Mass Spectrometry based molecular imaging of native biological nanodomains

描述（由申请人提供）：本申请为弗朗西斯科·a·费尔南德斯-利马博士提供职业发展机会，他是一名从事生物质谱和仪器与方法开发的博士后研究员。Fernandez- Lima博士致力于生物物理科学的研究事业，通过创造新的和改进现有的技术和方法，在分子和细胞水平上研究科学问题，这些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。申请人将接受Emile A. Schweikert博士的纳米级质谱成像探针指导，David H. Russell博士的生物质谱仪器和方法开发指导，Jennifer L. Bizon博士的行为和细胞神经科学方法以及认知障碍疾病研究的动物模型指导。该项目将由德克萨斯A&M大学进行，提出了一种与纳米成像探针相结合的质谱仪的仪器开发，能够在单细胞和亚细胞水平上询问天然生物表面（目前在提议的水平上还没有可用）。该仪器（Specific Aim 1）将采用高达100 qkeV能量的束束探针（Au100n+q和Binq+q）来增强分子产率发射（约10倍），并使用电子发射显微镜进行亚100nm横向分辨率的分子离子定位。该方法将使用包含已知表面标记的定义良好的细胞系统进行验证（例如，来自免疫细胞（Molt-3）和肝细胞的CD4抗原和肝细胞生长因子受体（c-met）的表达），以表征仪器的性能（特定目标2）。快速气相分离（在这种情况下离子迁移-质谱，IM-MS）和碎片化技术（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）。作为短期目标，神经元表型表达、形态和/或稳定性将与具有行为特征的年轻、中年和老年F344大鼠的基底前脑化学环境相关（Specific Aim 4）。相关性：该项目将通过使用未经处理的组织样本直接将化学环境与其生物学功能联系起来，为与认知障碍疾病相关的分子标记的单细胞和亚细胞研究奠定工具和方法基础。

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: The biological performance at the cellular level is mediated by the chemical environment and surface chemistry. A new instrument and method will be created which can examine molecular composition on native biological surfaces. A unique feature will be the localization of biological markers with a resolution improved one hundred-fold over light microscopy.