Mass spectrometry sampling microfluidic device and mass spectrometry detection apparatus

Abstract

The application discloses a kind of mass spectrometry sampling microfluidic device for subcellular microregion of living single cell, including sample handling system and microfluidic system, microfluidic system is placed in the upper of sample handling system, wherein, microfluidic system includes positioning mechanism, flow injection pump and microfluidic probe fixed on positioning mechanism, positioning mechanism is used to control microfluidic probe motion in three-dimensional direction, the bottom of microfluidic probe has patterned structure, the preset position of patterned structure is connected to the mass spectrometry inlet pipe line outside microfluidic probe by channel;Sample handling system includes temperature-controllable support assembly, petri dish and detection mechanism, petri dish is placed on support assembly, petri dish is used to place cell sample and keep cell activity and handle sample position, the detection mechanism for real-time observation sample is provided below support assembly.The application can be used for the multifunctional sampling of subcellular domain on living single cell, realizes subcellular mass spectrometry online analysis.

Description

Technical Field

[0001] This application relates to the field of single-cell mass spectrometry sampling technology, and in particular to a microfluidic device and mass spectrometry detection equipment for mass spectrometry sampling of subcellular microregions of live single cells. Background Technology

[0002] Subcellular analysis of living single cells can provide information on the spatial distribution of substances, helping to detect the early occurrence and development of physiological and pathological processes at a subtle level, which is of great significance to fields such as biomedicine. Current subcellular analysis mainly uses optical methods, such as fluorescence and chemiluminescence, to determine the distribution of intracellular target substances. However, the number of species that can be detected simultaneously by these optical methods is limited, and the information of the substances to be detected needs to be known in advance in order to design the corresponding optical detection system.

[0003] Furthermore, mass spectrometry can simultaneously detect hundreds or even thousands of substances, possessing the ability to detect unknown substances. However, mass spectrometry requires injecting samples into the instrument for analysis, and spatial information of substances is lost during sampling and injection. Existing high-precision subcellular mass spectrometry methods include cell puncture sampling and mass spectrometry imaging methods (MALDI, SIMS, etc.). The former can only extract intracellular fluid substances such as cytoplasm and cannot sample fixed components such as cell membranes and the cytoskeleton. The latter requires sample pretreatment operations such as washing, drying, and cell fixation, which can lead to cell death before sampling and loss of some substances, thus requiring improvement in analytical accuracy and comprehensiveness. Therefore, it is necessary to develop high-precision mass spectrometry sampling methods to comprehensively sample target subcellular microregions from live single cells, thereby supporting subcellular mass spectrometry analysis. Summary of the Invention

[0004] Therefore, it is necessary to provide a microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells. The microfluidic device of the present invention for mass spectrometry sampling of subcellular microregions of live single cells enables sampling of subcellular domains on live single cells, providing support for online subcellular mass spectrometry analysis and laying the foundation for precise single-cell analysis.

[0005] One embodiment of this application provides a microfluidic device for mass spectrometry sampling of subcellular microregions of living single cells.

[0006] A microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells includes a sample manipulation system and a microfluidic system, wherein the microfluidic system is positioned above the sample manipulation system.

[0007] The microfluidic system includes a positioning mechanism, a flow injection pump, and a microfluidic probe fixed on the positioning mechanism. The positioning mechanism is used to control the movement of the microfluidic probe in three dimensions. The bottom of the microfluidic probe has a patterned structure for holding a target single cell. The patterned structure can move to contact the surface of the culture dish. The preset position of the patterned structure is connected to the mass spectrometer inlet tube outside the microfluidic probe through a channel.

[0008] The sample manipulation system includes a temperature-controlled support component, a culture dish, and a detection mechanism. The culture dish is placed on the support component, which is capable of moving along the X-axis and Y-axis. The culture dish is used to place cell samples, maintain cell viability, and manipulate the sample position. The detection mechanism for real-time observation of the sample is located below the support component.

[0009] In some embodiments, the patterned structure includes a first channel, a second channel, and a microchamber, the microchamber being used to hold a target single cell, the first channel and the second channel being respectively connected to the microchamber, and the first channel and the second channel being respectively used for injecting and effluent of solution.

[0010] In some embodiments, there are multiple first channels, and the multiple first channels are distributed in a divergent manner;

[0011] And / or, the number of the second channels is multiple, and the multiple second channels are distributed in a divergent manner.

