CN110988102A - Visual sheath-flow-free single-cell mass spectrometry system - Google Patents
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1006—Investigating individual particles for cytology
Abstract
A visualized sheath-flow-free single-cell mass spectrometry system belongs to the field of mass spectrometry. The invention utilizes the equal-inner-diameter gold-plated capillary spray needle with narrow inner diameter and the pneumatic hydraulic sample introduction device to realize the mass spectrum analysis of single cells without sheath flow (without sheath liquid or sheath gas), an online visual microscopic system can observe the state of the single cells at the tip of the spray needle when participating in electrospray, further verify that each peak obtained by mass spectrum detection is from a single cell, and correspond the morphological information of the single cell to the signal characteristics of the mass spectrum thereof, thereby providing more reference information for the deep research of the heterogeneity of the single cell. The invention only adopts the gold-plated capillary spray needle with the equal inner diameter and the narrow inner diameter to carry out the single-cell spray mass spectrometry, does not need to add sheath flow to assist the spraying, does not need cell marking, improves the reliability of the single-cell mass spectrometry detection, and is a simple, reliable, high-throughput, high-sensitivity and high-coverage single-cell mass spectrometry analysis method.
Description
Technical Field
The invention relates to a visualized sheath-flow-free single-cell mass spectrometry system, and belongs to the field of mass spectrometry.
Background
Cells, as the basic building blocks of an organism, contain abundant biological information such as gene expression, interactions between cells, and metabolite responses caused by environmental changes. The traditional cell analysis method takes population cells as research objects to obtain average chemical information. With the development of analysis technology, a great deal of cell research shows that heterogeneity exists widely among cells due to the influence of genetic genes and external environment, and the difference among single cells has important value in the research of drug metabolism of cells, communication among cells and pathogenesis of diseases. However, the heterogeneity of cells is covered by the analysis of cell populations, and the analysis of cell differentiation cannot be realized, so that the single cell analysis is more and more focused by researchers, and becomes an important field in biological analysis.
At present, the flow cytometry commonly used is to analyze single cells based on fluorescence, and has higher flux. However, when performing multicolor fluorescence analysis, the number of detection channels is greatly limited (currently, 50 channels and 48 colors at most) due to the problem of overlapping fluorescence emission spectra, and the amount of single-cell information obtained is limited. The mass cytometry and mass spectrometer combined with the flow cytometry and the mass spectrometer is based on metal element marks, and the inductive coupling plasma mass spectrometry is used for flow cytometry detection, so that more than 30 signals can be obtained for each cell. However, both fluorescence-based flow cytometry and metal-element-based mass cytometry require labeling of the cell sample with fluorophores or metal elements. Both of the above methods have limited assay coverage and complicated sample pre-treatment resulting in intracellular sample loss, which are not conducive to single cell full-component analysis. Therefore, the development of a single cell detection technology without labeling, high throughput and high coverage is an urgent need for single cell analysis.
The organic mass spectrum has higher sensitivity, no need of marking, capability of simultaneously detecting a plurality of substances and good structure identification capability, and is very suitable for single cell detection and analysis. There are a number of mass spectrometry ion sources available today that are capable of ionizing different types of samples. The electrospray ion source is an atmospheric pressure ion source, can ionize single living cell components under the atmospheric pressure condition, and does not need complex sample pretreatment. Based on the nanoliter electrospray mass spectrometry technology for directly extracting or extracting the content of the single cell, signals of hundreds of metabolites can be detected by one-time sample injection, but the requirement on the operation technology is high, and the single cell detection flux is limited. The electrospray ionization mass spectrometry technology based on single-cell integral sample introduction is usually accompanied with the cracking in a single-cell tube and the auxiliary ionization process of sheath gas and sheath liquid, and the links inevitably cause the dilution and higher noise signals of a single-cell sample, and the signal-to-noise ratio of a single-cell mass spectrometry detection signal is seriously reduced. Therefore, on the basis of ensuring high throughput and high coverage of single cell detection, the establishment of a high-sensitivity single cell detection system and method has important significance.
