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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

基本信息

批准号：
10687118
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
项目类别：
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/10687118
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 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 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 vibration voltage

项目摘要

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.

项目概要质谱 (MS) 是生物学研究的强大工具，有着广泛的应用包括蛋白质组学、代谢组学、糖组学、脂质组学和结构生物学。虽然大多数现有的 MS 应用是基于提取的批量分析，使用 MS 对生物样品进行原位或体内分析对于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对组织切片中的各种生物样品进行直接 MS 分析的接口方法到单细胞。然而，现有方法在研究活体和动态生物系统方面仍然存在困难以非侵入性方式进行的原位 MS 实验主要由 MS 研究小组进行，因为涉及复杂的仪器设置。因此，这些限制极大地阻碍了使用的进展直接 MS 用于生物学发现。为了解决生物系统原位 MS 分析未得到满足的需求，我们的目标是开发一种直接 MS 方法，即振动锐边喷雾电离探针（VSSI 探针），简单、易于使用、生物相容且适合高速采样。与现有方法相比，所提出的 VSSI 探针适合快速采样、高度生物相容性、无电压、易于操作，并且便宜得多。因此，我们预计，一旦得到证实，所提出的 VSSI 探针将能够新的 MS 应用，并提高直接 MS 分析在生物界的采用。在特别是，我们将 (1) 优化 VSSI 探针的性能，以实现高灵敏度 MS 检测；(2) 展示用于研究快速生物反应的高通量 VSSI 探针平台；(3) 展示 VSSI 探针在研究脂质/蛋白质聚集体相互作用的微妙且动态的生物系统中的效用。 (4) 展示 VSSI 探针用于直接研究代谢物的高空间分辨率采样能力来自细菌菌落。凭借探测生物样品的前所未有的能力和灵活性，拟议的 VSSI 探针将实现需要快速且生物相容性的广泛生物应用采样包括酶学、细胞监测、脂质双层化学和实时术中诊断。该系统的简单性、低成本以及与现有质量分析仪的兼容性也将大大提高降低非 MS 群体采用该技术的技术障碍，从而扩大该技术的应用将 MS 引导至更多样化的生物学问题。