[0012] In some embodiments, the patterned structure can realize multiple first channel injection solutions and multiple second channel effluent solutions, or the patterned structure can realize one first channel injection solution and multiple second channel effluent solutions, or the patterned structure can realize multiple first channel injection solutions and one second channel effluent solution.

[0013] In some embodiments, the first channel and the second channel are respectively located on opposite sides of the microchamber.

[0014] In some embodiments, the cell sample in the culture dish is planted at a density of 10-1. 2 ~10 4 pcs / square centimeter

[0015] In some embodiments, the positioning mechanism is an XYZ positioner, which is capable of movement in the X-axis, Y-axis and Z-axis directions.

[0016] In some embodiments, the sample handling platform further includes a detection mechanism, which is disposed below the support assembly. The detection mechanism is a microscope, and the objective lens of the microscope faces the support assembly.

[0017] In some embodiments, the support assembly is able to control the temperature of the culture dish to 37°C.

[0018] One embodiment of this application also provides a mass spectrometry detection device.

[0019] A mass spectrometry detection device includes a microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells.

[0020] This invention provides a microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells, which has the following advantages compared to traditional technologies:

[0021] 1. This invention samples the target subcellular domain of a living single cell by manipulating the fluid flow in the microcavity at the bottom of a microfluidic probe. It can simultaneously analyze a large number of components at specific subcellular locations of a single cell and has the ability to detect unknown substances.

[0022] 2. The fluid injected in this invention can be either a solvent capable of dissolving cellular components for extracting components from the target subcellular domain, or a non-damaging fluid for online collection of exocrine substances from the target subcellular domain. Except for some special applications (such as using a solvent to dissolve the cell nucleus for sampling), in most application scenarios, the cells sampled using this method can still survive and even regenerate, making it possible to sample and analyze multiple times from the same living cell, which is of great significance for life science research.

[0023] 3. This invention can be used to study the compositional differences in different parts of a single cell, thereby providing important information for research on early physiological and pathological processes in subtle ways.

[0024] 4. This invention can damage subcellular domains by injecting damaging substances and simultaneously perform mass spectrometry analysis. It can be used to explore changes in the composition near the wound before and after injury, and to study whether the composition of cells after repairing wounds and regenerating new limbs is different from before, thus revealing the properties of cells in depth. Attached Figure Description

[0025] To more clearly illustrate the technical solutions in the embodiments of this application, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0026] To gain a more complete understanding of this application and its beneficial effects, the following description will be provided in conjunction with the accompanying drawings. In the following description, the same reference numerals denote the same parts.

[0027] Figure 1 This is a schematic diagram of the overall structure of a microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells according to an embodiment of the present invention.

[0028] Figure 2 This is a schematic diagram of the microfluidic probe structure of a microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells according to an embodiment of the present invention.

[0029] Figure 3 This is a schematic diagram of the patterned structure of the microfluidic probe of the microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells according to an embodiment of the present invention.

[0030] Figure 4 This is a schematic diagram of the sampling fluid manipulation of the microfluidic probe in a microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells, according to an embodiment of the present invention.

[0031] Explanation of reference numerals in the attached figures

[0032] 10. Microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells; 100. Microfluidic system; 101. Microfluidic probe; 1011. Patterned structure; 1012. First channel; 1013. Second channel; 1014. Microchamber; 200. Sample manipulation system; 201. Support component; 202. Culture dish; 203. Detection mechanism; 20. Sample cell. Detailed Implementation

[0033] To make the above-mentioned objects, features, and advantages of the present invention more apparent and understandable, specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings. Many specific details are set forth in the following description to provide a thorough understanding of the present invention. However, the present invention can be practiced in many other ways different from those described herein, and those skilled in the art can make similar modifications without departing from the spirit of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0034] In the description of this invention, it should be understood that the terms "center," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," and "circumferential" indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this invention.

[0035] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0036] In this invention, unless otherwise explicitly specified and limited, the terms "installation," "connection," "linking," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components, unless otherwise explicitly limited. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0037] In this invention, unless otherwise explicitly specified and limited, "above" or "below" the second feature can mean that the first feature is in direct contact with the second feature, or that the first feature is in indirect contact with the second feature through an intermediate medium. Furthermore, "above," "over," and "on top" of the second feature can mean that the first feature is directly above or diagonally above the second feature, or simply that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature can mean that the first feature is directly below or diagonally below the second feature, or simply that the first feature is at a lower horizontal level than the second feature.

