Three-Photon Microscope for High-Resolution Deep Tissue Imaging

项目摘要 三光子显微镜（3 PM）是一种光学成像技术，能够在一个特定的环境中进行更深的成像。 散射生物组织比目前领先的标准双光子显微镜（2PM），同时提供 附加的对比度机制和增强的空间分辨率。这项技术的进步使之成为可能 长波长、可调谐、高脉冲能量飞秒激光器和2PM中的相应修改 设置，以适应一种新的激励制度。 作为这些进步的直接结果，啮齿动物大脑中的3 PM成像深度相对于 2PM成像深度-从~700 μm到超过1，200 μm。由于如此大的改进，我们预计 3 PM将允许结构的突破性研究（例如，血管、神经胶质和神经结构）， 功能（例如，氧合、钙信号传导和细胞相互作用），在临床前体内研究中， 在以前难以接近的组织层，如皮质下白色物质和 海马体。本提案中列出的11个NIH资助的项目将从这些成像中受益匪浅 能力的这些项目涵盖了与NIMH使命相关的各种研究主题，包括 提高临床成像模式的定量性，更好地了解 参与各种大脑状况发展的机制。 Martinos生物医学成像中心和马萨诸塞州总医院组成了一个多样化的 研究人员的社区，其中许多人是由美国国立卫生研究院的支持资助，进行尖端的生物医学 这些研究将进一步推进我们对人类健康和疾病的基本理解。这一拟议 3 PM成像系统将代表波士顿地区（可能是东北部）的第一台同类仪器 US），该软件在成像核心中可用。这种尖端仪器将使我们的研究团体 大大加强与NIMH使命相关的各种研究，这也将使马蒂诺 中心继续致力于成为先进生物医学成像的领导者。

Project Summary Three-photon microscopy (3PM) is an optical imaging technology that enables significantly deeper imaging in a scattering biological tissue than the current leading standard two-photon microscopy (2PM), while providing additional contrast mechanisms and enhancing spatial resolution. This technology is made possible by advances in long-wavelength, tunable, high-pulse-energy femtosecond lasers and corresponding modifications in 2PM setups to accommodate a novel excitation regime. As a direct result of these advancements, 3PM imaging depth in rodent brain is greatly extended with respect to 2PM imaging depth—from ~700 μm to more than 1,200 μm. Due to such a large improvement, we anticipate that 3PM will allow breakthrough studies of the structure (e.g., vascular, glial, and neural architecture) and function (e.g., oxygenation, calcium signaling, and cellular interactions) in preclinical in vivo studies, with cellular- size resolution in previously inaccessible tissue layers, such as the subcortical white matter and the hippocampus. The 11 NIH-funded projects listed in this proposal will significantly benefit from these imaging capabilities. These projects span a diverse range of research topics relevant to NIMH mission, including improvement of the quantitativeness of the clinical imaging modalities and better understanding of the mechanisms involved in development of various brain conditions. The Martinos Center for Biomedical Imaging and Massachusetts General Hospital constitute a diverse community of researchers, many of whom are funded with NIH support, performing cutting edge biomedical research that will further advance our fundamental understanding of human health and disease. This proposed 3PM imaging system will represent the first instrument of its kind in the Boston area (and probably Northeast US) that is available within an Imaging Core. This cutting-edge instrumentation will allow our research community to significantly enhance various research studies relevant to NIMH mission, and it will also allow the Martinos Center to continue prospering as a leader for advanced biomedical imaging.