Microporous piezoelectric single cell sampling device and application

Abstract

The invention provides a microporous piezoelectric single cell sampling device and application, and belongs to the technical field of single cell detection. The microporous piezoelectric single cell sample introduction device comprises a sample treatment part, a carrier gas transmission channel and an aerosol transmission part, the sample treatment part comprises a microporous piezoelectric cell screening assembly, a microporous piezoelectric cell introduction assembly and a linkage circuit controller, and the sample treatment part is coaxially arranged at one end of the aerosol transmission part; the carrier gas transmission channel is communicated with the aerosol transmission part; the aerosol transmission part is conical; cells with different sizes can be screened on line according to cell sizes, and efficient introduction of specific target cells is realized; the device can generate low-speed aerosol micelles with an extremely low divergence angle under the condition of not depending on carrier gas, and the transmission efficiency of a cell sample is easy to improve through airflow regulation and control; according to the device, the generation of waste liquid is avoided, and the pollution to the environment is effectively reduced. The device can also be used in combination with an inductively coupled plasma mass spectrometer.

Description

Technical Field

[0001] The present invention relates to the technical field of single cell detection, and in particular to a micropore piezoelectric single cell sampling system and a single cell detection method. Background Art

[0002] Single-cell analysis is an important research field in recent years. It can reveal the individual characteristics of cells, help study the complexity of cells themselves and the differences between cells, and has very important biological value. Inductively coupled plasma mass spectrometry (ICP-MS) plays a key role in detecting the content and distribution of metal elements in single cells due to its excellent trace metal element analysis capabilities. In recent years, metal nanoparticles, especially functionalized metal nanoparticles, have been widely used in bioimaging, intracellular sensing, disease treatment and other directions due to their unique chemical and physical properties. However, there are few reports on their accumulation, distribution and heterogeneity in cells. In biomedical applications and toxicological analysis, the study of cellular uptake of nanoparticles plays a vital role. Therefore, it is of great significance to establish a high-throughput and high-precision single-cell analysis method to study the effects of nanoparticles on cells or organisms. However, the injection efficiency of conventional sample introduction systems based on pneumatic atomization is usually 1-10%, which means that most of the samples are lost with the removal of waste liquid. Inefficient injection systems are not conducive to the analysis of precious trace cell samples. In addition, the waste liquid generated also has the risk of polluting the environment. Therefore, an efficient sample injection system is very important for ICP-MS analysis instruments.

[0003] At present, some efficient sampling systems have been reported, such as the invention patent authorization announcement number US10497550 (B1) and CN207816913U disclose membrane desolvation device (Membrane Desolvation, MD); patent CN104931420B discloses an ultrasonic nebulization sampling system (Ultrasonic Nebulization, USN); patent CN217954234U and CN207233698U disclose electrothermal vaporization sampling system (Electrothermal Vaporization, ETV). Efficient sampling system is a constraint on the development of large-scale inorganic analytical instruments. The above-mentioned efficient sampling systems that have been reported can improve the sampling efficiency and enhance the analysis sensitivity, but they all have some disadvantages, such as MD is expensive (tens of thousands of dollars), residues are difficult to clean, and the device maintenance process is cumbersome; USN also has the problem of high price and residue cleaning, and the existing commercial USN has a high sampling rate and cannot meet the needs of trace sample analysis. ETV uses heat energy to convert analytes into gaseous species, and has a certain selectivity in sample introduction, but the analysis throughput is low, and it cannot introduce nanoparticles and cell type samples, and has not yet become a conventional sample introduction device for ICP-MS. Therefore, it is very necessary to develop a new type of high-throughput, high-injection efficiency and low-residue trace sample efficient introduction system.

