Collaborative Research: NCS-FO: Modified two-photon microscope with high-speed electrowetting array for imaging voltage transients in cerebellar molecular layer interneurons

合作研究：NCS-FO：带有高速电润湿阵列的改良双光子显微镜，用于对小脑分子层中间神经元的电压瞬变进行成像

• 批准号: 2319406
• 负责人: Emily Gibson
• 金额: $ 52.5万
• 依托单位: University of Colorado at Denver-Downtown Campus
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2319406&HistoricalAwards=false
• 关键词: Collaborative; Research; NCS; FO; Modified

The research project aims to develop a revolutionary microscope for studying the brain's neural activity. By combining expertise from engineering, physics, and neuroscience, the multidisciplinary team is working on a high-risk, high-payoff endeavor to overcome the limitations of existing imaging techniques. The investigators are focusing on genetically encoded voltage indicators (GEVIs) to capture fast neuronal firing and subthreshold voltage changes. The innovative approach combines two-photon microscopy with electrowetting adaptive optical microprism technology, enabling selective excitation and imaging of groups of neurons at high rates. This advancement has the potential to unlock crucial insights into brain function and lead to breakthroughs in treating neurological disorders. By involving graduate students, engaging in effective outreach and fostering interdisciplinary collaboration, the project aims to shape the future of cognitive science, neuroscience, and education.Two-photon imaging of intracellular calcium in awake behaving animals has been a catalyst for unprecedented advances in the understanding of cognitive neural processes. However, calcium time courses are a noisy convolution of neuronal spiking with calcium efflux dynamics that hide potentially crucial information on subthreshold voltage changes and fast action potential firing. In the last three decades several groups have developed promising new approaches to overcome this inherent disadvantage by imaging neuronal membrane potential using genetically encoded voltage indicators (GEVIs). Imaging GEVIs by a wide neuroscience community would provide information that is lost in calcium imaging potentially yielding crucial breakthroughs in understanding the neural basis of behavior. However, GEVI imaging has not been adopted widely because it suffers from low signal to noise ratio (SNR), limited ability to image large ensembles of neurons and difficult implementation by the scientific community. Here, the investigators will develop a novel large ensemble GEVI imaging technique and will apply it to understand the complex process of cerebellar involvement in rapid decision making. A new approach is proposed for large ensemble GEVI imaging in which two photon microscopy is coupled with adaptive optical microprism technology. The result is beamlet excitation of subsets of neurons, with high imaging rates (500 Hz). The team will continue dissemination efforts with the technology developed in this project. Finally, the investigators will endeavor to communicate science to the broader audience through venues such as the Denver Museum of Nature and Science and CU Science Discovery program.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该研究项目旨在开发一种革命性的显微镜来研究大脑的神经活动。通过结合工程、物理学和神经科学的专业知识，多学科团队正在致力于一项高风险、高回报的努力，以克服现有成像技术的局限性。研究人员正专注于基因编码电压指示器（GEVI）来捕获快速神经元放电和阈下电压变化。这种创新方法将双光子显微镜与电润湿自适应光学微棱镜技术相结合，能够对神经元组进行高速选择性激发和成像。这一进展有可能解锁对大脑功能的重要见解，并在治疗神经系统疾病方面取得突破。该项目旨在通过研究生的参与、有效的推广和促进跨学科合作，塑造认知科学、神经科学和教育的未来。对清醒行为动物细胞内钙的双光子成像已成为推动认知科学、神经科学和教育领域前所未有的进步的催化剂。了解认知神经过程。然而，钙时间进程是神经元尖峰与钙流出动力学的噪声卷积，隐藏了有关亚阈值电压变化和快速动作电位放电的潜在关键信息。在过去的三十年中，一些研究小组开发出了有前景的新方法，通过使用基因编码电压指示器（GEVI）对神经元膜电位进行成像来克服这种固有的缺点。广泛的神经科学界对 GEVI 进行成像将提供钙成像中丢失的信息，从而可能在理解行为的神经基础方面产生关键突破。然而，GEVI 成像尚未得到广泛采用，因为它的信噪比 (SNR) 较低、对大型神经元群成像的能力有限以及科学界难以实施。在这里，研究人员将开发一种新型大型整体 GEVI 成像技术，并将应用它来了解小脑参与快速决策的复杂过程。提出了一种用于大型整体 GEVI 成像的新方法，其中两个光子显微镜与自适应光学微棱镜技术相结合。结果是神经元子集的小束激发，具有高成像速率（500 Hz）。该团队将继续努力传播该项目中开发的技术。 最后，研究人员将努力通过丹佛自然科学博物馆和科罗拉多大学科学发现计划等场所向更广泛的受众传播科学。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值进行评估，被认为值得支持以及更广泛的影响审查标准。