[0038] It should be noted that when an element is referred to as being "fixed to" or "set on" another element, it can be directly on the other element or there may be an intervening element. When an element is considered to be "connected to" another element, it can be directly connected to the other element or there may be an intervening element. The terms "vertical," "horizontal," "upper," "lower," "left," "right," and similar expressions used herein are for illustrative purposes only and do not represent the only possible implementation.

[0039] In the description of this invention, "several" means one or more, "multiple" means two or more, "greater than," "less than," and "exceeding" are understood to exclude the stated number, while "above," "below," and "within" are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly indicating the number of indicated technical features, or implicitly indicating the order of the indicated technical features.

[0040] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0041] This application provides a microfluidic device 10 for mass spectrometry sampling of subcellular microregions of living single cells, addressing at least one of the following problems: existing optical methods can only detect a limited number of species simultaneously and require prior knowledge of the substances to be detected in order to design corresponding optical probes; and mass spectrometry sampling and injection processes result in the loss of spatial information of substances or incomplete or inaccurate sampling. The microfluidic device 10 for mass spectrometry sampling of subcellular microregions of living single cells will be described below with reference to the accompanying drawings.

[0042] The microfluidic device 10 for mass spectrometry sampling of subcellular microregions of live single cells provided in this application embodiment is exemplary; please refer to [link to example]. Figure 1 As shown, Figure 1 This is a schematic diagram of the structure of a microfluidic device 10 for mass spectrometry sampling of subcellular microregions of live single cells, provided in an embodiment of this application. The microfluidic device 10 for mass spectrometry sampling of subcellular microregions of live single cells can be used to sample subcellular domains on live single cells, providing support for online subcellular mass spectrometry analysis and laying the foundation for precise single-cell analysis.

[0043] To more clearly illustrate the structure of the microfluidic device 10 for mass spectrometry sampling of subcellular microregions of live single cells, the following description, in conjunction with the accompanying drawings, will be provided on the microfluidic device 10 for mass spectrometry sampling of subcellular microregions of live single cells.

[0044] For example, please refer to Figure 1 As shown, Figure 1 This is a schematic diagram of the structure of a microfluidic device 10 for mass spectrometry sampling of subcellular microregions of live single cells, provided in an embodiment of this application.

[0045] An exemplary microfluidic device 10 for mass spectrometry sampling of subcellular microregions of live single cells includes a sample manipulation system 200 and a microfluidic system 100. The microfluidic system 100 is positioned above the sample manipulation system 200.

[0046] The microfluidic system 100 includes a positioning mechanism, a flow injection pump, and a microfluidic probe 101 fixed to the positioning mechanism. The positioning mechanism controls the movement of the microfluidic probe 101 in three dimensions, specifically, it can move along the X, Y, and Z axes. The bottom of the microfluidic probe 101 has a patterned structure 1011 for holding a target single cell. The patterned structure 1011 can move to contact the surface of the culture dish 202. The preset position of the patterned structure 1011 is connected to a mass spectrometry inlet pipe outside the microfluidic probe 101 via a channel. This application uses a flow injection pump to adjust the flow rate of each fluid stream and controls the type of solution injected into each channel by adjusting the pipe connection method, thereby controlling the relative position and size of the area through which the cell extract flows, and completing the sampling of the target subcellular domain of a single cell.

[0047] The sample manipulation system 200 includes a temperature-controlled support assembly 201, a culture dish 202, and a detection mechanism 203. The culture dish 202 is placed on the support assembly 201. The support assembly 201 is movable along the X-axis and Y-axis. The culture dish 202 is used to place cell samples, maintain cell viability, and manipulate the sample position. The detection mechanism 203 for real-time observation of the sample is located below the support assembly 201.

[0048] The aforementioned microfluidic device 10 for mass spectrometry sampling of subcellular microregions of live single cells, in use, places the cell sample in a culture dish 202 on a temperature-controlled support component 201 to ensure cell viability and manipulate the sample position; the microfluidic probe 101 is fixed to a positioning mechanism such as an XYZ positioner, allowing precise three-dimensional control of the microfluidic probe 101's position; the bottom of the microfluidic probe 101 has micropatterned grooves, within which microchambers 1014 can hold the target single cell in the culture dish 202, and the first channel 1012 and second channel 1013 at the edge are used for solution introduction. The fluid is driven by a flow injection pump, allowing for precise flow rate control. Different streams of solution are injected and flow through the target single cell in a laminar flow state within the microchamber 1014, dividing the cell into multiple subcellular microregions. Each stream of fluid flows through its corresponding subcellular domain and then into its corresponding outlet channel. One or more streams of fluid may be solutions that extract cell components (e.g., cell lysis buffer), which dissolve and sample the subcellular microregions through which they flow. The second channel 1013 corresponding to the sampling fluid is connected to the mass spectrometer inlet via tubing, thereby enabling online mass spectrometry analysis of the extracted subcellular domain samples.

