A multi-foci objective lens for large scale brain activity recording

用于大规模大脑活动记录的多焦点物镜

• 批准号: 10731905
• 负责人: CHRIS XU
• 金额: $ 32.8万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10731905
• 关键词: 3-Dimensional; Area; Back; Behavior; Brain; Brain imaging; Brain region; Cancer Biology; Collection; Complex; Confocal Microscopy; Fluorescence; Goals; Hippocampus; Image; Immunology; Light; Microscope; Microscopy; Modality; Mus; Neurons; Neurosciences; Optical Methods; Optics; Pathway interactions; Pattern; Penetration; Photons; Physiologic pulse; Population; Resolution; Sampling; Scanning; Signal Transduction; Speed; Technology; Thinness; Time; Tissue imaging; Tissues; brain tissue; brain volume; design; fabrication; high resolution imaging; imaging modality; improved; in vivo; in vivo imaging; lens; meter; multi-photon; neural circuit; neuronal circuitry; novel; optical imaging; programs; three photon microscopy; timeline; two photon microscopy; two-photon

Abstract The goal of this project is to develop a novel bifocal catadioptric objective that will allow large-scale recording of neural circuits in vivo. The objective will enable faster volumetric imaging of large brain regions by simultaneous two-photon (2P) imaging of shallow layers and three-photon (3P) imaging of deep layers of brain tissues with improved collection efficiency. Although three-photon microscopy (3PM) allows imaging at depths inaccessible by two photon microscopy (2PM), 2P excitation generates larger fluorescence signal with lower excitation pulse energy when imaging at shallow tissue layers. Therefore, for fast imaging across a large depth, the optimum approach is to use 2PM for the superficial layers and 3PM for the deeper regions simultaneously. Implementation of this approach inevitably requires the objective lens to generate two focal planes that are separated by a large axial distance while still maintaining high spatial resolution and large field of view. Simultaneous 2PM and 3PM will not only allow for utilization of the advantages of both modalities but also for faster volumetric imaging of large brain columns. We will develop a bifocal catadioptric (i.e., both refractive and reflective) lens based on the idea of separation the optical paths of the excitation light with different wavelengths. The lens will feature two focal planes separated axially by ~ 600 µm for the 2P (< 1100 nm) and 3P (>1200 nm) excitation wavelengths. The design approach also separates the excitation path and the collection path and allows independent optimization for efficient collection of the emitted fluorescence. The bifocal objective will collect fluorescence back through non-imaging pathways, which enables the proposed catadioptric objective to have a large collection numerical aperture and a large collection field of view. The collection efficiency is approximately 5x higher than the commercially available objective lenses when imaging deep (>1 mm) into the mouse brain. Improving the signal collection efficiency will immediately increase the frame rate without increasing the excitation power, enabling high-resolution, high-speed imaging at these depths. We will design, fabricate and validate the novel objective lens and will combine it with focus-tunable lenses to enable faster volumetric imaging of mouse brains. The successful completion of this program will immediately enable simultaneous 2P and 3P imaging across a large range of depth (~ 1.2 mm), such as recording population of neurons across different layers of mouse brains. The technology developed within this program will have potential impacts in a large number of biomedical fields such as neuroscience, immunology, and cancer biology.

摘要 这个项目的目标是开发一种新的双焦点折反射物镜，将允许大规模记录的 体内的神经回路该目标将能够通过同时对大脑大区域进行更快的体积成像， 浅层的双光子（2 P）成像和脑组织深层的三光子（3 P）成像， 提高收集效率。虽然三光子显微镜（3 PM）可以在无法到达的深度成像， 在双光子显微镜下，2 P激发在较低的激发脉冲下产生较大的荧光信号 在浅组织层成像时的能量。因此，对于跨越大深度的快速成像， 方法是同时使用2PM用于浅层和3 PM用于较深区域。执行 这种方法的一个优点是不可避免地要求物镜透镜产生两个焦平面， 轴向距离，同时仍然保持高空间分辨率和大视场。同时2PM和3 PM 将不仅允许利用两种模式的优点，而且还允许更快的体积成像， 巨大的脑柱我们将开发一种双焦点折反射（即，折射和反射）的透镜 分离具有不同波长的激发光的光路的想法。该透镜将采用两个 对于2 P（< 1100 nm）和3 P（>1200 nm）激发波长，焦平面轴向间隔约600 µm。 该设计方法还分离了激发路径和收集路径，并允许独立的 优化以有效收集发射的荧光。双焦点物镜将收集荧光 通过非成像路径返回，这使得所提出的折反射物镜能够具有大的收集 数值孔径和大的收集视场。收集效率约为5倍， 当成像深入（>1 mm）小鼠大脑时，市售物镜。提高 信号收集效率将立即增加帧速率而不增加激励功率， 能够在这些深度进行高分辨率、高速成像。我们将设计，制造和验证小说 物镜透镜，并将联合收割机与可调焦透镜相结合，以实现小鼠大脑的更快体积成像。 该计划的成功完成将立即实现跨多个区域的同时2 P和3 P成像。 大范围的深度（~ 1.2 mm），例如记录小鼠大脑不同层的神经元群体。 该计划开发的技术将在大量生物医学领域产生潜在影响 如神经科学、免疫学和癌症生物学。