Single cell mass spectrometry method

A mass spectrometry and single-cell technology, applied to the analysis of materials, individual particle analysis, particle and sedimentation analysis, etc., can solve the problems of reducing the convenience of operation, not conducive to popularization and application, and increasing sampling costs, so as to achieve convenient implementation and low cost. Low, easy-to-operate effect

Active Publication Date: 2020-07-10
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

As a result, the cost of sampling is greatly increased, the convenience of operation is reduced, and it is not conducive t

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Embodiment 1

[0054] Lipid substance analysis in single MCF7 cell of embodiment 1

[0055] Cell fixation: the cultured MCF7 cells (7-8 μm) were transferred to PBS, centrifuged at 1200 rpm, and the cells were resuspended in phosphate buffered saline (PBS) containing 2.5% glutaraldehyde. After 20 minutes, Cells were collected by centrifugation and resuspended in water to obtain a cell suspension.

[0056] Preparation of capillary glass tube: use a P1000 needle puller to draw a capillary glass tube with a tip diameter of 5 μm.

[0057] Mass Spectrometry:

[0058] (1) Add 0.5 μL of cell suspension into the capillary glass tube from the end of the capillary glass tube, select the capillary glass tube containing 1 cell under the microscope, and use -1.2kV voltage to migrate a single MCF7 cell to the front of the capillary glass tube.

[0059] (2) Wait for the liquid in the capillary glass tube to evaporate completely.

[0060] (3) A voltage of -+1.8 kV is applied while dropping auxiliary liqui...

Embodiment 2

[0062] Lipid analysis of MCF7 cells after embodiment 2 single photochemical derivation

[0063] Cell fixation: transfer the cultured cells to PBS. Using 1200 rpm centrifugation, the cells were resuspended in phosphate buffered saline (PBS) containing 2.5% glutaraldehyde, and after 20 minutes, centrifuged, the cells were collected and resuspended in water.

[0064] Photochemical derivation of cells: In the cell suspension, a solution of 2-acetylpyridine was added to a final concentration of 100 mM. In a quartz reaction box (such as figure 2 Shown) in, carry out ultraviolet photochemical reaction, used ultraviolet lamp is 254nm, and reaction time is 8 minutes.

[0065] Preparation of capillary glass tube: use a P1000 needle puller to draw a capillary glass tube with a tip diameter of 5 μm.

[0066] Mass Spectrometry:

[0067] (1) Add 0.5 μL of cell suspension (1-5 cells) into the capillary glass tube from the end of the capillary glass tube, select the capillary glass tube ...

Embodiment 3

[0072] Example 3 Structural identification of carbon-carbon double bond isomers of glycerophosphorylcholine (PC) in MCF7 cells after single photochemical derivatization

[0073] Cell fixation: transfer the cultured cells to PBS. Using 1200 rpm centrifugation, the cells were resuspended in phosphate buffered saline (PBS) containing 2.5% glutaraldehyde, and after 20 minutes, centrifuged, the cells were collected and resuspended in water.

[0074] Photochemical derivation of cells: In the cell suspension, a solution of 2-acetylpyridine was added to a final concentration of 100 mM. In a quartz reaction box (such as figure 2 Shown) in, carry out ultraviolet photochemical reaction, used ultraviolet lamp is 254nm, and reaction time is 8 minutes.

[0075] Preparation of capillary glass tube: use a P1000 needle puller to draw a capillary glass tube with a tip diameter of 5 μm.

[0076] Mass Spectrometry:

[0077] (1) Add 0.5 μL of cell suspension (1-5 cells) into the capillary gla...

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Abstract

The invention provides a single cell mass spectrometry method. The method comprises the following steps: carrying out cell immobilization and dilution treatment on cells to be detected so as to obtaina pretreated cell suspension liquid; injecting the pretreated cell suspension liquid into an inner cavity from a tail end of a capillary tube, wherein an electrode is inserted into the inner cavity;applying a voltage to the electrode so that the cells migrate to a capillary tip; completely evaporating the liquid in the capillary tube; dropwise adding an auxiliary liquid to the capillary tip, enabling the auxiliary liquid to enter the capillary tube to be in contact with the cells, extracting a target object; and meanwhile, applying the voltage to the electrode to perform electrospray so thatthe auxiliary liquid containing the target object is ionized, sprayed out from the tip and entering a sample inlet of a mass spectrometer to perform mass spectrometry. According to the method, sampling and mass spectrometry of the single cell can be achieved without depending on a high-precision control platform, identification of a double bond position of unsaturated lipids in the single cell can be achieved in combination with a photochemical derivation process, and an application prospect is good.

Description

technical field [0001] The present invention relates to the field of biology. In particular, the present invention relates to single cell mass spectrometry methods. Background technique [0002] There will be large differences between individuals in the cell population of the same genotype, that is, cell heterogeneity. There are significant differences between cells in gene transcription, protein expression, metabolism and other aspects. Therefore, single-cell analysis is of great significance for the study of intercellular heterogeneity. A variety of technologies have been widely used in single-cell analysis, such as single-cell sequencing, fluorescence detection technology, microfluidic technology, flow cytometry, mass spectrometer, etc. Among them, mass spectrometers are widely used in single-cell analysis due to their high sensitivity, accurate structure identification, and quantitative analysis. However, the small size and volume of cells themselves pose great chall...

Claims

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Application Information

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IPC IPC(8): G01N1/28H01J49/04G01N27/62G01N15/10
CPCG01N1/28H01J49/0445G01N27/62G01N15/10
Inventor 欧阳证李自帅马潇潇
Owner TSINGHUA UNIV
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