Kilohertz frame-rate two-photon tomography
Kilohertz frame-rate two-photon tomography
复制标题
DOI：
10.1038/s41592-019-0493-9
复制
发表时间：
2019-08-01
期刊：
影响因子：
48
通讯作者：
Podgorski, Kaspar
中科院分区：
文献类型：
--
作者：
Kazemipour, Abbas;Novak, Ondrej;Podgorski, Kaspar
点扫描双光子显微镜可以在哺乳动物大脑等散射样本中进行高分辨率成像，但体素的连续采集从根本上限制了其速度。我们开发了一种双光子成像技术，可以以多个角度扫描焦平面上的激发线，并通过计算恢复高分辨率图像，从而在结构化样本中获得超过 10 亿赫兹的体素率。使用静态图像作为记录神经活动的先验，我们对小鼠数百个树突棘的视觉诱发和自发谷氨酸释放进行成像，深度超过 250 μm，帧速率超过 1 kHz。麻醉小鼠中的树突状谷氨酸瞬态在跨越数十微米的空间连续域内同步，频率范围为 1-100 Hz。我们展示了乙酰胆碱和电压指示器的毫秒分辨率记录、密集标记皮层中的三维单粒子跟踪和成像。我们的方法超越了荧光寿命对光栅扫描成像速度的限制。
Point-scanning two-photon microscopy enables high-resolution imaging within scattering specimens such as the mammalian brain, but sequential acquisition of voxels fundamentally limits its speed. We developed a two-photon imaging technique that scans lines of excitation across a focal plane at multiple angles and computationally recovers high-resolution images, attaining voxel rates of over 1 billion Hz in structured samples. Using a static image as a prior for recording neural activity, we imaged visually evoked and spontaneous glutamate release across hundreds of dendritic spines in mice at depths over 250 mu m and frame rates over 1 kHz. Dendritic glutamate transients in anesthetized mice are synchronized within spatially contiguous domains spanning tens of micrometers at frequencies ranging from 1-100 Hz. We demonstrate millisecond-resolved recordings of acetylcholine and voltage indicators, three-dimensional single-particle tracking and imaging in densely labeled cortex. Our method surpasses limits on the speed of raster-scanned imaging imposed by fluorescence lifetime.