The invention refers to the technical strategy of dispersing cell suspension by flow cytometry to ensure single cell sample introduction, and combines the characteristics of high sensitivity of nanoliter electrospray and high sample economy to establish a set of high-throughput, high-coverage and high-sensitivity visual sheath-flow-free single-cell mass spectrometry system. Under microscopic visualization, the invention only adopts one gold-plated capillary spray needle with the same inner diameter and the narrow inner diameter to carry out single-cell mass spectrometry detection and analysis, does not need to use sheath flow (sheath gas and sheath liquid) for auxiliary spraying, and is a simple, reliable, high-throughput, high-coverage and high-sensitivity single-cell mass spectrometry analysis method. The visual microscopic system can capture the participation state of cells in the spraying process of the tip of the spray needle by means of a high-speed camera shooting technology, and each peak obtained by mass spectrum detection can be in one-to-one correspondence with the imaged cells, so that the fact that each peak obtained by mass spectrum detection is from a single cell is more fully proved, and necessary system guarantee is provided for the relation between the cell morphology and the mass spectrum signal characteristics of the cell morphology.
Disclosure of Invention
The invention aims to overcome the defects of the existing single-cell mass spectrum detection technology and provide a visual sheath-flow-free single-cell mass spectrum analysis system. The method uses the gold-plated capillary spray needle with the equal inner diameter and the narrow inner diameter, improves the reliability of single cell sample introduction, is not easy to block, does not need sheath flow auxiliary spraying and sample marking, has simple device, can record the single cell state participating in electrospray in real time by a visual microscopic system, and corresponds the cell morphological information and the mass spectrum signal characteristics thereof one to one. Reliable single-cell high-throughput, high-coverage and high-sensitivity mass spectrum detection and analysis are realized.
In order to achieve the above object, the present invention adopts the following technical solutions.
A visualized sheath-flow-free single-cell mass spectrometry system is characterized by comprising a narrow-inner-diameter capillary spray needle (1) with equal inner diameter, a mass spectrometry detection system (2) and a visualized microscopic system; the inner diameters of the capillary spray needles (1) with the narrow inner diameters are equal, one end of each capillary spray needle is a tip, the tips are gradually thinned from the outer surface wall thickness and have the same inner diameter, and a gold layer with the submicron thickness is sprayed on the outer surface of the tip of each capillary spray needle with the narrow inner diameter;
the centrifugal tube (3) is arranged in the small steel cylinder (4), the cell suspension liquid is filled in the centrifugal tube (3), the high-pressure gas cylinder is connected with the centrifugal tube (3) through the high-pressure gas tube, and the port of the high-pressure gas tube is positioned above the cell suspension liquid in the centrifugal tube (3); one end of the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter is connected with a cell suspension liquid in the centrifuge tube (3), the other end of the capillary spray needle (1) with the equal inner diameter, namely the tip plated with the gold layer, is opposite to a mass spectrum inlet of the mass spectrum detection system (2), and a gap is reserved between the tip plated with the gold layer and the mass spectrum inlet to be used as a spray needle of an ion source in the mass spectrum detection system (2); the gold layer of the narrow-inner-diameter equal-inner-diameter capillary tip is connected with a high-voltage power supply, and high-voltage electricity is applied to the gold-plated tip to perform sample electrospray;
the side of the tip of the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter is provided with a visual microscopic system, the visual microscopic system comprises an objective lens (5), a lens barrel (6), a high-speed camera (7) and a light source (8), the objective lens (5) is sequentially connected with the lens barrel (6) and the high-speed camera (7) from the upper side of the tip of the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter, the objective lens (5), the lens barrel (6) and the high-speed camera (7) are integrally positioned on the tip of the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter, the light source (8) is positioned on the side of the tip of the spray needle, and the connecting line of the tip of the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter, the objective lens (5), the lens barrel; the visual microscopic system can capture the participation state of cells in the spraying process of the spray needle tip, each peak obtained by mass spectrum detection can be in one-to-one correspondence with the imaged cells, and the fact that each peak obtained by mass spectrum detection is from a single cell is fully proved.
