High Resolution Fluorescence Imaging of Dense Brain Regions with Dual Selective Plane Illumination Microscopy

项目摘要/摘要 我们正在申请资金，以支持购买ASI双选择平面照明显微镜 透明组织(ct-dSPIM)显微镜系统。这台仪器将被安置在神经科学 北卡罗来纳大学医学院的显微镜核心(NMC)。这一核心的任务是 设施是为固定和活体的细胞和分子成像提供全光谱的先进系统 样本，以实施新的成像技术，并提供培训、咨询、数据分析、图像 处理和集中技术专门知识，以支持北卡罗来纳大学和其他研究人员的成像需求。 高分辨率成像以区分大脑密集区域中的细胞是必不可少的 中心提供的服务。NMC目前不支持光片显微镜。多重 使用NMC样例进行的竞争演示已确定所请求的ct-dSPIM系统最适合 以高分辨率对iDISCO清除的小鼠脑组织进行成像。所请求的系统是对 当前的LaVision光片显微镜，目前受到较低分辨率硬件功能的限制(请参见 阿里尔博士的支持信)。 NIMH支持的多个不同项目在本申请中展示了对该仪器的需求 在神经生物学、遗传学、病理学、心理学、细胞生物学、发育生物学和 癌症生物学。我们描述了NIMH资助的10个项目，它们需要使用高分辨率XY和光学 由ASI ct-dSPIM提供的切片，以便以高空间分辨率区分精细的细胞特征 在大的密集堆积的体积内(例如，在整个成年小鼠大脑中，密集地 小鼠胚胎大脑的填充区域，以及用于对大的人脑组织进行详细成像)，并需要 在原始的完整3D环境中对大组织进行成像。 NMC自18年前开始运作以来，已成为医学研究不可或缺的一部分 目前支持60多个由美国国立卫生研究院资助的研究项目。收购ASI双选择 用于透明组织的平面照明显微镜(ct-dSPIM)显微镜系统是 NMIH资助的调查人员可以使用的仪器装备，以便不仅保持当前的 为NIMH资助的项目提供高水平的关键支持，但也通过提供现代化的 成像能力。

Project Summary/Abstract We are requesting funds to support the purchase of an ASI Dual Selective Plane Illumination Microcopy for Cleared Tissue (ct-dSPIM) Microscope System. This instrument will be housed within the Neuroscience Microscopy Core (NMC) at the University of North Carolina (UNC) School of Medicine. The mandate of this core facility is to provide a full spectrum of advanced systems for cellular and molecular imaging of fixed and live samples, to implement new imaging technologies, and to offer training, consultation, data analysis, image processing, and centralized technical expertise to support the imaging needs of UNC and other researchers. High-resolution imaging to discriminate between cells in closely-packed regions of the brain is an essential service provided by the center. The NMC currently does not support a light-sheet microscope. Multiple competitive demos with NMC samples have determined that the requested ct-dSPIM system is best suited for imaging iDISCO-cleared mouse brain tissue at high-resolution. The requested system is complementary to the current LaVision Lightsheet microscope, which is currently limited by lower-resolution hardware capabilities (see Letter of Support from Dr. Ariel). The need for this instrument is demonstrated in this application with multiple diverse NIMH supported projects in fields including; Neurobiology, Genetics, Pathology, Psychology, Cell Biology, Developmental Biology, and Cancer Biology. We describe 10 NIMH-funded projects that require the use of high-resolution xy and optical sectioning provided by the ASI ct-dSPIM, in order to distinguish fine cellular features with high spatial resolution within large densely packed volumes (e.g., extended circuitry throughout an adult mouse brain, in densely packed regions of a mouse embryonic brain, and for detailed imaging of large human brain tissue) and require imaging of large tissues in their original full 3D context. Since the NMC commenced operations 18 years ago, it has become an integral part of the medical research community, currently supporting over 60 NIH-funded research projects. Acquisition of an ASI Dual Selective Plane Illumination Microcopy for Cleared Tissue (ct-dSPIM) Microscope System is an essential addition to the repertoire of instrumentation available to NMIH-funded investigators in order to not only maintain the current high-level of critical support for NIMH-funded projects, but also to enhance their research by providing modern imaging capabilities.