High-Speed, wide field fluorescent imaging of cortex in freely moving animals

自由活动动物皮层的高速、宽视场荧光成像

• 批准号: 8490456
• 负责人: Eugenio Culurciello
• 金额: $ 61.65万
• 依托单位: JOHN B. PIERCE LABORATORY, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8490456
• 关键词: Affect; Algorithms; Amplifiers; Anesthetics; Animal Behavior; Animals; Area; Back; Behavioral; Behavioral Paradigm; Biological; Boxing; Brain; Cells; Cerebral cortex; Collection; Computer Assisted; Computers and Advanced Instrumentation; Conscience; Consumption; Cues; Custom; Data; Data Set; Development; Devices; Discrimination; Dyes; Electrocardiogram; Electroencephalography; Electronics; Elements; Engineering; Ensure; Environment; Event; Fluorescence; Frequencies; Funding; Generations; Goals; Head; Heating; Height; Human; Image; Imaging Device; Laboratories; Light; Lighting; Mammals; Maps; Measures; Membrane; Memory; Methods; Microelectrodes; Microscope; Neurons; Noise; Optics; Output; Paralysed; Patients; Physiologic pulse; Physiology; Process; Property; Prosthesis; Pump; RH1691; Rattus; Records; Records Controls; Resolution; Robotics; Rodent; Running; Sampling; Signal Transduction; Somatosensory Cortex; Source; Speed; Stimulus; Subfamily lentivirinae; Surface; System; Temperature; Testing; Time; Tissues; Training; Translating; Vibrissae; Weight; analog; awake; barrel cortex; brain machine interface; brain tissue; cranium; data mining; design; digital; fluorescence microscope; heat exchanger; in vivo; instrument; lens; light intensity; light weight; mind control; miniaturize; novel; portability; prototype; research study; response; restraint; sample fixation; sensor; voltage; whisker discrimination

DESCRIPTION (provided by applicant): The study of neuronal activity in awake, freely moving animals is the most representative view of 'normal' neuronal function. Anesthetics and restraint have profound effects on neuronal physiology and the correlate animal behavior. The study of real time neuronal event processing requires recording methods with high temporal bandwidth (>500 hz) and the ability to detect physiologically relevant events (i.e. voltage changes). Currently only microelectrode recording of single neuronal units, multiunit activity and field potentials have sufficient temporal resolution and portability to allow recording of neuronal activity in freely moving animals. The proposed project will produce three miniature microscope/imaging systems which can be head-mounted and will allow the optical recording of changes in membrane voltage of neurons in freely moving rats. During the funded two-year ARRA project, two prototype devices and two image sensors were developed that collect wide field image sequences of fluorescent voltage dye signals. The current application will complete the fabrication and operationalize two different style, head-mountable, high-speed, fluorescence microscopes. This includes the design and fabrication of two different finalized, imaging sensors, custom optics for each, a stabilized illumination source and form fit microscope bodies with skull mounts. The studies proposed herein will also build and test a novel fluorescence microscope design that will reduce the height of the traditional microscope by eliminating the 45¿ dichroic mirror. We will also miniaturize our current camera control and recording system to be fully contained in a back pack on the rat. This back pack will contain a field programmable gate array, heat exchanger/coolant pump, SD memory and a battery to run the microscope fully autonomously. Finally, the instruments will be tested in imaging studies of the rat somatosensory cortex (barrel cortex). Cortical responses to object discrimination will be studied in freely moving rats. These studies will systematically engineer each component to maximize excitation light delivery, fluorescent light collection and recording speed while minimizing weight volume, energy usage and heat generation. The device will be capable of recording rapid (1000 fps) fluorescent image sequences of a >2 mm2 area of cortex and detect small changes in ?F/F (0.1%). The device will be small (<1 cm2 head mounted) and light weight enough (5 g) to be mounted on the head of a freely moving rat. The device is intended as a precursor to devices that can translate neuronal activity in humans into actions in real time, optical brain machine prosthetic.

描述（由申请人提供）：对清醒、自由活动的动物的神经元活动的研究是“正常”神经元功能最具代表性的观点。麻醉剂和约束对神经生理学和相关的动物行为具有深远的影响。实时神经元事件处理的研究需要具有高时间带宽（> 500 Hz）的记录方法以及检测生理相关事件（即电压变化）的能力。目前，只有单个神经元单元、多单元活动和场电位的微电极记录具有足够的时间分辨率和可移植性，以允许记录自由移动的动物中的神经元活动。拟议的项目将生产三个微型显微镜/成像系统，这些系统可以安装在头上，并允许光学记录自由移动大鼠神经元膜电压的变化。在资助的为期两年的 ARRA 项目中，开发了两个原型设备和两个图像传感器，用于收集荧光电压染料信号的宽场图像序列。当前的应用程序将完成两种不同风格的头戴式高速荧光显微镜的制造和操作。这包括设计和制造两种不同的最终成像传感器、每种传感器的定制光学器件、稳定的照明源以及带有头骨支架的形状适合的显微镜主体。本文提出的研究还将构建并测试一种新颖的荧光显微镜设计，该设计将通过消除 45° 二向色镜来降低传统显微镜的高度。我们还将小型化我们当前的摄像机控制和记录系统，以完全包含在老鼠的背包中。该背包将包含现场可编程门阵列、热交换器/冷却剂泵、SD 内存和用于完全自主运行显微镜的电池。最后，这些仪器将在大鼠体感皮层（桶状皮层）的成像研究中进行测试。将在自由移动的大鼠中研究皮层对物体辨别的反应。这些研究将系统地设计每个组件，以最大限度地提高激发光传输、荧光收集和记录速度，同时最大限度地减少重量体积、能源消耗和热量产生。该设备将能够记录 >2 mm2 皮质区域的快速 (1000 fps) 荧光图像序列，并检测 ΔF/F (0.1%) 的微小变化。该设备体积小（头戴式<1 cm2）且重量轻（5 g），可以安装在自由活动的老鼠的头上。该设备旨在作为能够将人类神经元活动转化为实时动作的设备的先驱，即光学脑机假肢。