To ensure cell viability and to allow direct access to mass spectrometry, cells were suspended in a mass-compatible isotonic solution and placed in a centrifuge tube (3).
The system is adopted to carry out visualized sheath-flow-free single-cell mass spectrometry, and is characterized by comprising the following steps:
(1) cutting a section of elastic quartz capillary tube with a narrow inner diameter, removing a polyimide coating at one end of the capillary tube, performing wet etching on the quartz capillary tube into tips with the same inner diameter, and plating a layer of gold on the tips with the same inner diameter by a film plating machine to prepare gold-plated capillary spray needles with the narrow inner diameter and the same inner diameter;
(2) preparing cell suspension, suspending cells in isotonic solution compatible with mass spectrum, and uniformly blowing to prepare cell suspension for later use;
(3) a sample introduction system is set up, the prepared cell suspension liquid is put into a centrifuge tube (3), and a high-pressure gas cylinder is connected with the centrifuge tube (3);
(4) one end of the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter is directly connected with the cell suspension liquid in the centrifuge tube (3), the gold-plated tip at the other end is opposite to the mass spectrum inlet of the mass spectrum detection system (2), high voltage is applied to the gold-plated tip, and the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter is used as a spray needle of an ion source in the mass spectrum detection system (2) for electrospray.
(5) A visual microscopic system is built, an objective lens (5) is connected with a lens cone (6), the lens cone (6) is connected with a high-speed camera (7), the whole body is located on one side of the tip of a gold-plated capillary needle (1) with the narrow inner diameter and the equal inner diameter, and a light source (8) is located on the other side of the tip of the needle.
(6) Opening a high-pressure gas cylinder, introducing nitrogen into a small steel cylinder (4), driving the cell suspension in the centrifugal tube (3) to enter a narrow-inner-diameter equal-inner-diameter capillary spray needle (1), performing electrospray on the tip of the narrow-inner-diameter equal-inner-diameter capillary spray needle (1), and entering mass spectrometry for detection and analysis; and simultaneously, a visual microscopic system is adopted to capture the participation state of cells in the spraying process of the tip of the spray needle.
Wherein, the inner diameter of the quartz capillary in the step (1) of the invention is 10-50 μm, and the thickness of the gold layer plated on the tip of the equal inner diameter by the film plating machine is submicron. The concentration of the isotonic solution (such as ammonium formate aqueous solution, ammonium acetate aqueous solution, etc.) in the step (2) is 10-250mmol/L, the diameter of the cells is 10-50 μm (only single cells are allowed to flow in turn in the inner diameter of a quartz capillary), and the concentration of the cell suspension is 1 x 104-1×106Per mL; the pressure of the high-pressure gas cylinder in the step (3) is 50-1000 psi. The high voltage in the step (4) is a voltage capable of electrospraying the cell suspension, and is generally not higher than 2kv, and preferably 1.2-1.4kv, and the detection mode of the mass spectrometry detection system can be a positive ion mode or a negative ion mode. And (6) capturing the participation state of the cells in the spraying process of the tip of the spray needle by using a high-speed camera shooting technology.
Under a high-speed microscopic system, the invention does not add sheath flow to assist spraying, only adopts a capillary spray needle with a narrow inner diameter and an equal inner diameter, and is a simple and reliable single-cell mass spectrometry analysis method. Due to the adoption of the technical scheme, the invention has the following advantages:
(1) the device is simple: the single-cell mass spectrometry is carried out only by adopting the equal-inner-diameter capillary spray needle with narrow inner diameter without adding sheath flow to assist spraying, and the method is a simple single-cell mass spectrometry method and is easy to master by operators.
(2) Online microscopic visualization: the participation state of cells in the spraying process of the tip of the spray needle can be captured, each peak obtained by mass spectrum detection can be in one-to-one correspondence with the imaged cells, so that the fact that each peak obtained by mass spectrum detection is from a single cell is more fully proved, and necessary system guarantee is provided for the relation between the cell morphology and the mass spectrum signal characteristics of the cell morphology.
