Real-time Aberration Sensor for Large-Scale Microscopy Deep in the Mouse and Adult Zebrafish Brain

用于小鼠和成年斑马鱼大脑深处的大规模显微镜检查的实时像差传感器

基本信息

批准号：
10166305
负责人：
Steven Graham Adie
金额：
$ 198.23万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10166305
关键词：
Address Adult Algorithms Biological Biological Process Brain Brain Diseases Brain imaging Brain region Budgets Calibration Cells Cerebellum Consumption Financial compensation Fluorescence Goals Hybrids Image Imaging Techniques Imaging technology Individual Label Lasers Maps Methods Microscope Microscopy Modality Morphologic artifacts Motion Mus Neurons Neurosciences Neurosciences Research Noise Optical Coherence Tomography Optics Penetration Phase Photobleaching Photons Phototoxicity Physiologic pulse Population Positioning Attribute Procedures Pupil Research Resolution Sampling Shapes Signal Transduction Source Specificity Speed Structure System Techniques Technology Time Tissues Training Zebrafish adaptive optics awake base biological systems brain tissue data acquisition dentate gyrus digital imaging approach imaging study imaging system in vivo interest microscopic imaging mouse model multimodality multiphoton microscopy novel optical imaging parallel computer parallel processing programs relating to nervous system sensor spatiotemporal superior colliculus Corpora quadrigemina three photon microscopy two photon microscopy

项目摘要

ABSTRACT Optical imaging holds tremendous promise in our endeavor to understand brain functions. The major challenges for optical brain imaging are depth and speed. Due to optical aberrations and tissue scattering, the penetration depth and imaging speed of optical microscopy in the brains (e.g., mouse) is limited. The constraints in depth and speed make large-scale, deep imaging of mouse brain activity out of reach of current imaging techniques. Hardware adaptive optics (AO) has proven to be valuable for in vivo brain imaging with two-photon microscopy (2PM), and will have even larger impact for deep brain 3-photon microscopy (3PM); however, existing AO techniques require iterative optimization using multiphoton excited fluorescence signal. While adequate for imaging relatively shallow regions of the brain (< 1 mm deep), iterative optimization is impractical with ultra-deep imaging since the fluorescence signal deceases exponentially with imaging depth. The required integration time to obtain the necessary signal-to-noise ratio for iterative optimization becomes prohibitively long. In this program, we will leverage the advantages provided by computational adaptive optics (CAO) in optical coherence microscopy (OCM), specifically the strong OCT signal (from linear backscattering), and parallel computing on a high-end graphics processing unit (GPU), to provide orders of magnitude speed-up for the sensing of sample-induced aberrations throughout a volume of interest. A long-wavelength OCM system and CAO aberration sensor will be utilized to drive a hardware AO system to correct depth-dependent aberrations, push 3PM imaging depth beyond currently demonstrated limits, and increase imaging speed by at least one order of magnitude. Additionally, by combining with recently developed adaptive excitation laser technology, we will achieve approximately 300-fold increase in photon budget, which will enable truly unprecedented 3PM imaging speed and depth. Successful completion of this program will enable rapid aberration sensing at the depth range of 1 to 2 mm, and will open the exciting new opportunity of recording the neural activity of the dentate gyrus of adult mice through the intact brain. With its high-speed and deep-tissue aberration sensing capability, and zero additional photobleaching and phototoxicity, our novel method for real-time aberration sensing and correction is ideally positioned to transform our ability for large-scale, deep recording of mouse and adult zebrafish brain activity. This imaging approach will significantly extend the information available for neuroscience studies on individual cell-cell as well as circuit interactions that underlie normal and diseased brain function. The technology developed by this proposal will be applicable to imaging in other biological systems where large-scale, deep imaging at high spatiotemporal resolution is needed.

抽象的光学成像在我们的努力中具有巨大的希望，可以理解大脑功能。主要挑战对于光学大脑成像是深度和速度。由于光学畸变和组织散射，穿透大脑中光学显微镜的深度和成像速度（例如，小鼠）受到限制。限制深入速度使小鼠脑活动的大规模成像超出了当前成像技术的范围。硬件自适应光学元件（AO）已被证明对使用两光子显微镜的体内脑成像很有价值（下午2点），对深脑3光子显微镜（3pm）的影响甚至更大；但是，现有的AO 技术需要使用多光子激发荧光信号进行迭代优化。虽然足够了成像相对较浅的大脑区域（深<1毫米），迭代优化是不切实际的，超深成像，因为荧光信号随成像深度呈指数式脱轴。所需的集成时间为了获得迭代优化的必要信号噪声比，长期很长。在这个程序中，我们将利用计算自适应光学器件（CAO）在光学连贯性方面提供的优点显微镜（OCM），特别是强烈的OCT信号（来自线性反向散射），并在A上平行计算高端图形处理单元（GPU），为传感提供速度样本引起的畸变整个兴趣。长波长OCM系统和CAO 畸变传感器将用于驱动硬件AO系统以纠正深度依赖性畸变，推动 3pm成像深度超出当前显示的限制，并将成像速度提高至少一个阶震级。此外，通过与最近开发的自适应激发激光技术相结合，我们将实现大约300倍的光子预算，这将使真正前所未有的3pm成像实现速度和深度。该程序的成功完成将使深度范围内快速畸变感测 1至2毫米，并将为记录齿状回的神经活动的激动人心的新机会成年小鼠通过完整的大脑。具有高速和深层的畸变感应能力，零其他光漂白和光毒性，我们实时畸变感应和校正的新方法是理想的位置可以改变我们对小鼠和成人斑马鱼大脑大规模，深度记录的能力活动。这种成像方法将大大扩展有关神经科学研究的信息单个细胞电池以及脑功能正常和患病的电路相互作用。技术该提案开发的需要在高时空分辨率下进行成像。