Mass Spectrometry based molecular imaging of native biological nanodomains

标题：《基于质谱学的天然生物纳米结构域成像》 6.项目摘要/摘要 这份申请为博士后弗朗西斯科·A·费尔南德斯-利马博士提供了职业发展建议 受过生物质谱学和仪器方法开发方面的培训。费尔南德斯博士- 利马致力于生物物理科学的研究生涯，在一个 通过创造和改进当前的技术和方法，在分子和细胞层面上，可以 进一步转化为以动物为基础的研究，以患者为导向的临床研究，并最终导致 改善病人护理。申请者将由Emile A.Schweikert博士在纳米尺度上指导 用于质谱学的成像探针，大卫·H·罗素博士在《仪器和方法》一书中与人共同指导 生物质谱学的发展，由Jennifer L.Bizon博士在行为学中共同指导 以及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农工大学进行，提出了一种工具开发 与可询问原生生物的纳米成像探头耦合的质谱仪 单细胞和亚细胞级别的表面(目前不适用于建议的级别)。这个 仪器(特定目标1)将使用高达100 qkeV的集束探头(Au100n+Q和Binq+Q) 增强的分子产额发射的能量(~10倍增加)，以及分子离子局域化 使用电子发射显微镜实现100 nm以下的横向分辨率。方法论是 使用包含已知表面标记的明确定义的细胞系统验证(例如，表达 来自免疫细胞的CD4抗原和肝细胞生长因子受体(c-met)(Molt-3)和 肝细胞)，以表征仪器性能(特定目标2)。快速气相分离 (在这种情况下，离子迁移率-质谱仪，IM-MS)和碎裂技术(IM-CID-MS)将 应用于分子生物标记物的分离和鉴定(具体目标3)。作为一篇短文- 学期目标，神经元的表型表达、形态和/或稳定性将与 青壮年行为特征的基底前脑化学环境 F344只大鼠(特定目标4)。相关性：该项目将建立工具和方法基础 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 通过直接将化学环境与其生物功能相关联的疾病使用未经处理的 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.