(3) High coverage and high sensitivity: the method directly performs electrospray mass spectrometry on the whole single cell, can detect metabolites in the whole cell, does not dilute components in the cell, has the advantage of high coverage, and is favorable for realizing comprehensive single cell analysis. The method does not add sheath flow to assist spraying, reduces noise signals, improves the signal-to-noise ratio of mass spectrum detection signals, and has the advantage of high sensitivity.
(4) High flux: in the method, cells sequentially enter the gold-plated capillary spray needles with the equal inner diameters, flow type single cell analysis is realized, and the capillary spray needles with the equal inner diameters are used for carrying out electrospray, so that the method has the advantages of being difficult to block compared with a drawn conical capillary tip and having high-throughput single cell mass spectrometry.
(5) No marking is required: the cell sample does not need any label before detection and analysis, the problem that fluorescence and metal element labels are needed before flow cytometry and mass flow cytometer analysis is solved, and hundreds of metabolites of a single cell can be detected without labels at the same time.
(6) The cell pretreatment process is simple: the method only suspends cells in isotonic solution compatible with mass spectrum, can maintain the activity of the cells within a certain time, can promote electrospray ionization and improve the detection sensitivity.
Drawings
FIG. 1 is a schematic diagram of an apparatus for visualizing single-cell self-spray mass spectrometry detection according to the present invention (the size in the diagram does not represent the actual size ratio);
FIG. 2 is a total ion flow diagram for mass spectrometric detection of A549 cells in an embodiment of the invention;
FIG. 3 is a single A549 cell assay mass spectrum in accordance with an embodiment of the invention;
FIG. 4 shows the A549 cell status of an equi-inner diameter gold-plated capillary nozzle in electrospray photographed by a high-speed camera according to an embodiment of the present invention.
1 equal-inner-diameter gold-plated capillary spray needle with narrow inner diameter, 2 mass spectrum detection system, 3 centrifuge tube, 4 small steel cylinder, 5 objective lens, 6 lens cone, 7 high-speed camera and 8 light source.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The embodiment of the invention provides a visualized sheath-flow-free single-cell mass spectrometry system, and the invention is described in detail by taking an example of manufacturing an equal-inner-diameter gold-plated capillary spray needle by an elastic quartz capillary with an outer diameter of 360 microns and an inner diameter of 16 microns and carrying out high-throughput, high-coverage and high-sensitivity visualized sheath-flow-free single-cell detection and analysis on A549 cells, and the specific implementation steps are as follows:
the apparatus is shown in FIG. 1.
(1) Cutting a section of elastic quartz capillary tube with the length of about 30cm, removing a polyimide coating at one end of the capillary tube, etching the quartz capillary tube into tips with the same inner diameter by a wet method, plating a submicron-level thick gold layer on the tips with the same inner diameter by a low-temperature magnetron sputtering instrument to prepare a gold-plated capillary nozzle needle (1) with the same inner diameter and the narrow inner diameter, wherein the gold-plated length of the tips with the same inner diameter is 2 cm;
(2) digesting A549 cells from culture flask with trypsin, centrifuging, suspending in 125mmol/L ammonium formate aqueous solution, and gently blowing to obtain product with density of 2 x 105Cell suspension per mL for use;
(3) a sample injection system is set up, 100 mu L of the prepared A549 cell suspension is placed in a 200 mu L centrifuge tube (3) and is placed in a small steel cylinder (4), and a high-pressure gas cylinder is connected with the small steel cylinder (4);
(4) connecting the equal-inner-diameter gold-plated capillary spray needle (1) with a narrow inner diameter with a mass spectrum detection system (2), enabling the equal-inner-diameter gold-plated capillary spray needle (1) with the narrow inner diameter to work as an ion source spray needle of the mass spectrum detection system (2), adjusting the distance between the tip (1) of the equal-inner-diameter gold-plated capillary spray needle with the narrow inner diameter and a mass spectrum inlet to be about 2mm by using a three-dimensional adjusting platform, and applying high-voltage electricity to the gold-plated tip through a copper wire;
(5) opening a high-pressure gas cylinder, introducing nitrogen with the pressure of 800psi into a small steel cylinder (4), driving cell suspension in the small steel cylinder (4) to enter an equal-diameter gold-plated capillary spray needle (1) with a narrow inner diameter, applying high voltage of 1.3kV to the equal-diameter gold-plated capillary spray needle (1) with the narrow inner diameter for electrospray, detecting metabolites in A549 single cells in a mass spectrum negative ion mode, obtaining a total ion flow diagram (see figure 2) of high-flux and high-sensitivity mass spectrum detection of the A549 cells and a mass spectrogram (see figure 3) of the single A549 cells, wherein a pulse signal in the total ion flow diagram represents that one A549 cell is successfully detected.
