Fast two-photon imaging of subcellular voltage dynamics in neuronal tissue with genetically encoded indicators
Fast two-photon imaging of subcellular voltage dynamics in neuronal tissue with genetically encoded indicators
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DOI：
10.7554/elife.25690
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发表时间：
2017-07-27
期刊：
影响因子：
7.7
通讯作者：
St-Pierre, Francois
中科院分区：
文献类型：
--
作者：
Chamberland, Simon;Yang, Helen H.;St-Pierre, Francois
监测大脑深处特定神经元的电压动态对于揭示神经元回路的功能至关重要，但由于现有工具的性能有限，因此具有挑战性。特别是，虽然基因编码的电压指示器已经显示出对电压瞬变的光学检测的希望，但许多指示器在双光子照明下成像时表现出低灵敏度。因此，以前的研究在单个试验中缺乏对单个神经元电压动态的可视化。在这里，我们报告了ASAP2s，一种新型电压指示器，具有更高的灵敏度。通过使用随机接入多光子显微镜对ASAP2s进行成像，我们证明了在有机型切片培养中单次试验检测动作电位的可靠性。我们还发现ASAP2s能够在器官型切片培养和果蝇中实现梯度电位的双光子成像。这些结果表明，结合ASAP2s和快速双光子成像方法可以实现亚细胞空间分辨率和毫秒时间尺度精度的神经电活动检测。
Monitoring voltage dynamics in defined neurons deep in the brain is critical for unraveling the function of neuronal circuits but is challenging due to the limited performance of existing tools. In particular, while genetically encoded voltage indicators have shown promise for optical detection of voltage transients, many indicators exhibit low sensitivity when imaged under two-photon illumination. Previous studies thus fell short of visualizing voltage dynamics in individual neurons in single trials. Here, we report ASAP2s, a novel voltage indicator with improved sensitivity. By imaging ASAP2s using random-access multi-photon microscopy, we demonstrate robust single-trial detection of action potentials in organotypic slice cultures. We also show that ASAP2s enables two-photon imaging of graded potentials in organotypic slice cultures and in Drosophila. These results demonstrate that the combination of ASAP2s and fast two-photon imaging methods enables detection of neural electrical activity with subcellular spatial resolution and millisecond-timescale precision.