Mass Spectrometry based molecular imaging of native biological nanodomains

基于质谱的天然生物纳米域分子成像

• 批准号: 8515552
• 负责人: Francisco Fernandez-Lima
• 金额: $ 24.87万
• 依托单位: FLORIDA INTERNATIONAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8515552
• 关键词: Acceleration; Age; Animal Model; Animals; Behavioral; Biological; Biological Markers; Biological Markers; Biological Process; CD4 Antigens; Calibration; Cells; Chemicals; Chemistry; Clinical Research; Commit; Coupled; Detection; Development; Device or Instrument Development; Disease; Electrons; Environment; Gases; Generations; Goals; Hepatocyte; Image; Immune; Impaired cognition; Inbred F344 Rats; Ions; Lasers; Lateral; Lead; Mass Spectrum Analysis; Mediating; Mentors; Methodology; Methods; Microscope; Molecular; Molting; Morphology; Neurological Models; Neurons; Neurosciences; Patient Care; Pattern; Performance; Phase; Postdoctoral Fellow; Procedures; Proto-Oncogene Protein c-met; Research; Resolution; Sampling; Science; Signal Transduction; Staging; Surface; System; Techniques; Technology; Texas; Tissue Sample; Tissues; Training; Translating; Universities; Work; aged; basal forebrain; base; career; career development; cholinergic; design; imaging probe; improved; instrument; interest; ion mobility; ionization; light microscopy; mass spectrometer; meetings; method development; middle age; molecular imaging; molecular marker; nanometer; nanoscale; novel; patient oriented; tool

Title: "Mass Spectrometry based imaging of native biological nanodomains" 6. PROJECT SUMMARY/ABSTRACT The application proposes a career development for Dr. Francisco A. Fernandez-Lima, a postdoctoral fellow trained in biological mass spectrometry and instrument & method development. Dr. Fernandez- Lima is committed to a research career in biophysical science to study scientific problems at a molecular and cellular levels by creating new and improving current techniques and methods, that can be further translated to animal based studies, to patient-oriented clinical research, and ultimately lead to improved patient care. The applicant will be mentored by Dr. Emile A. Schweikert in nanometer scale imaging probes for mass spectrometry, co-mentored by Dr. David H. Russell in instrument and method development for biological mass spectrometry, and co-mentored by Dr. Jennifer L. Bizon in behavioral and cellular neuroscience methods and animal models for studies of cognitive impairment diseases. The project, to be conducted at Texas A&M University, proposes the instrumental development of a mass spectrometer coupled to a nanometer imaging probe capable of interrogating native biological surfaces at the single cell and sub-cellular levels (currently not available at the level proposed). The instrument (Specific Aim 1) will employ a cluster beam probe (Au100n+q and Binq+q) at up to 100 qkeV energies for enhanced molecular yield emission (~10 fold increase), and molecular ion localization with sub-100nm lateral resolution using an electron emission microscope. The methodology will be validated using well-defined cellular systems containing known surface markers (e.g., expression of CD4 antigen and hepatocyte growth factor receptor (c-met) from Immune cells (Molt-3) and hepatocytes) to characterize the instrument performance (Specific Aim 2). Fast gas-phase separation (in this case Ion Mobility - Mass Spectrometry, IM-MS) and fragmentation techniques (IM-CID-MS) will be applied to the separation and identification of molecular biomarkers (Specific Aim 3). As a short- term goal, the neuron phenotypic expression, morphology, and/or stability will be correlated with the basal forebrain chemical environment of behaviorally characterized young, middle-aged, and aged F344 rats (Specific Aim 4). Relevance: The project will set the instrumental and methodological basis for single cell and sub-cellular studies of molecular markers associated with cognitive impairment diseases by directly correlating the chemical environment with their biological function using untreated tissue samples.

标题：“基于质谱的天然生物纳米域的成像” 6。项目摘要/摘要 该申请提出了弗朗西斯科·A·费尔南德斯·利马博士的职业发展，博士后 接受过生物质谱和仪器和方法开发的研究员。费尔南德斯博士 利马致力于生物物理科学研究生涯，以研究 分子和细胞水平通过创建新的和改进的当前技术和方法，可以 进一步转化为基于动物的研究，以患者为导向的临床研究，并最终导致 改善患者护理。申请人将由Emile A. Schweikert博士以纳米量表进行指导 David H. Russell博士在仪器和方法中联合征收质谱的成像探针 生物学质谱的开发，并由Jennifer L. Bizon博士合作。 以及用于研究认知障碍疾病研究的细胞神经科学方法和动物模型。 该项目将在德克萨斯A＆M大学进行，提出了 质谱仪耦合到能够询问天然生物学的纳米成像探针 单细胞和亚细胞水平的表面（目前在建议的级别上都不可用）。这 仪器（特定目标1）将在多达100 QKEV处采用簇梁探针（AU100N+Q和BINQ+Q） 增强分子屈服发射的能量（增加了10倍），并与分子离子定位 使用电子发射显微镜的横向分辨率低于100nm。该方法将是 使用含有已知表面标记的明确定义的蜂窝系统验证（例如，表达 来自免疫细胞（MOLT-3）的CD4抗原和肝细胞生长因子受体（C-MET）和 肝细胞表征仪器性能（特定目标2）。快速气相分离 （在这种情况下，离子迁移率 - 质谱，IM-MS）和碎片技术（IM-CID-MS）将 应用于分子生物标志物的分离和鉴定（特定目标3）。作为一个简短的 术语目标，神经元表型表达，形态和/或稳定性将与 行为表征的年轻，中年和老年的基础前脑化学环境 F344大鼠（特定目标4）。相关性：该项目将设定工具和方法论基础 用于与认知障碍相关的分子标记的单细胞和亚细胞研究 通过使用未处理的化学环境将化学环境与其生物功能直接相关的疾病 组织样品。