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: 10021677
• 负责人: Peng Li
• 金额: $ 34.2万
• 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/10021677
• 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; detector; direct application; experimental study; flexibility; high sensitivity probe; imaging platform; improved; in vivo; instrument; ionization; ionization technique; mass analyzer; 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（MS）是生物学研究的强大工具，该工具已发现了广泛的应用 包括蛋白质组学，代谢组学，糖基因，脂肪组学和结构生物学。而大多数现有的MS 应用是基于提取的批处理分析，使用MS进行原位或体内生物样品分析 实现新的生物学发现变得越来越重要。过去十年见证了 快速开发用于直接MS分析组织切片的各种生物样品的接口方法 到单个细胞。但是，现有方法在研究生活和动态生物系统方面仍然存在困难 MS研究小组主要以非侵入性和原位MS实验进行 复杂仪器设置的参与。结果，这些限制显着阻碍了使用的进度 直接用于生物学发现的MS。为了满足对生物系统原位MS分析的未满足的需求， 我们的目标是开发直接的MS方法，振动尖锐的喷雾电离探针（VSSI-Probe），即 简单，易于使用，生物相容性且可适应高速采样。与现有方法相比 拟议的VSSI-Probe可以随时采样，高度生物相容性，无电压，易于操作，并且 价格较低。因此，我们希望一旦证明，拟议的VSSI-Probe将启用 新的MS应用程序，并改善了生物群落中直接MS分析的采用。在 特别是，我们将（1）优化对高灵敏度MS检测的VSSI-probe的性能；（2） 展示了一个用于研究快速生物学反应的高通量VSSI-probe平台；（3）证明了该平台 VSS-probe在研究脂质/蛋白质聚集体相互作用的微妙而动态的生物系统方面的效用。 （4）证明了直接研究代谢物的VSSI-probe的高空间分辨率采样能力 来自细菌菌落。具有前所未有的能力和探测生物样品的灵活性， 拟议的VSSI-Probe将实现广泛的生物应用，需要快速和生物相容性 采样包括酶学，细胞监测，脂质双层化学和实时术中诊断。 与拟议系统的现有大量分析仪的简单性，低成本和兼容性也将大大 减少非MS小组采用这项技术的技术障碍，从而扩大了 将MS引向更多样化的生物学问题。