Implantable 3D fluorescence imaging with high-speed, addressable laser scanning in moving mice

通过高速、可寻址激光扫描对移动小鼠进行植入式 3D 荧光成像

• 批准号: 10614795
• 负责人: Kenn R Oldham
• 金额: $ 169.69万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614795
• 关键词: 3-Dimensional; Acceleration; Action Potentials; Animals; Area; Articular Range of Motion; Behavior; Brain; Brain imaging; Calcium; Collection; Coupled; Coupling; Dimensions; Distal; Environment; Feedback; Fiber; Fiber Optics; Film; Frequencies; Future; Geometry; Goals; Height; Image; Imaging Device; Implant; Individual; Lasers; Lateral; Learning; Lens Fiber; Light; Lighting; Location; Locomotion; Longitudinal Studies; Mammals; Memory; Microscope; Monitor; Morphologic artifacts; Motion; Movement; Mus; Neurons; Neurophysiology - biologic function; Operative Surgical Procedures; Optics; Penetration; Performance; Positioning Attribute; Resolution; Sampling; Scanning; Signal Transduction; Social Behavior; Social Interaction; Specific qualifier value; Speed; System; Thinness; Three-Dimensional Imaging; Time; Training; Validation; Work; brain volume; calcium indicator; design; energy density; experimental study; fluorescence imaging; high resolution imaging; implantation; instrument; interest; microsystems; millisecond; multi-photon; neural; neuroimaging; novel; optical fiber; optical imaging; optogenetics; prevent; prototype; response; technology platform

Project Summary The goal of this work is to achieve high-speed optical fluorescence imaging of 3D brain volume in moving, behaving mice, using miniature scanning mirrors in an implantable, fiber-coupled microscope. Existing implantable microscopes for neural imaging primarily image a single focal plane or may alter image depth using scanning mechanisms that are slow and/or reduce the optical resolution. Scanning mirrors based on microelectromechanical system (MEMS) technology can be placed near the objective lens of fiber-coupled miniature microscopes, and achieve very high scanning frequencies. MEMS scanners based on active piezoelectric materials can rapidly steer laser light through all three axes in space. Fluorescence imaging can then be performed with deep imaging penetration using the multi-photon effect excited by an ultrafast laser. Our Specific Aims are to (1) demonstrate high-performance MEMS mirrors for axial and lateral scanning with speed and form factor appropriate for brain imaging; (2) integrate mirrors into a prototype implantable microscope, for 3D imaging at cellular-to-sub-cellular resolution; (3) collect images of neural activation from moving mice. Small mirror mass and active scanner control will prevent undesired motion artifacts and increase sampling frequencies in localized regions of interest within the image volume. The successful conclusion of this work would provide novel instruments for neuron behavior in the 3D brain environment over extended periods, necessary for the study of neural functions including locomotion, social behavior, and learning.

项目摘要 这项工作的目标是实现高速光学荧光成像的三维脑体积在移动，行为小鼠，使用微型扫描镜在可植入的，光纤耦合显微镜。现有的用于神经成像的可植入显微镜主要对单个焦平面成像，或者可以使用缓慢和/或降低光学分辨率的扫描机制来改变图像深度。基于微机电系统（MEMS）技术的扫描镜可以放置在光纤耦合微型显微镜的物镜透镜附近，并实现非常高的扫描频率。基于有源压电材料的MEMS扫描仪可以快速引导激光通过空间中的所有三个轴。然后可以使用由超快激光激发的多光子效应进行具有深度成像穿透的荧光成像。我们的具体目标是：（1）展示高性能MEMS反射镜，用于轴向和横向扫描，速度和形状适合大脑成像;（2）将反射镜集成到原型植入式显微镜中，用于细胞到亚细胞分辨率的3D成像;（3）收集移动小鼠的神经激活图像。小的反射镜质量和主动扫描仪控制将防止不期望的运动伪影，并增加图像体积内的局部感兴趣区域中的采样频率。这项工作的成功完成将为3D大脑环境中的神经元行为提供新的工具，这对于研究神经功能（包括运动，社会行为和学习）是必要的。