F.;Langer, Dominik;Helmchen, Fritjof
神经元群体的双光子钙成像能够光学记录活体动物的尖峰活动，但标准激光扫描仪太慢，无法准确确定尖峰时间。在这里，我们报告在小鼠新皮层的体内成像大大提高了时间分辨率，使用随机访问扫描声光偏转器。我们在180-490 Hz采样率下获得了34-91个2/3层神经元的荧光测量。我们检测单个动作电位诱发的钙瞬变，信噪比为2-5，并以近毫秒的精度和5-15 ms的置信区间确定尖峰时间。一个自动化的“剥离”算法，使复杂的尖峰列车重建荧光痕迹高达20-30赫兹的频率，揭示时空试验到试验的变化，在桶皮层和视觉皮层的感觉反应。通过在快速时间尺度上揭示神经元群体中的尖峰序列，高速钙成像将促进脑微电路中信息处理的光学研究。
Two-photon calcium imaging of neuronal populations enables optical recording of spiking activity in living animals, but standard laser scanners are too slow to accurately determine spike times. Here we report in vivo imaging in mouse neocortex with greatly improved temporal resolution using random-access scanning with acousto-optic deflectors. We obtained fluorescence measurements from 34-91 layer 2/3 neurons at a 180-490 Hz sampling rate. We detected single action potential-evoked calcium transients with signal-to-noise ratios of 2-5 and determined spike times with near-millisecond precision and 5-15 ms confidence intervals. An automated 'peeling' algorithm enabled reconstruction of complex spike trains from fluorescence traces up to 20-30 Hz frequency, uncovering spatiotemporal trial-to-trial variability of sensory responses in barrel cortex and visual cortex. By revealing spike sequences in neuronal populations on a fast time scale, high-speed calcium imaging will facilitate optical studies of information processing in brain microcircuits.