Mass Spectrometry based molecular imaging of native biological nanodomains

描述（由申请人提供）：该申请提出了弗朗西斯科A博士的职业发展。Fernando-Lima博士是一名博士后研究员，接受过生物质谱和仪器与方法开发方面的培训。费尔南德斯-利马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，并最终改善患者护理。申请人将由Emile A博士指导。Schweikert在纳米尺度成像探针质谱，共同指导博士大卫H。罗素在生物质谱仪的仪器和方法开发，并共同指导博士詹妮弗L。行为和细胞神经科学方法和认知障碍疾病研究的动物模型。该项目将在得克萨斯A&M大学进行，提出了一种质谱仪的仪器开发，该质谱仪与纳米成像探针耦合，能够在单细胞和亚细胞水平上询问天然生物表面（目前在拟议的水平上不可用）。该仪器（具体目标1）将采用能量高达100 qkeV的簇束探针（Au 100 n +q和Binq+q），以增强分子产额发射（约增加10倍），并使用电子发射显微镜以低于100 nm的横向分辨率进行分子离子定位。该方法将使用含有已知表面标志物（例如，免疫细胞（Molt-3）和肝细胞的CD 4抗原和肝细胞生长因子受体（c-met）表达），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-质谱法，IM-MS）和裂解技术（IM-CID-MS）将用于分离和鉴定分子生物标志物（具体目标3）。作为短期目标，神经元表型表达、形态和/或稳定性将与行为表征的年轻、中年和老年F344大鼠的基底前脑化学环境相关（具体目标4）。相关性：该项目将通过使用未经处理的组织样本将化学环境与其生物功能直接联系起来，为与认知障碍疾病相关的分子标志物的单细胞和亚细胞研究奠定仪器和方法学基础。 公共卫生相关性：细胞水平的生物学性能由化学环境和表面化学介导。将创建一种新的仪器和方法，可以检查天然生物表面上的分子组成。一个独特的功能将是生物标记的本地化，其分辨率比光学显微镜提高了一百倍。

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.