Organic mass spectrum flow cytometry technology

Abstract

The invention discloses an organic mass spectrum flow cytometry technology which comprises the following steps: preparation of a protein labeled flow cytometry probe and a cell labeling method, construction of an organic mass spectrum flow cytometry device, mass spectrum flow cytometry detection of a cell suspension sample, and extraction and analysis of protein labels and metabolite information in a single cell mass spectrum. The technology is high in universality, the device is simple, and rapid, efficient, high-throughput and high-sensitivity sample analysis can be achieved. The flow mass spectrometry probe marks a large number of organic tags on target proteins, so that the detection sensitivity reaches the single cell level, qualitative and quantitative analysis of multiple target proteins and a large number of metabolites at the single cell level can be realized, and a large number of single cell level information is obtained. The technology has wide application prospects in the fields of cell identification, cell typing, research on characteristic difference substances in cells, tumor diagnosis, systematic biology research and the like.

Description

Technical field [0001] The present invention relates to a flow analysis technique based on the organic mass spectrometry in single cells, and specifically, a method of preparation of general protein-labeled fluid mass spectrometry and cell marking method, cellular single dispersion and arrangement technique, single cells Mass spectrometry detection of ionized interface design, as well as a single-cell level protein and metabolite detection and quantitative method based on mass spectrometry signals and its application in single cell identification and destination. Background technique [0002] Cells are the basic unit of life activities, and a major research content in life analysis is to analyze the mechanism of biological regulation on the cellular level. Single-cell analysis refers to monitoring life activities at a single cell or even less, its main analytical object is an important biomolecule in a single cell, such as DNA, RNA, protein and small molecular metabolic products,...

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Problems solved by technology

Literature (Y.Wang, R.Du, Qiao and B.Liu.Chem.Commun., 2018, 54, 9659-9662 and W.Ma, S.Xu, H.Nie, B.Hu, Y.Bai and H .Liu.Chem.Sci., 2019, 10, 2320-2325) reported typical organic mass spectrometry tags that can label macromolecules in cells, but the sensitivity of this type of probes has not yet reached the level of single cells, and Dissociation methods require special chemical reactions or lasers, so it is difficult to meet the needs of online rapid dissociation for single-cell flow analysis

Literature (G.Li, S.Yuan, S.Zheng, Y.Liu and G.Huang.Anal.Chem., 2018, 90, 3409-3415 and E.K.Neumann, T.J.Comi, S.S.Rubakhin and J.V.Sweedler.Angew. Chem.Int.Ed.2019, 58, 5910–5914) reported a typical ionization method and the design of the ionization device that can realize the release of single-cell content. However, the method is complicated to operate and the analysis time of a single cell is very long. Difficult to meet the speed requirements of streaming analysis

Literature (Q. Huang, S. Mao, M. Khan, L. Zhou and J. Lin. Chem. Commun., 2018, 54, 2595-2598 and H. Yao, H. Zhao, X. Zhao, X. Pan, J. Feng, F. Xu, S. Zhang and X. Zhang. Anal. Chem., 2019, 91, 9777-9783) reported an ionization interface that can initially realize flow analysis, which can detect metabolite information but Simultaneous detection of proteins cannot be achieved

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