Toward fast and deep imaging of living tissue with cellular resolution

以细胞分辨率对活体组织进行快速、深度成像

基本信息

  • 批准号:
    10651713
  • 负责人:
  • 金额:
    $ 62.33万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2022
  • 资助国家:
    美国
  • 起止时间:
    2022-07-01 至 2026-03-31
  • 项目状态:
    未结题

项目摘要

Abstract An exciting recent development for high spatial resolution deep tissue imaging is long wavelength three- photon fluorescence microscopy (3PM). Since its first demonstration of imaging subcortical structures in the mouse brain, 3PM has driven rapid progress in deep tissue imaging beyond the depth limit of two-photon fluorescence microscopy (2PM). Long-wavelength 3PM is perhaps the most promising new technology for deep imaging within scattering biological tissues, and has potential impacts in a large number of biomedical fields such as neuroscience, immunology, and cancer biology. On the other hand, there are a number of challenges that must be overcome before 3PM can reach its full potential. Because it is a higher-order nonlinear process, three- photon excitation (3PE) is inherently weaker than two-photon excitation (2PE). The weak signal strength of 3PM is particularly problematic for fast imaging of dynamic cellular process. Furthermore, the laser sources for 3PM are not yet optimized for deep tissue penetration, and the complexity and cost of the excitation source is a major barrier for the applications of 3PM in a typical biomedical research lab. Finally, nearly all 3PM applications today are in the brains. Reaching anatomical frontiers is equally possible in other organs with 3PM, but explicit demonstrations of intravital imaging in novel locations are needed to bring deep imaging capability to other biological systems. The research activity of this proposal will directly address the above challenges for in vivo deep tissue 3PM. We will develop a new generation of 3PM that will improve the performance of existing 3PM by two orders of magnitude and enable multi-color deep tissue imaging with a single excitation wavelength. We will demonstrate the unprecedented imaging capabilities with a low-cost, fiber-based laser system, removing a key barrier for the deployment of 3PM in biology labs. Furthermore, by applying our techniques to a wide variety of biological systems, we will create a valuable knowledge base for the applications of 3PM. Our development of the next generation 3PM parallels the development of 2PM, where the concerted development effort in lasers, microscopes, and biological applications in the 1990s made 2PM ubiquitous in biomedical research labs by the early 2000s. Our vision is to make deep, fast 3PM a routine instrument for a wide variety of biomedical applications just as 2PM does in the shallower regions of biological tissues and organs. The successful completion of this program will enable visualization of dynamic process at the sub-cellular level in intact organs and animal models that are completely beyond the reach of any existing imaging techniques.
摘要 用于高空间分辨率深部组织成像的令人兴奋的最新发展是长波长三波长成像。 光子荧光显微镜(3 PM)。自从它首次展示了大脑皮层下结构的成像以来, 小鼠大脑,3 PM推动了超越双光子深度限制的深层组织成像的快速进展 荧光显微镜(2PM)。长波3 PM也许是最有前途的新技术, 在散射生物组织内成像,并且在大量生物医学领域具有潜在的影响, 神经科学、免疫学和癌症生物学。另一方面,也有一些挑战, 在3 PM充分发挥其潜力之前,必须克服这些障碍。因为它是一个高阶非线性过程,三个- 光子激发(3 PE)固有地弱于双光子激发(2 PE)。下午3点的微弱信号 对于动态细胞过程的快速成像尤其成问题。此外,用于3 PM的激光源 尚未针对深层组织穿透进行优化,并且激发源的复杂性和成本是主要的 在典型的生物医学研究实验室中应用3 PM的障碍。最后,今天几乎所有的3 PM应用程序 都在大脑里在其他器官中,3 PM同样可能到达解剖学前沿,但明确 需要在新的位置进行活体成像的演示,以将深度成像能力带给其他 生物系统。本提案的研究活动将直接解决上述体内挑战 深层组织下午3点。我们将开发新一代3 PM,以提高现有3 PM的性能 通过两个数量级,并且能够利用单个激发波长进行多色深层组织成像。我们 将通过低成本、基于光纤的激光系统展示前所未有的成像能力,消除了 在生物实验室中部署3 PM的关键障碍。此外,通过将我们的技术应用于各种各样的 我们将为3 PM的应用创造一个有价值的知识基础。我们的发展 下一代3 PM的发展与2PM的发展并行,在2PM中,激光器的协调发展努力, 20世纪90年代,显微镜和生物学应用使得2PM在生物医学研究实验室中无处不在。 2000年代初我们的愿景是使深度、快速的3 PM成为各种生物医学的常规仪器 就像2PM在生物组织和器官的较浅区域中的应用一样。成功 该项目的完成将使完整器官在亚细胞水平上的动态过程可视化 和动物模型,这完全超出了任何现有的成像技术的范围。