[0004] In recent years, microporous piezoelectric atomization has been proven to be able to convert trace amounts of solution into uniform and smaller-sized aerosol micelles for spraying, and is a very effective atomization sampling method. Compared with conventional pneumatic atomization sampling systems, this method has lower costs and can produce low-speed aerosol micelles that are easy to control, providing a new idea for the development of new ICP-MS efficient sample introduction systems. However, the existing microporous piezoelectric atomization device cannot be directly used as a cell sampling system for ICP-MS analysis instruments. The reasons are: (1) Cells vary in size, and the microporous structure of existing microporous piezoelectric atomization sheets is usually fixed and the outlet aperture is much smaller than that of ordinary cells, resulting in the inability of cells to effectively pass through the micropores into the analysis instrument with the aerosol, and larger particles are easily accumulated in the pores to cause blockage; (2) The micropore edges of existing commercial microporous piezoelectric atomization sheets have irregular sharp edges, which can easily cause damage to cells in the process of passing through the micropores. Therefore, the present invention proposes to develop a new type of efficient cell biological sample introduction system based on a microporous piezoelectric cell atomization device for ICP-MS analysis. Summary of the invention

[0005] The purpose of the present invention is to address the above-mentioned deficiencies in the prior art and provide a microporous piezoelectric single-cell sampling device and application, which can screen cells of different sizes online according to cell size to achieve efficient introduction of specific target cells. The device has extremely high transmission efficiency and greatly reduces the amount of sample required for analysis of precious trace biological samples such as cells.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] The first object of the present invention is to provide a microporous piezoelectric single cell sampling device, which is used in the sample introduction system of an ICP-MS analyzer to introduce an atomized sample into an ICP matrix tube. The microporous piezoelectric single cell sampling device comprises a sample processing part, a carrier gas transmission channel and an aerosol transmission part. The sample processing part is coaxially arranged at one end of the aerosol transmission part, and the carrier gas transmission channel is connected to the aerosol transmission part; the aerosol transmission part is conical;

[0008] The sample processing part includes a microporous piezoelectric cell screening component, a microporous piezoelectric cell introduction component and a linkage circuit controller. The microporous piezoelectric cell screening component and the microporous piezoelectric cell introduction component are arranged at intervals and are sealed at the inlet end of the aerosol transmission part through a sealing fixing ring. The linkage circuit controller controls the piezoelectric effect of the microporous piezoelectric cell screening component and the microporous piezoelectric cell introduction component; the carrier gas transmission channel is arranged along the inner wall of the aerosol transmission part to generate a vortex carrier gas flow in the fog chamber cavity; the outlet end of the aerosol transmission part is connected to the ICP matrix tube.

[0009] Furthermore, the microporous piezoelectric cell screening component and the microporous piezoelectric cell introduction component are both formed by bonding a microporous piezoelectric metal sheet and a piezoelectric ceramic ring, and a microporous area is arranged at the center of the microporous piezoelectric metal sheet.

[0010] Furthermore, the distance between the microporous piezoelectric cell screening component and the microporous piezoelectric cell introducing component is 5 to 30 mm.

[0011] Furthermore, the diameter of the microporous piezoelectric metal sheet ranges from 3 to 50 mm.

[0012] Furthermore, the inlet diameter of the micropores on the microporous piezoelectric metal sheet ranges from 50 to 500 μm.

[0013] Furthermore, the diameter of the spray outlet of the micropores on the microporous piezoelectric metal sheet ranges from 10 to 100 μm.

[0014] Furthermore, the carrier gas transmission channel includes a carrier gas pipeline and a compensation gas pipeline, and the carrier gas pipeline and the compensation gas pipeline are radially symmetrically arranged on the inner wall of the aerosol transmission part along the microporous piezoelectric metal sheet.

[0015] Furthermore, the cavity volume of the aerosol transmission part is between 10 and 200 mL.

[0016] Furthermore, the microporous piezoelectric cell screening component and the microporous piezoelectric cell introduction component are integrally formed by 3D printing.

[0017] The second object of the present invention is to provide an inductively coupled plasma mass spectrometer comprising the above-mentioned microporous piezoelectric single-cell sampling device.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] (1) Compared with conventional pneumatic atomization injection systems and other high-efficiency injection systems, the microporous piezoelectric single-cell injection device provided by the present invention has the following characteristics: the core component of the device provided by the present invention, the microporous piezoelectric atomization sheet, is made by 3D printing in one piece, and the rounded microporous structure made in this way does not damage cells; cells of different sizes can be screened online according to cell size to achieve efficient introduction of specific target cells; and the device can generate low-speed aerosol micelles with extremely low divergence angles without relying on carrier gas, and it is easy to improve the transmission efficiency of cell samples by airflow regulation; the device avoids the generation of waste liquid and effectively reduces pollution to the environment.