[0049] In some embodiments, the support component 201 may be an XY stage.

[0050] In some of these embodiments, please refer to Figure 2 As shown, the patterned structure 1011 includes a first channel 1012, a second channel 1013, and a microchamber 1014. The microchamber 1014 is used to hold the target single cell, and the first channel 1012 and the second channel 1013 are respectively connected to the microchamber 1014. The first channel 1012 and the second channel 1013 are used for injecting and effluent of solution, respectively. For example, see... Figure 3 As shown, each first channel 1012 is used for injecting solution, and each second channel 1013 is used for effluent flowing out of solution. The sampling size and location can be adjusted by the solution injection method and flow rate; please refer to [reference needed]. Figure 4 As shown.

[0051] In some embodiments, there are multiple first channels 1012, and the multiple first channels 1012 are distributed in a divergent manner.

[0052] In some embodiments, there are multiple second channels 1013, and the multiple second channels 1013 are distributed in a divergent manner.

[0053] In some embodiments, the patterned structure 1011 can realize multiple first channels 1012 for injecting solution and multiple second channels 1013 for effluent, or the patterned structure 1011 can realize one first channel 1012 for injecting solution and multiple second channels 1013 for effluent, or the patterned structure 1011 can realize multiple first channels 1012 for injecting solution and one second channel 1013 for effluent.

[0054] In some embodiments, the first channel 1012 and the second channel 1013 are respectively disposed on opposite sides of the microchamber 1014.

[0055] In some embodiments, the cell sample seeding density in culture dish 202 is 10-1. 2 ~10 4 pcs / square centimeter

[0056] In some embodiments, the positioning mechanism is an XYZ positioner. The positioning mechanism is capable of movement in the X-axis, Y-axis, and Z-axis directions.

[0057] In some embodiments, the sample handling platform further includes a detection mechanism 203. The detection mechanism 203 is disposed below the support assembly 201, wherein the detection mechanism 203 is a microscope. (See attached image) Figure 1 As shown, the objective lens of the microscope is oriented toward the support assembly 201.

[0058] In some embodiments, the support component 201 is able to control the temperature of the culture dish 202 to 37°C.

[0059] One embodiment of this application also provides a mass spectrometry detection device.

[0060] A mass spectrometry detection device includes a microfluidic device 10 for mass spectrometry sampling of subcellular microregions of live single cells. The patterned structure 1011 of the microfluidic device 10 for mass spectrometry sampling of subcellular microregions of live single cells is connected via channels to the mass spectrometry inlet tubing of the mass spectrometry detection device outside the microfluidic probe 101. The preset positions are one or more second channels 1013.

[0061] When the microfluidic device 10 of the present invention is used for mass spectrometry sampling of subcellular microregions of living single cells, the process is as follows:

[0062] Step (1): Place the cell sample in the culture dish 202, culture the cell sample to the state to be detected, and then place the culture dish 202 on the sample manipulation system 200 to adjust the target cells to the appropriate position.

[0063] Step (2): Connect the first channel 1012 of the microfluidic probe 101 to the corresponding syringe, which contains the corresponding solution. The syringe is mounted on a flow injection pump and a non-damaging solution (e.g., cell buffer) is pre-injected to purge gas from the flow path.

[0064] Step (3): Cover the target single cell in the culture dish 202 with the bottom microchamber 1014 of the microfluidic probe 101, and start each flow injection pump to cause the fluid to flow through the target single cell in the culture dish 202 in a laminar flow state. Please refer to [link to relevant documentation]. Figure 3 As shown.

[0065] Step (4): Collect sample solutions from different outlets of the second channel 1013, and send the sample solution of interest into a mass spectrometer for analysis. Please refer to [link to relevant documentation]. Figure 3 As shown. The sampling size and location can be adjusted by the solution injection method and flow rate; please refer to [link / reference]. Figure 4 As shown.

[0066] Step (5): After sampling, if a cell-damaging solution was used: stop the injection and slowly aspirate the cell-damaging solution back into the tubing, allowing it to be pushed back by the non-damaging solution injected through other inlet channels. Then lift and remove the microfluidic probe and return the cells to the culture environment. If no cell-damaging solution was used, simply lift and remove the microfluidic probe.

