Multiparametric deep tissue microscope for in vivo and in vitro imaging

用于体内和体外成像的多参数深层组织显微镜

• 批准号: 10426767
• 负责人: Lucy Erin O'brien
• 金额: $ 60万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10426767
• 关键词: 12 year old; Address; Alveolar; Animal Model; Area; Basal Cell; Biological; Biomedical Research; Biophysics; Biotechnology; Bone Marrow; Cells; Communities; Computers and Advanced Instrumentation; Confocal Microscopy; Detection; Development; Disease; Electron Microscopy Facility; Equipment; Explosion; Funding; Health; Hereditary Disease; Human; Image; In Vitro; Individual; Laboratory Research; Lung; Methods; Microscope; Microscopic; Microscopy; Molecular; Osteogenesis; Photons; Procedures; Proteins; Research; Research Project Grants; Research Support; Resource Sharing; Resources; Sampling; Scanning; Science; Signal Transduction; Skin Cancer; Slice; Solid Neoplasm; Specimen; Stains; Techniques; Thick; Thinness; Tissues; Touch sensation; United States National Institutes of Health; Universities; Visualization; ex vivo imaging; gene therapy; high resolution imaging; imaging facilities; imaging system; in vivo; instrument; light microscopy; microscopic imaging; multi-photon; multiphoton microscopy; optical imaging; programs; second harmonic generation imaging; skeletal stem cell; stem cell niche; stem cells; tissue culture; tumor

Project Summary: Every cell and tissue biomedical research program requires microscopic visualization of research specimens. Some of this need can be addressed by routine widefield and confocal microscopy of relatively thin (< 50 um) tissue slices or tissue culture. However, biological understanding of molecular, cell and tissue-level dynamics has been uniquely enabled by the recent explosion of advanced, multi-parametric microscopy procedures for in vivo and ex vivo imaging. These procedures include multiphoton microscopy combined with harmonically generated signals (SHG) for imaging in thick (100s of microns) tissue and solid tumors, expressing multiple fluorescent proteins, as well as for deep (>1 mm), high resolution imaging of optically cleared tissues (e.g. Clarity, iDISCO) stained with multiple fluorescent markers. These techniques are now a required part of contemporary biomedical cell and tissue research. To be applied successfully, these methods require a versatile, advanced imaging system that incorporates fast scanning, multi-photon and 1-photon excitation at multiple wavelengths, high sensitivity and spectral selectivity over multiple detection channels, rapid imaging, and environmental control. The demands of these state-of-the-art microscopy methods for both specialized equipment and specialized expertise often outstrip the resources available to individual research laboratories. As such, these needs are best met with a shared instrument housed in a shared resource facility. For these reasons we are requesting funding to replace Stanford University's Cell Sciences Imaging Facility's 12- year-old in vivo multi-photon/confocal microscope (Leica SP5) with the new, state-of-the-art Stellaris 8 DIVE microscope. This combined multi-photon, confocal, in vivo, and ex vivo as well as in vitro imaging microscope will be a shared resource located in a well-established, multi-user light and electron microscopy facility: The Cell Sciences Imaging Facility (http://microscopy.stanford.edu). This facility is accessible to Stanford University's entire research community as well as surrounding biotech companies. The Stellaris 8 DIVE microscope will support research projects from 11 users, 10 of which are NIH funded. Their studies investigate critical functional and structural questions in a variety of model organisms and tissues and cover areas of biomedical research with implications for diverse aspects of human health and disease. These projects include: clonal fate and linage tracing of stem cells and tumor initiating cells, interrogation of the alveolar stem cell niche, molecular and cellular mechanisms of small cell lung and basal cell skin cancers development, characterization of skeletal stem cells in osteogenesis, characterization of the bone marrow microenvironment, gene therapy for inherited diseases and the biophysics of touch as well as characterizing. All these projects critically require advanced instrumentation for imaging live and fixed tissue and cell samples, a need which can only uniquely be met by the requested Leica Stellaris 8 DIVE microscope.

项目概要： 每个细胞和组织生物医学研究项目都需要研究样本的显微可视化。 其中一些需求可以通过相对较薄（< 50 um）的常规宽场和共焦显微镜来解决 组织切片或组织培养。然而，对分子、细胞和组织水平动力学的生物学理解 最近先进的多参数显微镜程序的爆炸性发展使这一独特的技术得以实现 体内和离体成像。这些程序包括多光子显微镜与谐波相结合 生成信号（SHG），用于厚（数百微米）组织和实体瘤中的成像，表达多种 荧光蛋白，以及光学透明组织（例如组织）的深层（>1 mm）高分辨率成像 Clarity，iDISCO）用多种荧光标记染色。这些技术现在已成为 当代生物医学细胞和组织研究。要成功应用这些方法，需要 多功能、先进的成像系统，结合了快速扫描、多光子和单光子激发 多个波长、多个检测通道的高灵敏度和光谱选择性、快速成像、 和环境控制。这些最先进的显微方法对专业人士的要求 设备和专业知识往往超出单个研究实验室可用的资源。 因此，这些需求最好通过共享资源设施中的共享仪器来满足。对于这些 我们请求资金来更换斯坦福大学细胞科学成像设施的 12- 一年前的活体多光子/共焦显微镜 (Leica SP5)，配有最先进的新型 Stellaris 8 潜水显微镜。这结合了多光子、共焦、体内、离体以及体外成像 显微镜将成为位于完善的多用户光学和电子显微镜中的共享资源 设施：细胞科学成像设施 (http://microscopy.stanford.edu)。该设施可供以下人士使用 斯坦福大学的整个研究界以及周边的生物技术公司。群星8号 DIVE 显微镜将支持 11 个用户的研究项目，其中 10 个由 NIH 资助。他们的研究 研究各种模型生物和组织中的关键功能和结构问题并涵盖 对人类健康和疾病的各个方面具有影响的生物医学研究领域。这些 项目包括：干细胞和肿瘤起始细胞的克隆命运和谱系追踪、询问 肺泡干细胞生态位、小细胞肺癌和基底细胞皮肤癌的分子和细胞机制 发育、成骨过程中骨骼干细胞的表征、骨髓的表征 微环境、遗传性疾病的基因治疗以及触摸和表征的生物物理学。 所有这些项目都迫切需要先进的仪器来对活体和固定的组织和细胞样本进行成像， 只有 Leica Stellaris 8 DIVE 显微镜才能满足这一需求。