Development of VSSI-probe technology for in situ probing biological systems using mass spectrometry

开发使用质谱法原位探测生物系统的 VSSI 探针技术

• 批准号: 10468735
• 负责人: Peng Li
• 金额: $ 33.9万
• 依托单位: WEST VIRGINIA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-23 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10468735
• 关键词: Address; Adopted; Adoption; Biological; Biological Assay; Biological Process; Biomedical Research; Cell Culture Techniques; Cells; Chemicals; Chemistry; Communities; Complex; Detection; Development; Diagnosis; Diagnostic; Environment; Enzymatic Biochemistry; Frequencies; Gases; Geometry; Glass; Human; Huntington Disease; Image; Immobilized Enzymes; In Situ; Ions; Kinetics; Knowledge; Lasers; Lead; Lipid Bilayers; Lipids; Liquid substance; Maps; Mass Spectrum Analysis; Mechanics; Methods; Microbe; Microscope; Monitor; Nebulizer; Organism; Oxidative Stress; Performance; Phase; Preparation; Procedures; Proteins; Proteomics; Reaction; Research; Resolution; Sampling; Slice; Slide; Solid; Solvents; Source; Spectrum Analysis; Speed; Surface; System; Techniques; Technology; Time; Tissue Sample; Tissues; Transducers; Work; base; biological systems; biomaterial compatibility; clinical diagnostics; cost; design; detection sensitivity; detector; direct application; experimental study; flexibility; high sensitivity probe; imaging platform; improved; in vivo; instrument; ionization; ionization technique; lipidomics; mass analyzer; mass spectrometric imaging; metabolomics; monolayer; operation; protein aggregation; structural biology; temporal measurement; tool; voltage

Project Summary Mass spectrometry (MS) is a powerful tool for biological studies, which has found a wide range of applications including proteomics, metabolomics, glycomics, lipidomics, and structural biology. While most of the existing MS applications are extraction-based batch analysis, using MS for in situ or in vivo analysis of biological samples has become increasingly important to enable new biological discoveries. The past decade has witnessed the rapid development of interfacing methods for direct MS analysis of various biological samples from tissue slices to single cells. However, existing methods still have difficulties in studying living and dynamic biological systems in a non-invasive manner and in situ MS experiments are primarily performed by MS research groups due to the involvement of complex instrument setups. As a result, these limitations significantly hinder the progress of using direct MS for biological discoveries. To address the unmet need for in situ MS analysis of biological systems, our objective is to develop a direct MS method, vibrating sharp-edge spray ionization probe (VSSI-probe), that is simple, easy to use, biocompatible, and amenable to high speed sampling. Compared to existing methods, the proposed VSSI-probe is amenable to fast sampling, highly biocompatible, voltage-free, easy to operate, and substantially less expensive. Therefore, we expect that once demonstrated, the proposed VSSI-probe will enable new MS applications as well as improve the adoption of direct MS analysis in the biological community. In particular, we will (1) optimize the performance of the VSSI-probe for high sensitivity MS detection;(2) demonstrate a high-throughput VSSI-probe platform for studying fast biological reactions;(3) demonstrate the utility of VSSI-probe in studying a delicate and dynamic biological system of lipid/protein aggregates interaction. (4) demonstrate the high spatial resolution sampling capability of VSSI-probe for studying metabolites directly from bacterial colony. With the unprecedented capability and flexibility of probing biological samples, the proposed VSSI-probe will enable a broad range of biological applications that requires fast and biocompatible sampling including enzymology, cell monitoring, lipid bilayer chemistry, and real time intraoperative diagnosis. The simplicity, low cost, and compatibility with existing mass analyzers of the proposed system will also greatly reduce the technical barrier for adopting this technology by non-MS groups thereby expanding applications of direct MS to more diverse biological problems.

项目摘要 质谱学是生物学研究的有力工具，有着广泛的应用前景 包括蛋白质组学、代谢组学、糖组学、脂组学和结构生物学。而现有的大多数MS 应用是基于萃取的批处理分析，使用MS对生物样品进行原位或活体分析 对于实现新的生物发现变得越来越重要。过去的十年见证了 直接从组织切片中分析各种生物样品的接口方法的快速发展 到单个细胞。然而，现有的方法在研究生命和动态生物系统方面仍然存在困难。 以非侵入性方式和现场MS实验主要由MS研究小组执行，因为 涉及复杂的仪器设置。因此，这些限制大大阻碍了使用 用于生物发现的直接MS。为了满足对生物系统的原位MS分析的未满足的需求， 我们的目标是发展一种直接的MS方法，振动尖端喷雾电离探头(VSSI-Probe)， 简单，易于使用，生物兼容性，并服从于高速采样。与现有方法相比， 所提出的VSSI探头具有快速采样、高度生物兼容性、无电压、易于操作和 大大降低了成本。因此，我们预计，一旦展示，拟议的VSSI-Probe将使 新的MS应用，以及促进在生物界采用直接MS分析。在……里面 具体地说，我们将(1)优化VSSI-探测器的性能，以实现高灵敏度的MS检测；(2) 展示用于研究快速生物反应的高通量VSSI-Probe平台；(3)展示 VSSI-Probe在研究微妙而动态的脂质/蛋白质聚集体相互作用生物系统中的应用。 (4)展示了VSSI-Probe用于直接研究代谢产物的高空间分辨率采样能力。 来自细菌菌落。凭借前所未有的探测生物样本的能力和灵活性， 建议的VSSI-Probe将使需要快速和生物兼容的广泛生物应用成为可能 采样包括酶学、细胞监测、脂质双层化学和术中实时诊断。 该系统的简单性、低成本和与现有质量分析器的兼容性也将大大 减少非微软团队采用这项技术的技术障碍，从而扩大 引导多发性硬化症解决更多的生物问题。