[0020] (2) The present invention provides a micro-pore piezoelectric single-cell sampling device. The micro-amount and high-efficiency cell biological sample introduction system based on piezoelectric effect droplet injection can screen cell types based on particle size differences. The device has extremely high transmission efficiency, greatly reducing the amount of sample required for the analysis of precious trace biological samples such as cells. No waste liquid is generated during the sampling process, which is green and environmentally friendly.

[0021] (3) The microporous piezoelectric single-cell sampling device provided by the present invention can be used in conjunction with an inductively coupled plasma mass spectrometer to provide a novel analytical solution for element and isotope detection of trace cell samples. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 A schematic diagram of the structure of the microporous piezoelectric single-cell sampling device provided by the present invention;

[0023] Figure 2a A schematic diagram of the microporous structure of the microporous piezoelectric atomizer provided by the present invention in a longitudinal section;

[0024] Figure 2b A schematic diagram of the micropore area for cell entry provided by the present invention;

[0025] Figure 3 It is a schematic structural diagram of an inductively coupled plasma mass spectrometer including a microporous piezoelectric single cell sampling device provided by the present invention;

[0026] Figure 4a The low flow rate (10 μL min) of Example 1 of the present invention -1 ) Comparison of the number of cell events obtained by introducing the same number of blue cells into the inductively coupled plasma quadrupole mass spectrometer using micropore piezoelectric atomization; ICP-MS uses a high time resolution data acquisition mode with a dwell time of 10 milliseconds to continuously monitor 44 The horizontal axis represents the monitoring time, and the vertical axis represents the number of Ca ions detected in each 10 millisecond time window. 44 The signal intensity of Ca ion counting is in the form of pulse peaks, and the number of pulse peaks corresponds to the number of blue cells detected in that time period;

[0027] Figure 4b The number of cell events is obtained by introducing the same number of blue cells into the inductively coupled plasma quadrupole mass spectrometer using conventional pneumatic nebulization at a high flow rate; the ICP-MS uses a high time resolution data acquisition mode with a dwell time of 10 milliseconds to continuously monitor 44 The horizontal axis represents the monitoring time, and the vertical axis represents the number of ions detected in each 10 millisecond time window. 44 The signal intensity of Ca ion counting is in the form of pulse peaks, and the number of pulse peaks corresponds to the number of blue cells detected in that time period;

[0028] Figure 5 The number of cell events obtained by introducing the microporous piezoelectric atomization sample into the inductively coupled plasma quadrupole mass spectrometer after cell screening in Example 2 of the present invention; the ICP-MS adopts a high time resolution data acquisition mode with a dwell time of 10 milliseconds to continuously monitor 57 Fe ion signal, the horizontal axis represents the monitoring time, and the vertical axis represents the detected ion signal in each 10 millisecond time window. 57 The signal intensity of Fe ion counting is in the form of pulse peaks, and the number of pulse peaks corresponds to the number of red blood cells detected in that time period;

[0029] Figure 6 It is a comparison chart of the number and concentration of cells of two types of cells observed under microscopic observation before and after screening in Example 2. The counting method uses a traditional Neubauer cell counting plate to count the cells of the sample solution diluted 50 times with physiological saline before and after screening under an upright microscope. The number and concentration of cells can be obtained by multiplying the number of observed cells by the dilution factor. The horizontal axis represents the cell type and the vertical axis represents the number and concentration of cells. After screening, the number and concentration of red blood cells in the sample solution remain unchanged, while the number of epithelial cells decreases by more than two orders of magnitude, indicating that the microporous piezoelectric cell screening component effectively retains large-size epithelial cells and non-destructively and efficiently selectively introduces small-size red blood cells.

[0030] Figure 7 It is a circuit diagram controlled by the linkage circuit controller adopted in the embodiment.

