Optimization of 3-photon microscopy for Large Scale Recording in Mouse Brain

用于小鼠大脑大规模记录的三光子显微镜优化

• 批准号: 9130300
• 负责人: CHRIS XU
• 金额: $ 48.32万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9130300
• 关键词: Academia; Area; Biomedical Research; Brain; Brain imaging; Collaborations; Collection; Communities; Dendrites; Deposition; Development; Device or Instrument Development; Elements; Feedback; Fiber; Fluorescence Microscopy; Generations; Genetic Engineering; Goals; Green Fluorescent Proteins; Head; Health; Image; Imaging Device; Individual; Industry; Interdisciplinary Study; Lasers; Lead; Microscope; Microscopy; Mus; Neurobiology; Neurons; Neurosciences; Noise; Optics; Participant; Penetration; Performance; Photons; Physiologic pulse; Procedures; Process; Proteins; Research; Research Personnel; Resolution; Signal Transduction; Source; Speed; System; Technology; Testing; Tissues; Universities; Work; adaptive optics; awake; base; biological research; brain tissue; commercialization; design; experience; fluorophore; improved; in vivo; industry partner; instrument; lens; meetings; performance tests; programs; research and development; tool; two-photon; virtual reality

 DESCRIPTION (provided by applicant): The goal of this research program is to optimize three-photon fluorescence microscopy (3PM) for large scale, noninvasive, volumetric imaging of neuronal activity. To leverage the superb performance of green-fluorescent protein based genetically engineered Ca-probes (e.g., GCaMPs), 3PM at the 1300-nm spectral window will be developed, which not only preserves the tissue penetration capability of 3PM at the longer excitation wavelength but also enables a wide variety of blue and green fluorophores, including a number of fluorescent proteins and Ca-indicators, to be excitable via three-photon excitation. To improve the signal-to-noise ratio (SNR) so that a practical frame rate can be achieved for imaging dynamic brain activity even at a penetration depth of 1.1 mm or beyond, new objective lenses will be designed and fabricated that will collect the signal efficiently at depth. In additin, the lens design will also support convenient integration with adaptive optics (AO), with the goal of making AO a routine imaging tool in a neuroscience lab. To improve both SNR and spatial resolution, AO in 3PM at 1300 nm will be employed. The impact of AO for increasing signal generation is significantly higher for 3PM than 2PM because of the higher order nonlinear process. The impact of AO is also expected to increase with increasing imaging depth. The aim is to achieve close to diffraction limited spatial resolution for 3PM at 1300 nm, which will be sufficient to resolve individual dendrite. A strong interdisciplinary research team has been assembled, including participants from both industry and academia, to perform the research and development. The successful completion of this program will have a broad impact on neuroscience where high-resolution, high speed imaging deep within an intact mouse brain is required.

 描述（由适用提供）：该研究计划的目标是优化三光子荧光显微镜（下午3点），以大规模，无创，无创的，体积成像的神经元活性成像。将开发基于绿色荧光蛋白的遗传学蛋白质的CA探针（例如，GCAMP）的出色性能，将开发1300 nm的光谱窗口下的下午3点，这不仅可以保留较长的令人兴奋的波长的组织渗透能力，还可以通过较长的令人兴奋的波长，还可以通过各种蓝色和绿色的荧光素（包括蓝色和绿色的荧光素）来激发效果，并在包括荧光素的范围内激发效果。三光子兴奋。为了提高信噪比（SNR），以便即使在1.1 mm或更高的穿透深度下，还可以实现动态大脑活动的实用帧速率，将设计和制造新的物镜镜头，以在深度上有效地收集信号。在附加素中，镜头设计还将支持与自适应光学（AO）的方便集成，目的是使AO在神经科学实验室中成为常规成像工具。为了改善SNR和空间分辨率，将在1300 nm处的下午3点的AO雇用。由于较高的非线性过程，AO对增加信号生成的影响明显高于下午2点。预计AO的影响随着成像深度的增加而增加。目的是在1300 nm处达到3pm的衍射有限的空间分辨率，这足以解决单个树突。一支强大的跨学科研究团队已经组建了，包括来自行业和学术界的参与者，以执行研究与开发。该程序的成功完成将对神经科学产生广泛的影响，因为需要高分辨率，高速成像在完整的小鼠大脑内。