Interfacing Droplets with Mass Spectrometry for Single-Cell Analysis

将液滴与质谱连接进行单细胞分析

• 批准号: 8539034
• 负责人: FRANTISEK TURECEK
• 金额: $ 20.8万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8539034
• 关键词: Acceleration; Address; Air; Atmospheric Pressure; Biochemical; Blood capillaries; Cancer Detection; Cell Volumes; Cells; Chemicals; Complex; DNA; Detection; Development; Disease; Gene Expression; Goals; Ion Transport; Ions; Knowledge; Laboratories; Lipids; Liquid substance; Mass Spectrum Analysis; Measurement; Microfluidics; Operating System; Optical Methods; Optics; Organism; Peptides; Physiology; Polymerase Chain Reaction; Positioning Attribute; Proteins; Reaction; Research; Resolution; Sampling; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; System; Techniques; Technology; Vacuum; Work; analytical method; base; biological systems; capillary; chemical reaction; design and construction; experience; improved; instrument; interest; ionization; ionization technique; mass spectrometer; meetings; next generation; optical traps; pressure; public health relevance; response; single cell analysis; single molecule; small molecule

This application addresses RFA-GM-1-1009, Exceptional, Unconventional Research Enabling Knowledge Acceleration (EUREKA)(R01). The basic unit of any biological system is the cell, and malfunctions at the single-cell level can result in devastating diseases. To understand the physiology and functions of any biological system, therefore, it is important to achieve measurements with single-cell resolutions. This proposal aims to employ advanced mass spectrometric technologies which, when appropriately combined with new droplet manipulation techniques, will allow us to meet the tremendous challenge of analyzing the contents of single cells with high sensitivty and information content using mass spectrometry. Specifically, the aim of this project is: Design and construct a mass spectrometer with the appropriate interface for ionizing and analyzing the contents of single cells trapped in liquid droplets. The interface will be a droplet nanolab platform with the optical trapping and positioning of droplets in air, which will be used towards achieving the efficient ionization of single-droplet contents and ion transport for mass spectrometry analysis. Atmospheric pressure chemical ionization (APCI) will be used for the analysis of small-molecule (e.g. metabolites and lipids) contents of single cells. MALDI will be used for the analysis of peptide and protein contents of single cells. The cell is the basic unit of any biological system; to understand the functions and malfunctions of an organism will thus necessitate measurements at the single-cell level. Yet the biochemical analysis of single cells is highly challenging, owing to both the minute amounts of sample available as well as the high complexity of the sample. In this proposed project, we aim to tackle these challenges by pushing the limit of sensitivity of mass spectrometry for analyzing the minute but highly concentrated single-cell derived samples present in droplets. PUBLIC HEALTH RELEVANCE: The cell is the basic unit of any biological system; to understand the functions and malfunctions of an organism will thus necessitate measurements at the single-cell level. Yet the biochemical analysis of single cells is highly challenging, owing to both the minute amounts of sample available as well as the high complexity of the sample. In this proposed project, we aim to tackle these challenges by pushing the limit of sensitivity of mass spectrometry for analyzing the minute but highly concentrated single-cell derived samples present in droplets.

此应用程序针对RFA-GM-1-1009，特殊的、非传统的研究支持知识加速(Eureka)(R01)。任何生物系统的基本单位都是细胞，单细胞水平的故障可能会导致毁灭性的疾病。因此，要了解任何生物系统的生理和功能，实现单细胞分辨率的测量是很重要的。这项建议旨在采用先进的质谱学技术，当这些技术与新的液滴操纵技术适当地结合在一起时，将使我们能够应对利用质谱学分析单个细胞的高灵敏度和信息含量的巨大挑战。具体地说，该项目的目标是：设计和建造一个具有适当接口的质谱仪，用于电离和分析捕获在液滴中的单个细胞的内容物。该界面将是一个液滴NanoLab平台，具有对空气中液滴的光学捕获和定位，将用于实现单个液滴内容物的有效电离和用于质谱分析的离子传输。大气压化学电离(APCI)将用于分析单个细胞的小分子(如代谢物和脂类)含量。MALDI将用于分析单细胞的多肽和蛋白质含量。细胞是任何生物系统的基本单位；因此，要了解生物体的功能和故障，就必须在单细胞水平上进行测量。然而，由于可获得的样本数量很少，而且样本的复杂性很高，单细胞的生化分析具有很大的挑战性。在这个拟议的项目中，我们的目标是通过突破质谱学的灵敏度极限来应对这些挑战，以分析液滴中存在的微小但高度浓缩的单细胞衍生样品。 与公共健康相关：细胞是任何生物系统的基本单位；因此，要了解生物体的功能和故障，就必须在单细胞水平上进行测量。然而，由于可获得的样本数量很少，而且样本的复杂性很高，单细胞的生化分析具有很大的挑战性。在这个拟议的项目中，我们的目标是通过突破质谱学的灵敏度极限来应对这些挑战，以分析液滴中存在的微小但高度浓缩的单细胞衍生样品。

项目成果

Modulating patterns of two-phase flow with electric fields.

用电场调制两相流模式。

• DOI: 10.1063/1.4891099
• 发表时间: 2014
• 期刊: Biomicrofluidics
• 影响因子: 3.2
• 作者: Liu,Dingsheng;Hakimi,Bejan;Volny,Michael;Rolfs,Joelle;Anand,RobbynK;Turecek,Frantisek;Chiu,DanielT
• 通讯作者: Chiu,DanielT

Nanoliter segmented-flow sampling mass spectrometry with online compartmentalization.

• DOI: 10.1021/ac500365r
• 发表时间: 2014-04-01
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Volny, Michael;Rolfs, Joelle;Hakimi, Bejan;Frycak, Petr;Schneider, Thomas;Liu, Dingsheng;Yen, Gloria;Chiu, Daniel T.;Turecek, Frantisek
• 通讯作者: Turecek, Frantisek