[0031] In the figure: 1. Sample processing unit; 11. Microporous piezoelectric cell screening component; 12. Microporous piezoelectric cell introduction component; 13. Linkage circuit controller; 14. Microporous piezoelectric metal sheet; 15. Piezoelectric ceramic ring; 141. Microporous area; 2. Carrier gas transmission channel; 21. Carrier gas pipeline; 22. Compensating gas pipeline; 3. Aerosol transmission unit; 4. Sealing fixed ring; 5. Microporous piezoelectric single cell sampling device; 6. ICP matrix tube; 7. ICP source; 8. Quadrupole mass analyzer; 9. Ion detector. DETAILED DESCRIPTION

[0032] In order to make the purpose, technical solution and advantages of the present invention clearer, embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements with the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0033] refer to Figure 1The microporous piezoelectric single cell sampling device provided by the present invention is used in the sample introduction system of the ICP-MS analyzer to introduce the atomized sample into the ICP matrix tube. The microporous piezoelectric single cell sampling device 5 includes a sample processing part 1, a carrier gas transmission channel 2 and an aerosol transmission part 3. The sample processing part 1 is coaxially arranged at one end of the aerosol transmission part 3, and the carrier gas transmission channel 2 is connected to the aerosol transmission part 3; the aerosol transmission part 3 is conical; the sample processing part 1 includes a microporous piezoelectric cell screening component 11, a microporous piezoelectric cell The introduction component 12 and the linkage circuit controller 13, the microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12 are arranged at intervals, and are sealed at the inlet end of the aerosol transmission part 3 through the sealing fixing ring 4, and the linkage circuit controller 13 controls the piezoelectric effect of the microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12; the carrier gas transmission channel 2 is arranged along the inner wall of the aerosol transmission part 3 to generate a vortex carrier gas flow in the fog chamber cavity; the outlet end of the aerosol transmission part 3 is connected to the ICP matrix tube. The trace cell solution is controlled by a pipette or a syringe pump to be loaded into the dorsal central area of ​​the microporous piezoelectric cell screening component driven by a low-frequency AC signal, and the linkage circuit controller 13 controls the microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12 at the same time. Under the action of piezoelectric effect, the microporous piezoelectric cell screening component 11 converts the cell solution from the back side into aerosol, and the cells with a particle size larger than the micropore size are retained in the residual solution on the back side, and the cells with a particle size smaller than the micropore size are transmitted with the aerosol to the back center area of ​​the microporous piezoelectric cell introduction component 12. The cell solution after particle size screening is again converted into an aerosol containing cell samples by the microporous piezoelectric cell introduction component 12 through the piezoelectric effect. The aerosol is introduced into the ICP along the central axis direction of the aerosol transmission part 3 under the guidance of the vortex carrier gas to complete the processes of desolvation, atomization, excitation and ionization, and the final signal is detected by a mass spectrometer.

[0034] In some embodiments, Figure 2a As shown, the microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12 are formed by bonding a microporous piezoelectric metal sheet 14 with a microporous structure in the center and a piezoelectric ceramic ring 15. The micropores of the microporous piezoelectric metal sheet 14 are formed into a rounded structure by 3D printing, which avoids the damage to the cells caused by the micropore edge burrs produced by conventional micropore processing. Figure 2b As shown, the process of cells entering the microporous area 141 is demonstrated.

[0035] In some embodiments, the microporous piezoelectric cell screening component 11 , the microporous piezoelectric cell introducing component 12 and the aerosol transmission part 3 are coaxially placed and tightly connected to each other via a sealing fixing ring 4 .

[0036] In some embodiments, the microporous piezoelectric metal sheet 14 of the microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12 has a diameter of 3 to 50 mm, and can be made of platinum, silver, gold, stainless steel, tantalum, tungsten, and other metal materials, with several to tens of thousands of micropores in the central area. The microporous piezoelectric metal sheet 14 is designed and processed using 3D printing, and the microporous channel outlet and inlet are designed with rounded corners to ensure the integrity of the cells during transmission. The inlet diameter of the micropore is 50 to 500 μm, and the diameter of the micropore spray port is 10 to 100 μm. The inner diameter of the piezoelectric ceramic ring 15 can be selected from 2.0 to 40.0 mm, and the outer diameter can be selected from 3.0 to 50.0 mm, generally made of materials such as lead zirconate titanate (PZT) or alumina.

[0037] In some embodiments, the linkage circuit controller 13 can output through a signal generator of any brand and model, or through a homemade integrated circuit board, and the output frequency of the microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12 is 10 to 1000 kHz, the voltage is 1 to 30 V, and the output current is 0.001 to 1A.