项目成果

期刊论文数量(1)
专著数量(0)
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专利数量(0)

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Nozomi Nishimura其他文献

Nozomi Nishimura的其他文献

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{{ truncateString('Nozomi Nishimura', 18)}}的其他基金

Novel tracers for in vivo studies of waste transport by fluid flows in the brain
用于脑内液体流动废物运输体内研究的新型示踪剂
  • 批准号:
    10732612
  • 财政年份:
    2023
  • 资助金额:
    $ 62.33万
  • 项目类别:
Simultaneous, Cell-Resolved, Bioluminescent Recording From Microcircuits
微电路同步、细胞解析、生物发光记录
  • 批准号:
    10463819
  • 财政年份:
    2021
  • 资助金额:
    $ 62.33万
  • 项目类别:
Simultaneous, Cell-Resolved, Bioluminescent Recording From Microcircuits
微电路同步、细胞解析、生物发光记录
  • 批准号:
    10294095
  • 财政年份:
    2021
  • 资助金额:
    $ 62.33万
  • 项目类别:
Stalled capillary flow: a novel mechanism for hypoperfusion in Alzheimer disease
毛细血管血流停滞:阿尔茨海默病低灌注的新机制
  • 批准号:
    10412670
  • 财政年份:
    2021
  • 资助金额:
    $ 62.33万
  • 项目类别:
Age Compromises Novel Motility and Repair Functions in Stem Cell Niche of Intestinal Crypts
年龄会损害肠隐窝干细胞生态位的新活力和修复功能
  • 批准号:
    9753843
  • 财政年份:
    2018
  • 资助金额:
    $ 62.33万
  • 项目类别:
Diffuse, spectrally-resolved optical strategies for detecting activity of individual neurons from in vivo mammalian brain with GEVIs
使用 GEVI 检测体内哺乳动物大脑中单个神经元活动的漫反射光谱分辨光学策略
  • 批准号:
    9395599
  • 财政年份:
    2017
  • 资助金额:
    $ 62.33万
  • 项目类别:
In vivo tools for analyzing interstitial fluid flow
用于分析间质液流动的体内工具
  • 批准号:
    9751865
  • 财政年份:
    2017
  • 资助金额:
    $ 62.33万
  • 项目类别:
Supplement: Stalled capillary flow affects protein clearance by modulating interstitial fluid flow
补充:毛细血管血流停滞通过调节间质液流动影响蛋白质清除
  • 批准号:
    10617575
  • 财政年份:
    2015
  • 资助金额:
    $ 62.33万
  • 项目类别:
Role of Microvascular Lesions in Alzheimer's Disease
微血管病变在阿尔茨海默病中的作用
  • 批准号:
    8140740
  • 财政年份:
    2010
  • 资助金额:
    $ 62.33万
  • 项目类别:
Role of Microvascular Lesions in Alzheimer's Disease
微血管病变在阿尔茨海默病中的作用
  • 批准号:
    8044027
  • 财政年份:
    2010
  • 资助金额:
    $ 62.33万
  • 项目类别:

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