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革命性的显微镜研究大脑的神经活动。通过结合工程，物理和神经科学的专业知识，多学科团队正在研究高风险，高付费的努力，以克服现有成像技术的局限性。研究人员着重于遗传编码的电压指标（GEVIS），以捕获快速的神经元放电和亚阈值电压变化。创新的方法将两光子显微镜与电视自适应光学微普罗普技术结合起来，从而可以以高速率对神经元组的选择性激发和成像进行成像。这种进步有可能解开对脑功能的关键见解，并在治疗神经系统疾病方面取得突破。通过参与研究生，进行有效的外展活动并促进跨学科的合作，该项目旨在塑造认知科学，神经科学和教育的未来。对清醒行为的动物中细胞内钙的两次化合物成像一直是对认知神经过程的前所未有的预言的催化剂。但是，钙时课程是神经元尖峰的嘈杂卷积，钙外排动力学隐藏了有关亚阈值电压变化和快速动作电位射击的潜在至关重要的信息。在过去的三十年中，几个小组通过使用遗传编码的电压指标（GEVIS）对神经元膜电位（GEVIS）进行成像，开发了有希望的新方法来克服这种固有的劣势。广泛的神经科学界的成像Gevis将提供在钙成像中丢失的信息，这可能会在理解行为的神经基础上产生至关重要的突破。但是，GEVI成像之所以被广泛采用，是因为它遭受了低信号与噪声比（SNR）的损害，图像有限的图像神经元的大集合以及科学界的艰难实施能力。在这里，研究人员将开发一种新型的大型集合GEVI成像技术，并将其应用于小脑参与快速决策的复杂过程。提出了一种针对大型集成GEVI成像的新方法，其中两个光子显微镜与自适应光学微普罗普技术耦合。结果是神经元子集的Beamlet激发，具有高成像率（500 Hz）。该团队将继续在该项目中开发的技术继续传播工作。 最后，调查人员将通过诸如丹佛自然与科学博物馆和CU科学发现计划等场所努力将科学传达给更广泛的受众。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛影响标准通过评估来通过评估来支持的。