Highly multiplexed ion-beam tissue molecular imaging with sub-micron resolution

具有亚微米分辨率的高度多重离子束组织分子成像

• 批准号: 8664235
• 负责人: GARRY P NOLAN
• 金额: $ 41.06万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8664235
• 关键词: Address; Antibodies; Antigens; Basic Science; Binding; Biological; Biological Testing; Biology; Cell Nucleus; Cell division; Cells; Clinical; Computer software; Data; Detection; Epitopes; Equipment; Evaluation; Formalin; Goals; Histology; Image; Immune system; Immunohistochemistry; Ions; Label; Laser Scanning Confocal Microscopy; Lasers; Light; Location; Malignant Neoplasms; Metals; Methods; Metric; Microscopic; Molecular; Molecular Target; Monitor; Morphology; Neoplasm Metastasis; Noise; Nuclear Proteins; Organelles; Oxygen; Paraffin Embedding; Pathway interactions; Performance; Pharmaceutical Preparations; Phenotype; Pre-Clinical Model; Preparation; Proteins; Reproducibility; Research; Resolution; Retrieval; Sampling; Scanning; Science; Signal Pathway; Signal Transduction; Techniques; Technology; Time; Tissue Sample; Tissues; Translational Research; Validation; Weight; base; cancer therapy; detector; drug development; fluorophore; high standard; imaging Segmentation; imaging modality; insight; instrumentation; malignant breast neoplasm; molecular imaging; protein distribution; prototype; public health relevance; sample fixation; submicron; success; tissue preparation; tool; tumor

DESCRIPTION (provided by applicant): The ability to visualize the presence, abundance, location and functional state of many proteins at once in cells and tissues with preserved morphology has long been a goal. This proposal seeks to optimize recently demonstrated mass-spec-based imaging methods that can, when fully developed, detect as many as 40 to 100+ molecular targets in cells and tissues using simultaneously applied mass-tag-labeled antibodies, with outstanding morphology. The technique is analogous to laser-scanning confocal microscopy, except that the scanning is performed using a tightly focused ion beam, rather than a similarly sized point of laser light. And instead of fluorophores, the labels detected are clustes of metal atoms attached to any given probe. Preliminary data using a highly focused, scanning oxygen ion beam and 10 antibodies applied simultaneously to clinical breast cancer tissues demonstrate histology-like images that reveal bound antibody location with subcellular resolution. Going forward, we would like to optimize sensitivity and reliability, with the goal of making this a standard, high-performance, imaging approach for basic and translational science discovery, drug development, and possible clinical deployment. The present project will address: reliable pre-analytical sample preparation methods and labeling techniques for protein analytes; ion-beam imaging method improvements based on current and projected instrumentation; powerful, easy-to-use software for equipment control, display and analysis; and validation in several important biological test cases.

描述（由申请人提供）：在具有保留形态的细胞和组织中同时可视化许多蛋白质的存在、丰度、位置和功能状态的能力一直是一个目标。 该提案旨在优化最近证明的基于质谱的成像方法，当完全开发时，可以使用同时应用的质量标签标记的抗体检测细胞和组织中多达40至100+个分子靶标，具有出色的形态学。该技术类似于激光扫描共聚焦显微镜，除了使用紧密聚焦的离子束进行扫描，而不是类似大小的激光点。检测到的标签不是荧光团，而是附着在任何给定探针上的金属原子簇。使用高度聚焦的扫描氧离子束和同时应用于临床乳腺癌组织的10种抗体的初步数据展示了具有亚细胞分辨率的显示结合抗体位置的组织学样图像。展望未来，我们希望优化灵敏度和可靠性，目标是使其成为基础和转化科学发现、药物开发和可能的临床部署的标准、高性能成像方法。本项目将涉及：可靠的分析前样品制备方法和蛋白质分析物的标记技术;在现有和预计的仪器基础上改进离子束成像方法;用于设备控制、显示和分析的功能强大、易于使用的软件;以及在几个重要生物测试案例中的验证。