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 这项工作的目的是使用在可植入的，纤维耦合显微镜中的微型扫描镜进行移动，行为小鼠的3D脑体积的高速光学荧光成像。现有用于神经成像的可植入显微镜主要对单个焦平面进行成像，或者使用缓慢和/或减少光学分辨率的扫描机制改变图像深度。基于微电机电系统（MEMS）技术的扫描镜可以放置在光纤耦合的微型显微镜的客观镜头附近，并达到非常高的扫描频率。基于主动压电材料的MEMS扫描仪可以在空间中的所有三个轴上迅速转向激光光。然后，使用超快激光激发的多光子效应可以通过深层成像穿透进行荧光成像。我们的具体目的是（1）展示具有适合脑成像的速度和外侧扫描的高性能MEM镜子； （2）将镜子整合到原型可植入显微镜中，以在细胞到细胞分辨率下进行3D成像； （3）从移动小鼠那里收集神经激活的图像。小镜子质量和主动扫描仪控制将防止不希望的运动伪像，并增加图像体积中感兴趣的本地化区域的采样频率。这项工作的成功结论将在长时间的3D大脑环境中为神经元行为提供新的工具，这是研究神经功能（包括运动，社会行为和学习）所必需的。