IRFP: Scanless Two-Photon Voltage Imaging of Live Neuron Activity With Holographic Wavefront Shaping

国际研究奖学金项目使美国科学家和工程师能够在国外进行9至24个月的研究。该计划的奖励为联合研究提供了机会，并利用独特或互补的设施、专业知识和国外的实验条件。该奖项将支持Amanda J. Foust博士与Valentina Emiliani博士在法国巴黎笛卡尔大学开展为期24个月的研究。电压成像有可能彻底改变我们对神经元细胞和电路如何通过模拟和数字电信号在活体大脑中混合处理和存储信息的理解。然而，低信噪比和高背景妨碍了足够的空间和时间分辨率来跟踪体内细胞和亚细胞长度尺度上的电活动。瓦伦蒂娜·埃米利亚尼博士的实验室正在开发的梯度相位对比（GPC）和时间聚焦（TF）等光雕刻技术可以通过对特定细胞和亚结构进行定向荧光激发来提高电压成像技术的空间分辨率。为此，我们正在开发、调整和优化波前整形技术，利用电压敏感染料和荧光蛋白对神经元轴突和树突中的电活动传播进行成像。空间定向电压传感的波前整形技术的发展将为研究感觉、感知、认知和行为的电基础提供有力的工具。此外，这些技术可用于确定癫痫和多发性硬化症等神经系统疾病和紊乱中发生的基本电路改变，并为开发更有效的治疗策略提供信息。

The International Research Fellowship Program enables U.S. scientists and engineers to conduct nine to twenty-four months of research abroad. The program's awards provide opportunities for joint research, and the use of unique or complementary facilities, expertise and experimental conditions abroad. This award will support a twenty-four month research fellowship by Dr. Amanda J. Foust to work with Dr. Valentina Emiliani at Université Paris Descartes, Paris, France. Voltage imaging has the potential to revolutionize our understanding of how neuronal cells and circuits process and store information through a mixture of analog and digital electrical signaling in the living brain. However, low signal-to-noise ratios and high background have prevented spatial and temporal resolution adequate for tracking electrical activity on cellular and subcellular length scales in-vivo. Light sculpturing techniques such as Gradient Phase Contrast (GPC) and temporal focusing (TF) under development in Dr. Valentina Emiliani's laboratory can improve the spatial resolution of voltage imaging techniques by enabling targeted fluorescence excitation of specified cells and substructures. To this end, we are developing, adapting, and optimizing wavefront shaping technology for imaging the propagation of electrical activity in the axons and dendrites of neurons with voltage-sensitive dyes and fluorescent proteins. Development of wavefront shaping technology for spatially targeted voltage sensing will provide a powerful tool for investigating the electrical underpinnings of sensation, perception, cognition, and behavior. Moreover, these techniques can be used to determine what fundamental circuit alterations occur with neurological diseases and disorders such as epilepsy and multiple sclerosis, and inform development of more effective therapeutic strategies.