[0067] In summary, the microfluidic device 10 of the present invention for mass spectrometry sampling of subcellular microregions of live single cells has the following advantages compared with the technical solutions of the traditional technology:

[0068] 1. This invention samples the target subcellular domain of a living single cell by manipulating the fluid flow within the microcavity 1014 at the bottom of the microfluidic probe. It can simultaneously analyze a large number of components at specific subcellular locations of a single cell and has the ability to detect unknown substances.

[0069] 2. The fluid injected in this invention can be either a solvent capable of dissolving cellular components for extracting components from the target subcellular domain, or a non-damaging fluid for online collection of exocrine substances from the target subcellular domain. Except for some special applications (such as using a solvent to dissolve the cell nucleus for sampling), in most application scenarios, the cells sampled using this method can still survive and regenerate, making it possible to sample and analyze multiple times from the same living cell, which is of great significance for life science research.

[0070] 3. This invention can be used to study the compositional differences in different parts of a single cell, thereby providing important information for research on early physiological and pathological processes in subtle ways.

[0071] 4. This invention can damage subcellular domains by injecting damaging substances and simultaneously perform mass spectrometry analysis. It can be used to explore changes in the composition near the wound after injury, and to study whether the composition of cells after wound repair and regeneration is different from before, thus revealing the properties of cells in depth.

[0072] In the above embodiments, the descriptions of each embodiment have different focuses. For parts not described in detail in a certain embodiment, please refer to the relevant descriptions in other embodiments.

[0073] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0074] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. A microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells, characterized in that, It includes a sample manipulation system and a microfluidic system, with the microfluidic system positioned above the sample manipulation system. The microfluidic system includes a positioning mechanism, a flow injection pump, and a microfluidic probe fixed on the positioning mechanism. The positioning mechanism controls the movement of the microfluidic probe in three dimensions. The bottom of the microfluidic probe has a patterned structure for holding a target single cell. The patterned structure can move to contact the surface of the culture dish. The preset position of the patterned structure is connected to a mass spectrometry inlet tube outside the microfluidic probe through a channel. The patterned structure includes a first channel, a second channel, and a microchamber. The microchamber is used to hold the target single cell. The first channel and the second channel are respectively connected to the microchamber and are used for injecting and effluent solutions, respectively. The sample manipulation system includes a temperature-controlled support component, a culture dish, and a detection mechanism. The culture dish is placed on the support component, which is capable of moving along the X-axis and Y-axis. The culture dish is used to place cell samples, maintain cell viability, and manipulate the sample position. The detection mechanism for real-time observation of the sample is located below the support component.

2. The microfluidic device for mass spectrometry sampling of subcellular microregions of living single cells according to claim 1, characterized in that, There are multiple first channels, and these multiple first channels are distributed in a divergent manner.

3. The microfluidic device for mass spectrometry sampling of subcellular microregions of living single cells according to claim 2, characterized in that, There are multiple second channels, and these multiple second channels are distributed in a divergent manner.

4. The microfluidic device for mass spectrometry sampling of subcellular microregions of living single cells according to claim 3, characterized in that, The patterned structure can realize multiple first channel injection solutions and multiple second channel outflow solutions, or the patterned structure can realize one first channel injection solution and multiple second channel outflow solutions, or the patterned structure can realize multiple first channel injection solutions and one second channel outflow solution.

5. The microfluidic device for mass spectrometry sampling of subcellular microregions of living single cells according to claim 3, characterized in that, The first channel and the second channel are respectively located on opposite sides of the microchamber.

6. The microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells according to any one of claims 1-5, characterized in that, The cell sample in the culture dish was planted at a density of 10. 2 ~10 4 pcs / square centimeter 7. The microfluidic device for mass spectrometry sampling of subcellular microregions of living single cells according to any one of claims 1-5, characterized in that, The positioning mechanism is an XYZ positioner, which is capable of moving in the X-axis, Y-axis and Z-axis directions.

8. The microfluidic device for mass spectrometry sampling of subcellular microregions of living single cells according to any one of claims 1-5, characterized in that, The sample handling platform also includes a detection mechanism, which is located below the support assembly. The detection mechanism is a microscope, and the objective lens of the microscope faces the support assembly.

9. The microfluidic device for mass spectrometry sampling of subcellular microregions of living single cells according to any one of claims 1-5, characterized in that, The support assembly can control the temperature of the culture dish to 37°C.

10. A mass spectrometry detection device, characterized in that, The microfluidic device for mass spectrometry sampling of subcellular microregions of live single cells, as described in any one of claims 1-9.

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