[0038] In some embodiments, the cavity of the aerosol transmission part 3 is streamlined, the outside of the pipeline connected to the microporous piezoelectric cell introduction component 11 is designed with a threaded structure, the carrier gas pipeline 21 and the compensation gas pipeline 22 are tangent to the inner wall of the aerosol transmission part 3, and a vortex carrier gas flow can be generated in the mist chamber cavity. The total cavity volume is 10-200mL, and the material can be selected from rubber, resin, glass or quartz and other materials.

[0039] In some embodiments, the size of the sealing fixing ring 4 connecting the microporous piezoelectric cell screening component 11, the microporous piezoelectric cell introduction component 12 and the aerosol transmission part 3 matches the size of the microporous piezoelectric cell screening component 11, the microporous piezoelectric cell introduction component 12 and the aerosol transmission part 3, and the material can be an elastic material such as rubber or resin. The microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12 are coaxially placed with a center spacing of 5 to 30 mm. The carrier gas flow rate range is 0.1 to 2.0 Lmin -1 , the compensation gas flow rate range is 0~2.0L min -1 The atomization rates of the microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12 are both between 1 and 30 μL min -1 .

[0040] In some embodiments, Figure 3 As shown, the microporous piezoelectric single-cell sampling device for ICP-MS analysis proposed by the present invention can be used in conjunction with an inductively coupled plasma mass spectrometer.

[0041] The microporous piezoelectric single cell sampling device provided by the present invention is described in detail below in conjunction with specific embodiments.

[0042] The metal sheet of the microporous piezoelectric atomization device in the embodiment is printed in an integrated manner by the submicron resolution CERES micro-nano metal 3D printing system based on cathode electrochemical deposition technology at 30°C in an additive manner, and the metal sheet of the piezoelectric effect droplet injection device with micropores in the center is formed by connecting the materials layer by layer. Microfluidics adjusts the pulse gas pressure to transfer the metal electrolyte containing 0.2 mol / L Pt ions to the printing tip of the micro-nano metal 3D printing system through a 1μm diameter micro-channel. The Pt ions are at a current density of 2mA / cm at the moment the ion solution leaves the tip. 2 Under the action of the electrolysis process, pixel-level metal Pt solid is quickly crystallized, and patterned pixel stacking is achieved through the movement of the ion probe and the platform. The printing speed is fixed at 40μm / s, and finally a metal sheet with a micron-level smooth microporous structure is formed.

[0043] The printed microporous piezoelectric metal sheet is a platinum sheet with an outer diameter of 16 mm, and the piezoelectric ceramic ring has an outer diameter of 16 mm and an inner diameter of 10 mm. The cell solution is added to the center area of ​​the metal sheet of the microporous piezoelectric cell screening component through a syringe pump to introduce the sample. The operating frequency of the microporous piezoelectric cell screening component and the microporous piezoelectric cell introduction component are both 108 kHz, the voltage is 10 V, the current is 0.005 A, and the injection rate is 10 μL min -1 ; The inductively coupled plasma frequency is 27.12MHz, the power is 1300W, the ICP sampling depth is 8mm, and the mass spectrometer type is a quadrupole mass spectrometer.

[0044] The circuit diagram of the linkage circuit controller in the embodiment is as follows: Figure 7As shown in the figure, the circuit is designed by combining NE555 and MOS modules. The pin assignment of NE555 integrated circuit includes: (1): ground; (2): trigger pin; (3): output pin (4): reset pin (usually connected to high level to avoid false triggering); (5): control voltage (6): threshold pin (7): discharge pin (8): 5V power supply pin. By connecting two resistors (R1 and h1) and two capacitors (C1 and C2) to form a feedback loop, the trigger pin can apply a pulse to change the output state, and a square wave pulse signal of a certain frequency can be generated in the bistable mode. By adjusting the resistance value of h1, the frequency of the square wave pulse signal can be continuously adjusted from 1kHz to 1MHz. The output pin (pin 3) of NE555 is connected to the gate of MOS. The source and drain of MOS are connected to the negative pole of the power supply (GND) and one end of the micro inductor L respectively, and the other end of the inductor L is connected to the positive pole of the power supply (VCC). By controlling the gate voltage to adjust the current between the source and the drain, the electrical signal output by NE555 is amplified, and the amplification factor, that is, the final output electrical signal strength, can be further controlled by adjusting the H2 resistor. The microporous piezoelectric cell screening component (atomizer 1) and the microporous piezoelectric cell introduction component (atomizer 2) serve as terminal loads, and both control the circuit on and off through a button switch.