(6) A visual microscopic system is built, an objective lens (5) is connected with a lens cone (6), the lens cone (6) is connected with a high-speed camera (7), the whole body is located right above the tip of a gold-plated capillary nozzle needle (1) with the narrow inner diameter and the equal inner diameter, a light source (8) is located on the side face of the tip of the nozzle needle, the participation state of cells in the spraying process of the tip of the nozzle needle is captured (see figure 4), each peak obtained through mass spectrum detection corresponds to an imaged cell one by one, and it is fully proved that each peak obtained through mass spectrum detection is from a single cell.
It should be noted that, because the method and the system are based on the same concept as the method embodiment of the present invention, the details of the cooperation, the execution process, and the like between the units in the method and the system may refer to the description in the method embodiment of the present invention, and are not described herein again.
The method and the system for detecting the visualized single-cell self-spraying mass spectrum provided by the embodiment of the invention are described in detail, the embodiment of the invention and the application of the obtained product are explained by applying specific examples, and the description of the embodiment is only used for helping to understand the method and the core idea of the invention; meanwhile, for a person skilled in the art, according to the idea of the present invention, the specific implementation manner and the application range may be changed, and the changes and the modifications belong to the protection scope of the present invention; in view of the above, the present disclosure should not be construed as limiting the invention.
Claims (9)
1. A visualized sheath-flow-free single-cell mass spectrometry system is characterized by comprising a narrow-inner-diameter capillary spray needle (1) with equal inner diameter, a mass spectrometry detection system (2) and a visualized microscopic system; the inner diameters of the capillary spray needles (1) with the narrow inner diameters are equal, one end of each capillary spray needle is a tip, the tips are gradually thinned from the outer surface wall thickness and have the same inner diameter, and a gold layer with the submicron thickness is sprayed on the outer surface of the tip of each capillary spray needle with the narrow inner diameter;
the centrifugal tube (3) is arranged in the small steel cylinder (4), the cell suspension liquid is filled in the centrifugal tube (3), the high-pressure gas cylinder is connected with the centrifugal tube (3) through the high-pressure gas tube, and the port of the high-pressure gas tube is positioned above the cell suspension liquid in the centrifugal tube (3); one end of the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter is connected with a cell suspension liquid in the centrifuge tube (3), the other end of the capillary spray needle (1) with the equal inner diameter, namely the tip plated with the gold layer, is opposite to a mass spectrum inlet of the mass spectrum detection system (2), and a gap is reserved between the tip plated with the gold layer and the mass spectrum inlet to be used as a spray needle of an ion source in the mass spectrum detection system (2); the gold layer of the narrow-inner-diameter equal-inner-diameter capillary tip is connected with a high-voltage power supply, and high-voltage electricity is applied to the gold-plated tip to perform sample electrospray;
the side of the tip of the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter is provided with a visual microscopic system, the visual microscopic system comprises an objective lens (5), a lens barrel (6), a high-speed camera (7) and a light source (8), the objective lens (5) is sequentially connected with the lens barrel (6) and the high-speed camera (7) from the upper side of the tip of the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter, the objective lens (5), the lens barrel (6) and the high-speed camera (7) are integrally positioned on the tip of the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter, the light source (8) is positioned on the side of the tip of the spray needle, and the connecting line of the tip of the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter, the objective lens (5), the lens barrel; the visual microscopic system can capture the participation state of cells in the spraying process of the tip of the spray needle.
