Widefield imaging of rapid pan-cortical voltage dynamics with an indicator evolved for one-photon microscopy.
Widefield imaging of rapid pan-cortical voltage dynamics with an indicator evolved for one-photon microscopy.
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DOI：
10.1038/s41467-023-41975-3
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发表时间：
2023-10-12
影响因子：
16.6
通讯作者：
St-Pierre, Francois
中科院分区：
文献类型：
--
作者：
Lu, Xiaoyu;Wang, Yunmiao;Liu, Zhuohe;Gou, Yueyang;Jaeger, Dieter;St-Pierre, Francois
使用基因编码电压指示器 (GEVI) 的宽场成像是了解大型皮层网络在行为神经编码中的作用的一种很有前景的方法。然而，当前 GEVI 的性能有限限制了它们在快速神经元电压动态的单次试验成像中的部署。在这里，我们开发了一个高通量平台来筛选 GEVI，该平台在宽场单光子照明下将快速动力学与高亮度、灵敏度和光稳定性结合起来。经过多轮定向进化产生了 JEDI-1P，这是一种在所有指标上都具有增强性能的绿色荧光指示剂。接下来，我们优化了新生儿脑室内递送方法，以在小鼠中实现经济有效且广泛的 JEDI-1P 表达。我们还开发了一种方法，可以有效地纠正血流动力学和运动伪影的光学测量结果。最后，我们实现了稳定的全脑电压成像，并在单次试验中成功跟踪了清醒小鼠的伽马频率胡须和视觉刺激，为研究高频信号在大脑计算中的作用打开了大门。基因编码的电压指示器需要快速且大的信号，以实现快速神经元活动的宽场成像。在这里，作者开发了 JEDI-1P 指示器，并在单次试验中展示了清醒小鼠的全皮质电压成像和伽马频率跟踪。
Widefield imaging with genetically encoded voltage indicators (GEVIs) is a promising approach for understanding the role of large cortical networks in the neural coding of behavior. However, the limited performance of current GEVIs restricts their deployment for single-trial imaging of rapid neuronal voltage dynamics. Here, we developed a high-throughput platform to screen for GEVIs that combine fast kinetics with high brightness, sensitivity, and photostability under widefield one-photon illumination. Rounds of directed evolution produced JEDI-1P, a green-emitting fluorescent indicator with enhanced performance across all metrics. Next, we optimized a neonatal intracerebroventricular delivery method to achieve cost-effective and wide-spread JEDI-1P expression in mice. We also developed an approach to correct optical measurements from hemodynamic and motion artifacts effectively. Finally, we achieved stable brain-wide voltage imaging and successfully tracked gamma-frequency whisker and visual stimulations in awake mice in single trials, opening the door to investigating the role of high-frequency signals in brain computations. Genetically encoded voltage indicators need fast and large signals to allow widefield imaging of rapid neuronal activity. Here, the authors develop the indicator JEDI-1P and demonstrate pan-cortical voltage imaging and gamma-frequency tracking in awake mice in single trials.
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影响因子：
2.1
作者：
Ghasemi A;Zahediasl S
通讯作者：
Zahediasl S
影响因子：
25
作者：
Chan KY;Jang MJ;Yoo BB;Greenbaum A;Ravi N;Wu WL;Sánchez-Guardado L;Lois C;Mazmanian SK;Deverman BE;Gradinaru V
通讯作者：
Gradinaru V
影响因子：
3.4
作者：
Brinks, Daan;Klein, Aaron J.;Cohen, Adam E.
通讯作者：
Cohen, Adam E.
影响因子：
16.2
作者：
Ferezou, Isabelle;Haiss, Florent;Petersen, Carl C. H.
通讯作者：
Petersen, Carl C. H.
DOI：
10.1126/science.aab0810
发表时间：
2015-12-11
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gong Y;Huang C;Li JZ;Grewe BF;Zhang Y;Eismann S;Schnitzer MJ
通讯作者：
Schnitzer MJ