Ultrafast high-contrast voltage imaging in freely moving animals

自由移动动物的超快高对比度电压成像

• 批准号: 10445419
• 负责人: Jerome Mertz
• 金额: $ 65.51万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-15 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10445419
• 关键词: Adoption; Anatomy; Animals; Area; Behavior; Brain; Cells; Communities; Confocal Microscopy; Coupled; Data; Dendrites; Detection; Development; Fiber; Generations; Goals; Head; Image; Individual; Label; Lateral; Light; Lighting; Methods; Microscope; Microscopy; Mus; Neurons; Neurosciences; Noise; Outcome; Performance; Population; Positioning Attribute; Resolution; Scanning; Side; Signal Transduction; Social Interaction; Speed; System; Time; Tissues; animal imaging; awake; base; behavioral study; contrast imaging; flexibility; imaging platform; imaging probe; imaging system; improved; instrument; interest; millisecond; optical imaging; prototype; relating to nervous system; spatiotemporal; theories; tool; voltage; way finding

ABSTRACT We propose to develop a targeted illumination confocal (TICO) microscope to enable high speed, large-scale voltage imaging in the brain. This microscope will be based on the combination of two key strategies. The first strategy is high-speed confocal microscopy based on line scanning. The benefit of confocal microscopy is that out-of-focus background is largely rejected by the use of slit detection. We will supplement this background rejection with the additional strategy of targeted illumination. In this way, we will confine our excitation light only to pre-selected regions of interests (e.g., cell bodies or dendrites), minimizing the generation of out-of-focus background in the first place. These strategies will be implemented with the use of a double-sided scanning mechanism that will enable us to perform fast scanning to fully exploit the ultrafast throughput and remarkably low noise capabilities of state-of-the-art CMOS cameras. By combining the complementary advantages of confocal detection and targeted illumination, we expect to significantly improve the contrast and SNR of our voltage imaging relative to the current methods based on cameras. Our specific aims are to build two dual-region TICO microscopes, one adapted for head-fixed animal imaging, and the other adapted for freely moving animal imaging using a miniature microscope objective coupled by a flexible fiber bundle. Establishing the feasibility of TICO microscopy in freely moving animals would constitute a major breakthrough and will lay the groundwork for future research into the relations between voltage activity and behavior in natural conditions. The ability to image freely moving animals will be indispensable for this goal.

抽象的 我们建议开发有针对性的照明共焦（TICO）显微镜，以使高速， 大脑中的大型电压成像。该显微镜将基于两个的组合 关键策略。第一个策略是基于线条扫描的高速共聚焦显微镜。这 共聚焦显微镜的好处是，使用狭缝在很大程度上被拒绝 检测。我们将通过针对性的其他策略来补充这种背景拒绝 照明。这样，我们将仅将激发光限制在预先选择的兴趣区域 （例如，细胞体或树突），最大程度地减少了第一个焦点背景的产生 地方。这些策略将通过使用双面扫描机制来实施 这将使我们能够进行快速扫描以完全利用超快吞吐量并明显地 最先进的CMOS摄像机的低噪声能力。通过结合补充 共聚焦检测和有针对性照明的优点，我们希望显着改善 相对于基于相机的当前方法，我们的电压成像的对比度和SNR。 我们的具体目的是构建两个双区域TICO显微镜，一个适用于头部固定 动物成像，另一个适用于使用微型的自由移动动物成像 显微镜物镜由柔性纤维束结合。建立TICO的可行性 自由移动动物的显微镜将构成一个重大突破，并将 将来研究电压活动与自然行为之间关系的基础 状况。对于这个目标，可以自由移动动物的能力是必不可少的。

项目成果

Large-scale deep tissue voltage imaging with targeted illumination confocal microscopy.

使用靶向照明共聚焦显微镜进行大规模深层组织电压成像。

• DOI: 10.1101/2023.07.21.548930
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Xiao,Sheng;Cunningham,WilliamJ;Kondabolu,Krishnakanth;Lowet,Eric;Moya,MariaV;Mount,Rebecca;Ravasio,Cara;Economo,MichaelN;Han,Xue;Mertz,Jerome
• 通讯作者: Mertz,Jerome