[0045] Example 1

[0046] The present invention provides a microporous piezoelectric single cell sampling device, the structure of which is as follows Figure 1 As shown in Figure 2.

[0047] The microporous piezoelectric metal sheets of the microporous piezoelectric cell screening component and the microporous piezoelectric cell introduction component have the same specifications: a total of 1000 micropores in the central diameter area of ​​5mm, a micropore entrance diameter of 60μm, a micropore spray port diameter of 20μm, and a micropore spacing of 150μm. The microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12 are connected to the aerosol transmission part 3 through an elastic sealing fixing ring 4, and the carrier gas pipeline 21, the compensation gas pipeline 22 and the aerosol transmission part 3 of the aerosol transmission part 3 are formed by quartz processing and welding. Under the action of the tangential vortex carrier gas, an axial central transmission channel is provided for the cell aerosol micelles, so that 100% of them enter the ICP for analysis and detection.

[0048] Experimental specific steps: 1) The number concentration is 10 10 The blue cell solution was diluted 10 5 2) Ignite the ICP plasma and adjust the mass spectrometer detector to single cell analysis mode. 3) The shaken blue cell solution is pumped through a syringe pump at a rate of 10 μL min -1) The aerosol is transported to the center of the microporous piezoelectric device at a uniform speed; 4) The electrical signal input of the microporous piezoelectric device is turned on, and under the action of the tangential eddy current carrier gas, all the blue cells carried by the aerosol are introduced into the ICP; 5) The original signal spectrum saved in the single cell analysis mode is output, and the blue cells are used 44 The Ca signal records the number of single particle events. By comparing the original signal spectrum obtained with the conventional pneumatic nebulization injection system (see Figure 4a and Figure 4b ), which verifies that the sample introduction efficiency of the micro-cell efficient introduction system based on piezoelectric effect droplet injection is 10-20 times higher than that of conventional pneumatic atomization, which helps to improve the throughput of cell analysis based on mass spectrometry.

[0049] Example 2

[0050] The microporous piezoelectric single cell sampling device of the present invention can be used for online screening of small-size cells in mixed cell samples for analysis. The device structure of this embodiment is the same as that of embodiment 1. The difference from embodiment 1 is that the microporous piezoelectric metal sheets of the microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12 have different specifications: the microporous piezoelectric metal sheet of the microporous piezoelectric cell screening component 11 has a total of 300 micropores in the central area, the micropore inlet diameter is 200μm, the micropore spray port diameter is 100μm, and the micropore spacing is 230μm; the microporous piezoelectric metal sheet of the microporous piezoelectric cell introduction component 12 has a total of 10 micropores in the central area, the micropore inlet diameter is 60μm, the micropore spray port diameter is 20μm, and the micropore spacing is 150μm. The cell sample type is a mixed solution of red blood cells and epithelial cells, and the number of the two types of cells is 10 respectively. 5 and 10 3 Pieces / mL.

[0051] The specific steps of the experiment are as follows: 1) Load 10μL of the cell mixture solution into the center area of ​​the microporous piezoelectric cell screening module through a pipette; 2) Turn on the electrical signal input of the microporous piezoelectric cell screening module, and the epithelial cells with larger particle sizes are intercepted by the microporous piezoelectric cell screening module, and the red blood cells with smaller particle sizes are transported to the center area of ​​the microporous piezoelectric cell introduction module with the aerosol to re-form droplets. 3) Turn on the electrical signal input of the microporous piezoelectric cell introduction module, and all the red blood cells carried by the aerosol are introduced into the ICP; 4) Output the original signal spectrum saved in the single cell analysis mode, and use 57 The Fe signal records the number of single particle events (e.g. Figure 5 5) At the same time, when the ICP is turned off, the aerosol beam of step 3) is collected offline using a slide, and the number of red blood cells and epithelial cells is observed under a microscope (as shown in Figure 6 As shown), it can be verified that the micro-cell efficient introduction system based on piezoelectric effect droplet injection can effectively retain epithelial cells and can introduce red blood cells in the solution with an efficiency of up to 96%.