2. A visual sheath-free single-cell mass spectrometry system according to claim 1, wherein to ensure cell viability and direct access to mass spectrometry, cells are suspended in a mass-compatible isotonic solution and placed in a centrifuge tube (3).
3. The system of claim 1, wherein the visualization microscopy system captures participation of the cells during the spraying process at the tip of the needle, and allows one-to-one correspondence of each peak detected by mass spectrometry with the imaged cells, sufficient to demonstrate that each peak detected by mass spectrometry is from a single cell.
4. A method for single cell mass spectrometry using the system of any one of claims 1 to 3, comprising the steps of:
(1) cutting a section of elastic quartz capillary with a narrow inner diameter, removing a polyimide coating at one end of the capillary, performing wet etching on the quartz capillary to form a tip with the same inner diameter, and plating a layer of gold on the tip with the same inner diameter by using a film plating machine to form a capillary spray needle with the same inner diameter and a narrow inner diameter;
(2) preparing cell suspension, suspending cells in isotonic solution compatible with mass spectrum, and uniformly blowing to prepare cell suspension for later use;
(3) a sample introduction system is set up, the prepared cell suspension liquid is put into a centrifuge tube (3), and a high-pressure gas cylinder is connected with the centrifuge tube (3);
(4) one end of a capillary spray needle (1) with the equal inner diameter and the narrow inner diameter is directly connected with a cell suspension liquid in a centrifuge tube (3), the gold-plated tip at the other end is opposite to a mass spectrum inlet of a mass spectrum detection system (2), high voltage is applied to the gold-plated tip, and the capillary spray needle (1) with the equal inner diameter and the narrow inner diameter is used as a spray needle of an ion source in the mass spectrum detection system (2) for electrospray;
(5) a visual microscopic system is built, an objective lens (5) is connected with a lens cone (6), the lens cone (6) is connected with a high-speed camera (7), the whole body is positioned on one side of the tip of a capillary spray needle (1) with the narrow inner diameter and the equal inner diameter, and a light source (8) is positioned on the other side of the tip of the spray needle;
(6) opening a high-pressure gas cylinder, introducing nitrogen into a small steel cylinder (4), driving the cell suspension in the centrifugal tube (3) to enter a narrow-inner-diameter equal-inner-diameter capillary spray needle (1), performing electrospray on the tip of the narrow-inner-diameter equal-inner-diameter capillary spray needle (1), and entering mass spectrometry for detection and analysis; and simultaneously, a visual microscopic system is adopted to capture the participation state of cells in the spraying process of the tip of the spray needle.
5. The method of claim 4, wherein in step (1) of the invention, the inner diameter of the quartz capillary is 10-50 μm, and the thickness of the gold layer coated by the coater at the tip with the same inner diameter is submicron.
6. The method according to claim 4, wherein the concentration of the isotonic solution of the step (2) is 10 to 250mmol/L, the diameter of the cells is 10 to 50 μm, only single cells are allowed to flow through the inner diameter of the quartz capillary in sequence, and the concentration of the cell suspension is 1X 104-1×106one/mL.
7. The method of claim 4, wherein the pressure of the high pressure gas cylinder in step (3) is 50-1000 psi.
8. The method according to claim 4, wherein in step (4), the high voltage is a voltage capable of electrospraying the cell suspension, generally not higher than 2kv, preferably 1.2-1.4kv, and the detection mode of the mass spectrometric detection system can be either positive or negative ion mode.
9. The method according to claim 4, wherein the participation of the cells in the spraying process of the tip of the needle is captured by using a high-speed camera technique in the step (6).
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| CN111397985A (en) * | 2020-04-14 | 2020-07-10 | 清华大学 | Single cell mass spectrometry |
| CN111982789A (en) * | 2020-08-21 | 2020-11-24 | 中国科学院生态环境研究中心 | High-throughput detection method of metal ions and metal nanoparticles based on single-cell enrichment and single-cell mass spectrometry |
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