Spatial Analysis of Molecules in Tissue using MALDI-MS

使用 MALDI-MS 对组织中的分子进行空间分析

• 批准号: 7630527
• 负责人: RICHARD M CAPRIOLI
• 金额: $ 44.48万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7630527
• 关键词: 3-Dimensional; Address; Animal Organ; Animals; Area; Biological; Brain; Chemistry; Detection; Detergents; Development; Digestion; Disease; Goals; Health; Histology; Human; Image; Image Enhancement; In Situ; Ions; Light; Maintenance; Maps; Mass Spectrum Analysis; Measures; Metabolism; Methodology; Methods; Molecular; Molecular Probes; Mus; Neurobiology; Organ; Parkinsonian Disorders; Pattern; Peptide Hydrolases; Peptides; Performance; Pharmaceutical Preparations; Preparation; Prevention; Process; Protein Analysis; Protein Array; Protein Family; Proteins; Protocols documentation; Rattus; Reagent; Reproducibility; Reproductive Biology; Research; Resolution; Sampling; Signal Transduction; Solubility; Space Perception; Spatial Distribution; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Speed; Structure; Technology; Testing; Tissues; Toxic effect; Validation; Work; base; biological research; drug distribution; human disease; improved; insight; instrumentation; mouse model; natural Blastocyst Implantation; new technology; next generation; particle; submicron

DESCRIPTION (provided by applicant): The broad range goals of the proposed research involves development of MALDI Imaging Mass Spectrometry (IMS) to achieve (i) next generation improvements of the technology and associated methodologies for both profiling and imaging of tissues to gain significant improvements in speed, sensitivity and image resolution, (ii) development, testing, and validation of in situ chemistries for targeted analysis of specific proteins or families of proteins at high sensitivity, including in situ protein identification and (iii) applications of IMS to several biological research problems that can significantly benefit from this technology, e.g., addressing several fundamental questions in neurobiology, reproductive biology (embryo implantation), and drug distribution, toxicity and efficacy in intact organs and animal sections. More specifically, work proposed for technology and methodology improvements involve next generation enhancements for image acquisition speed, image resolution, and ease, robustness, and reproducibility of image acquisition. New sample preparation protocols will be devised to facilitate histology-directed molecular tissue analysis and to provide ease of 3-D protein image construction of tissue structures and small organs. Targeted analysis of proteins in tissues using in situ chemistries (tissue-based protein arrays) will be developed for the identification and quantitation of proteins directly in tissue sections. These include protocols that allow protease digestions in micron sized areas in tissue and the use of submicron particles having light cleavable linkers attached to molecular probes to remove and analyze proteins or families of proteins at very high sensitivity with spatially fidelity to the original tissue. Finally, these technologies and protocols will be applied to specific research problems to establish their efficacy in helping solve biological problems and provide spatially resolved molecular information. These applications will include problems in neurobiology, including studies in a mouse model of Parkinsonism, the creation of 3-D maps of mouse brain proteins, studies of embryo implantation in mouse, and applications to drug analysis, metabolism, and efficacy in studies in rat and mouse through tissue, organ, and whole body analyses. This research will provide a new technology for the discovery of molecular patterns in both health and disease. The elucidation of the spatial orientation of proteins in tissues and their quantitation and specific identification will answer many scientific questions about how disease might form and spread and will bring new insights into the prevention and treatment of human disease and the maintenance of human health.

描述（由申请人提供）：拟议的研究的广泛范围涉及发展MALDI成像质谱法（IMS），以实现（i）（i）技术和相关方法的下一代改进，用于分析组织的分析和成像，从IMS在几个生物学研究问题上的原位蛋白质鉴定和（iii）应用，这些问题可以从该技术中显着受益，例如，解决了神经生物学，生殖生物学（胚胎植入）以及药物分布，毒性和效率在完整的器官和动物区域中的几个基本问​​题。更具体地说，针对技术和方法改进提出的工作涉及图像采集速度，图像分辨率以及轻松，鲁棒性以及图像获取的可重复性的下一代增强功能。将设计新的样品制备方案，以促进组织学指导的分子组织分析，并提供3-D蛋白质图像的易于组织结构和小型器官。使用原位化学（基于组织的蛋白阵列）对组织中蛋白质的有针对性分析将直接在组织切片中直接鉴定和定量蛋白质。这些方案包括允许在组织中的微米大小区域消化的方案，以及使用与分子探针连接到的光线可裂解接头的亚微米颗粒，以去除和分析蛋白质或蛋白质的蛋白质或蛋白质家族，以非常高的敏感性，并在空间上延伸到原始组织。最后，这些技术和方案将应用于特定的研究问题，以确立其在帮助解决生物学问题并提供空间解决的分子信息方面的功效。这些应用将包括神经生物学中的问题，包括在帕金森氏症小鼠模型中的研究，小鼠脑蛋白的3-D地图，小鼠中胚胎植入的研究以及在药物分析，代谢和通过组织，器官和整个身体分析中在大鼠和小鼠中的研究中的应用。这项研究将为发现健康和疾病的分子模式提供新技术。阐明组织中蛋白质的空间取向及其定量和特定识别将回答有关疾病如何形成和传播的许多科学问题，并将为预防和治疗人类疾病以及维持人类健康带来新的见解。