[0052] For matters not mentioned above, the prior art applies.

[0053] Although some specific embodiments of the present invention have been described in detail through examples, those skilled in the art should understand that the above examples are for illustration only and are not intended to limit the scope of the present invention. Those skilled in the art to which the present invention belongs may make modifications or supplements to the described specific embodiments or replace them in a similar manner, but they will not deviate from the direction of the present invention or exceed the scope defined by the attached claims. Those skilled in the art should understand that any modifications, equivalent substitutions, improvements, etc. made to the above implementations based on the technical essence of the present invention should be included in the protection scope of the present invention.

Claims

1. A microporous piezoelectric single cell sampling device, used in the sample introduction system of an ICP-MS analyzer, to introduce atomized samples into an ICP matrix tube, characterized in that: The microporous piezoelectric single-cell sampling device (5) comprises a sample processing part (1), a carrier gas transmission channel (2) and an aerosol transmission part (3), wherein the sample processing part (1) is coaxially arranged at one end of the aerosol transmission part (3), and the carrier gas transmission channel (2) is connected to the aerosol transmission part (3); the aerosol transmission part (3) is conical; The sample processing part (1) comprises a microporous piezoelectric cell screening component (11), a microporous piezoelectric cell introduction component (12) and a linkage circuit controller (13); the microporous piezoelectric cell screening component (11) and the microporous piezoelectric cell introduction component (12) are arranged at intervals and are sealed at the inlet end of the aerosol transmission part (3) through a sealing fixing ring (4); and the linkage circuit controller (13) controls the piezoelectric effect of the microporous piezoelectric cell screening component (11) and the microporous piezoelectric cell introduction component (12); The carrier gas transmission channel (2) is arranged along the inner wall of the aerosol transmission part (3) to generate a vortex carrier gas flow in the spray chamber cavity; The outlet end of the aerosol transmission part (3) is in communication with the ICP matrix tube.

2. The microporous piezoelectric single cell sampling device according to claim 1, characterized in that: The microporous piezoelectric cell screening component (11) and the microporous piezoelectric cell introduction component (12) are both formed by bonding a microporous piezoelectric metal sheet (14) and a piezoelectric ceramic ring (15), and a microporous area (141) is arranged at the center of the microporous piezoelectric metal sheet (14).

3. The microporous piezoelectric single cell sampling device as claimed in claim 2, characterized in that: The distance between the microporous piezoelectric cell screening component (11) and the microporous piezoelectric cell introduction component (12) is 5 to 30 mm.

4. The microporous piezoelectric single cell sampling device as claimed in claim 2, characterized in that: The diameter of the microporous piezoelectric metal sheet ranges from 3 to 50 mm.

5. The microporous piezoelectric single cell sampling device as claimed in claim 3, characterized in that: The inlet diameter of the micropores on the microporous piezoelectric metal sheet (14) ranges from 50 to 500 μm.

6. The microporous piezoelectric single cell sampling device as claimed in claim 3, characterized in that: The diameter of the spray outlet of the micropores on the microporous piezoelectric metal sheet is in the range of 10 to 100 μm.

7. The microporous piezoelectric single cell sampling device as claimed in claim 2, characterized in that: The carrier gas transmission channel (2) comprises a carrier gas pipeline (21) and a compensation gas pipeline (22), and the carrier gas pipeline (21) and the compensation gas pipeline (22) are arranged radially symmetrically on the inner wall of the aerosol transmission part (3) along the microporous piezoelectric metal sheet (14).

8. The microporous piezoelectric single cell sampling device as claimed in claim 1, characterized in that: The cavity volume of the aerosol transmission part (3) is between 10 and 200 mL.

9. The microporous piezoelectric single cell sampling device as claimed in claim 2, characterized in that: The microporous piezoelectric cell screening component (11) and the microporous piezoelectric cell introduction component (12) are integrally formed by 3D printing.

10. An inductively coupled plasma mass spectrometer comprising the microporous piezoelectric single cell sampling device according to any one of claims 1 to 9.

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