A replacement multiphoton microscope for in vivo imaging in rodent models of neur

用于神经啮齿动物模型体内成像的替代多光子显微镜

• 批准号: 7595490
• 负责人: Brian J Bacskai
• 金额: $ 49.15万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595490
• 关键词: Alzheimer' s Disease; Arts; Brain; Brain imaging; Cell Culture Techniques; Dendrites; Detection; Ensure; Environment; Epilepsy; Fluorescence; Fluorescence Resonance Energy Transfer; Functional disorder; Funding; Image; In Vitro; Individual; Life; Measurement; Microscope; Microscopy; Modeling; Neurodegenerative Disorders; Neuroglia; Neurons; Optics; Pathology; Physiologic pulse; Population; Proteins; Quantitative Microscopy; Research; Research Personnel; Resolution; Resources; Rodent Model; Stroke; Structure; System; Techniques; Tissues; United States National Institutes of Health; Vertebral column; Width; experience; in vivo; innovation; instrument; mouse model; nervous system disorder; programs; public health relevance; success

DESCRIPTION (provided by applicant): Multiphoton microscopy is a powerful technique for high resolution intravital imaging in deep tissue, with particularly exciting results obtained from brain imaging. We have successfully implemented and innovated in vivo brain imaging over the past 10 years, most notably in the study of Alzheimer's disease mouse models. These innovations, however, have extended to other brain imaging applications around the world where we have either directly or indirectly assisted in advancing the applications. Our application of multiphoton microscopy began with the first commercial multiphoton microscope, the Biorad 1024MP, which is still functional in the lab, but antiquated. There is no upgrade path for this instrument: it needs to be replaced. We are proposing to purchase a state-of-the-art multiphoton microscopy system to replace this instrument. The configuration of the microscope platform chosen is optimized for fast and deep in-vivo imaging. Using negative chirp optics to compensate for group velocity dispersion in the optical path, the proposed instrument allows optimized pulse-widths for deep tissue imaging with reduced average power. We have assembled a group of experienced, NIH-funded investigators with applications in a variety of neurological diseases that will take advantage of the capabilities of the instrument in an environment rich in experience and proven success. We have been using multiphoton microscopy to image structure and function in vivo on spatial scales as small as a single spine on an identified dendrite from a single neuron. We have developed approaches to image the pathology associated with Alzheimer's disease, neurons, glia, and the vasculature. We have also been using fluorescence lifetime imaging microscopy (FLIM) for sensitive detection of fluorescence resonance energy transfer (FRET), which can be used to reveal interactions between individual proteins. While FLIM has been largely relegated to in vitro or cell culture measurements, we aim to apply FLIM for quantitative imaging in populations of neurons and glia in vivo. The new microscope will be a vital resource for NIH funded imaging studies in mouse models of neurodegenerative diseases, stroke, and epilepsy. PUBLIC HEALTH RELEVANCE: We propose to replace an old multiphoton microscope with a newer more advanced model. We have been using multiphoton microscopy for 10 years to try to understand the pathophysiology in the brain of living mouse models of neurodegenerative disease, and a new microscope will ensure that we can continue our successful research programs.

描述（由申请人提供）：多光子显微镜是一种用于深部组织高分辨率活体成像的强大技术，从脑成像中获得了特别令人兴奋的结果。在过去的10年里，我们成功地实施和创新了体内脑成像，最引人注目的是在阿尔茨海默病小鼠模型的研究中。然而，这些创新已经扩展到世界各地的其他脑成像应用，我们直接或间接地帮助推进这些应用。我们对多光子显微镜的应用始于第一台商用多光子显微镜Biorad 1024MP，该显微镜在实验室中仍能正常工作，但已经过时。此仪器没有升级路径：需要更换。我们建议购买一个最先进的多光子显微镜系统，以取代这一仪器。所选显微镜平台的配置针对快速和深度体内成像进行了优化。使用负啁啾光学补偿光路中的群速度色散，所提出的仪器允许优化的脉冲宽度，以降低平均功率的深部组织成像。我们已经召集了一批经验丰富的NIH资助的研究人员，他们在各种神经系统疾病中应用，这些疾病将在经验丰富和成功证明的环境中利用仪器的功能。我们一直在使用多光子显微镜图像的结构和功能在体内的空间尺度小到一个单一的脊椎上确定的树突从一个单一的神经元。我们已经开发出方法来成像与阿尔茨海默病，神经元，神经胶质细胞和脉管系统相关的病理。我们也一直在使用荧光寿命成像显微镜（FLIM）的荧光共振能量转移（FRET），它可以用来揭示个别蛋白质之间的相互作用的灵敏检测。虽然FLIM已在很大程度上降级到体外或细胞培养测量，我们的目标是应用FLIM在体内的神经元和神经胶质细胞的群体的定量成像。新的显微镜将是NIH资助的神经退行性疾病、中风和癫痫小鼠模型成像研究的重要资源。公共卫生相关性：我们建议用更新更先进的型号取代旧的多光子显微镜。10年来，我们一直在使用多光子显微镜，试图了解神经退行性疾病活体小鼠模型大脑中的病理生理学，新的显微镜将确保我们能够继续我们成功的研究计划。

项目成果

Therapeutic Strategies to Target Calcium Dysregulation in Alzheimer's Disease.

• DOI: 10.3390/cells9112513
• 发表时间: 2020-11-20
• 期刊: Cells
• 影响因子: 6
• 作者: Calvo-Rodriguez M;Kharitonova EK;Bacskai BJ
• 通讯作者: Bacskai BJ

An acute functional screen identifies an effective antibody targeting amyloid-β oligomers based on calcium imaging.

• DOI: 10.1038/s41598-018-22979-2
• 发表时间: 2018-03-15
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Wang X;Kastanenka KV;Arbel-Ornath M;Commins C;Kuzuya A;Lariviere AJ;Krafft GA;Hefti F;Jerecic J;Bacskai BJ
• 通讯作者: Bacskai BJ