Mass Spectrometry based molecular imaging of native biological nanodomains

标题：“基于质谱的天然生物纳米结构域成像” 6.项目总结/摘要 申请书提出了弗朗西斯科A博士的职业发展。Fernando-Lima，博士后 在生物质谱和仪器及方法开发方面接受过培训的研究员。费尔南德斯医生- 利马致力于生物物理科学的研究事业，以研究科学问题， 分子和细胞水平，通过创造新的和改进现有的技术和方法， 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 改善病人护理。申请人将由Emile A博士指导。纳米尺度的Schweikert 质谱成像探针，由大卫H.罗素在仪器和方法 生物质谱的发展，并共同指导博士詹妮弗L。行为学中的比松 以及用于研究认知障碍疾病的细胞神经科学方法和动物模型。 该项目将在得克萨斯A&M大学进行，提出了一个工具的发展， 与纳米成像探针耦合的质谱仪， 在单细胞和亚细胞水平上的表面（目前在提议的水平上不可用）。的 仪器（Specific Aim 1）将采用高达100 qkeV的集束束探针（Au 100 n +q和Binq+q 增强的分子产率发射（~10倍增加）和分子离子定位的能量， 使用电子发射显微镜，横向分辨率低于100 nm。方法将是 使用含有已知表面标记的明确的细胞系统（例如，表达 来自免疫细胞（Molt-3）的CD 4抗原和肝细胞生长因子受体（c-met），以及 肝细胞），以表征仪器性能（具体目标2）。快速气相分离 (in在这种情况下，离子迁移-质谱法（IM-MS）和碎裂技术（IM-CID-MS）将 应用于分子生物标志物的分离和鉴定（具体目标3）。作为一个简短的- 术语目标，神经元表型表达、形态学和/或稳定性将与神经元表型表达、形态学和/或稳定性相关。 青年、中年和老年人行为特征的基底前脑化学环境 F344大鼠（具体目标4）。相关性：该项目将奠定工具和方法基础 用于与认知障碍相关的分子标志物的单细胞和亚细胞研究 疾病的化学环境与其生物功能直接相关，使用未经处理的 组织样本

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.