A microporous piezoelectric single cell sampling device and its application

Abstract

The present invention provides a microporous piezoelectric single-cell sampling device and its application, which belongs to the field of single-cell detection technology. The microporous piezoelectric single-cell sampling device includes a sample processing part, a carrier gas transmission channel and an aerosol transmission part. The sample processing part includes a microporous piezoelectric cell screening component, a microporous piezoelectric cell introduction component and a linkage circuit controller. 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; 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 through airflow regulation; the device avoids the generation of waste liquid and effectively reduces pollution to the environment. It can also be used in conjunction 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 microporous piezoelectric single cell sampling system and a single cell detection method. Background Art

[0002] Single-cell analysis has become a significant research area in recent years. It can reveal individual cell characteristics, facilitate the study of cellular complexity and intercellular variability, and possesses significant biological value. Inductively coupled plasma mass spectrometry (ICP-MS), with its exceptional ability to analyze trace metal elements, has played a key role in determining the content and distribution of metal elements within single cells. In recent years, metal nanoparticles, particularly functionalized metal nanoparticles, have been widely used in bioimaging, intracellular sensing, and disease treatment due to their unique chemical and physical properties. However, little has been reported on their intracellular accumulation, distribution, and heterogeneity. In biomedical applications and toxicological analysis, the study of nanoparticle cellular uptake plays a crucial role. Therefore, developing high-throughput, high-precision single-cell analysis methods to investigate the effects of nanoparticles on cells or organisms is of vital importance. However, conventional sample introduction systems based on pneumatic atomization typically have an injection efficiency of 1–10%, meaning that a significant portion of the sample is lost in the wastewater. Inefficient sample introduction systems are not conducive to analyzing precious, trace cell samples, and the resulting wastewater also poses a risk of environmental contamination. Therefore, an efficient sample injection system is very important for ICP-MS analysis instruments.

[0003] Several high-efficiency sample introduction systems have been reported, including invention patents US10497550(B1) and CN207816913U, which disclose membrane desolvation devices (MDs); CN104931420B, which discloses an ultrasonic nebulization (USN) sample introduction system; and CN217954234U and CN207233698U, which disclose electrothermal vaporization (ETV) sample introduction systems. High-efficiency sample introduction systems are a significant obstacle to the development of large-scale inorganic analytical instruments. While these reported systems can improve sample introduction efficiency and enhance analytical sensitivity, they each have drawbacks. MDs are expensive (tens of thousands of dollars), residue cleaning is difficult, and maintenance is cumbersome. USNs also suffer from high costs and residue cleaning issues. Furthermore, existing commercial USNs have high sample introduction rates, which cannot meet the needs of trace sample analysis. ETV utilizes thermal energy to convert analytes into gaseous species, providing a certain degree of sample selectivity. However, its analytical throughput is low, and it is not suitable for the introduction of nanoparticles and cell-based samples. Therefore, it has not yet become a routine sample introduction device for ICP-MS. Therefore, it is highly desirable to develop a new, efficient introduction system for trace samples with high throughput, high injection efficiency, and low carryover.

[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, making it a very effective atomization sampling method. Compared with conventional pneumatic atomization sampling systems, this method has lower costs and can produce easily controllable low-speed aerosol micelles, 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 for the following reasons: (1) Cells vary in size, and the micropore 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 and cause blockage; (2) The micropore edges of existing commercial microporous piezoelectric atomization sheets have irregular sharp edges, which can easily damage cells in the process of passing through the micropores. Therefore, the present invention proposes to develop a new type of microporous piezoelectric cell atomization device for efficient cell biological sample introduction system for ICP-MS analysis. Summary of the Invention

[0005] The purpose of the present invention is to address the above-mentioned shortcomings of the existing technology and provide a microporous piezoelectric single-cell sampling device and application, which can screen cells of different sizes online according to cell size, and achieve efficient introduction of specific target cells. The device has extremely high transmission efficiency and greatly reduces the sample amount 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 for use in a 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 portion, a carrier gas transmission channel, and an aerosol transmission portion. The sample processing portion is coaxially arranged at one end of the aerosol transmission portion, and the carrier gas transmission channel is connected to the aerosol transmission portion. The aerosol transmission portion is tapered.

[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 spaced apart and 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 mist 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 provided in the center of the microporous piezoelectric metal sheet.

[0010] Furthermore, the distance between the microporous piezoelectric cell screening component and the microporous piezoelectric cell introduction 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] A 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 sampling systems and other high-efficiency sampling systems, the microporous piezoelectric single-cell sampling device provided by the present invention is as follows: the core component of the device provided by the present invention is made of a microporous piezoelectric atomization sheet by 3D printing, and the rounded microporous structure made thereby 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 microporous 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 by 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 This is a schematic structural diagram of the microporous piezoelectric single-cell sampling device provided by the present invention;

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

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

[0025] Figure 3 Schematic diagram of the structure 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 microporous 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 ions detected in each 10 millisecond time window. 44 The intensity of the Ca ion counting signal is expressed as 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 intensity of the Ca ion counting signal is expressed as 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 The horizontal axis represents the monitoring time, and the vertical axis represents the ion signal of Fe detected in each 10 millisecond time window. 57 The Fe ion counting signal intensity is expressed as pulse peaks, and the number of pulse peaks corresponds to the number of red blood cells detected in that time period;

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

[0030] Figure 7 1 is a circuit diagram controlled by a 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 torque tube; 7. ICP source; 8. Quadrupole mass analyzer; 9. Ion detector. DETAILED DESCRIPTION

[0032] To make the objectives, technical solutions, and advantages of the present invention more apparent, embodiments of the present invention are described in detail below. 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 having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended only to explain the present invention and are not to be construed 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 are spaced apart from the microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12. The microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12 are sealed at the inlet of the aerosol transmission section 3 via a sealing retaining ring 4. 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 section 3 to generate a vortex carrier gas flow in the mist chamber cavity. The outlet of the aerosol transmission section 3 is connected to the ICP matrix tube. A small amount of cell solution is loaded into the dorsal center area of ​​the microporous piezoelectric cell screening component driven by a low-frequency AC signal via a pipette or syringe pump. The linkage circuit controller 13 simultaneously controls the microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12. Under the action of the piezoelectric effect, the microporous piezoelectric cell screening component 11 converts the cell solution drawn in from the back into an aerosol. Cells with particle sizes larger than the micropore diameter are retained in the residual solution on the back, while cells with particle sizes smaller than the micropore diameter are transported with the aerosol to the back center area of ​​the microporous piezoelectric cell introduction component 12. After particle size screening, the cell solution is again converted by the piezoelectric effect by the microporous piezoelectric cell introduction component 12 into an aerosol entrained with the cell sample. Guided by the vortex carrier gas, the aerosol is introduced into the ICP along the central axis of the aerosol transmission unit 3 to complete the desolvation, atomization, excitation, and ionization processes. The resulting signal is detected by a mass spectrometer.

[0034] In some embodiments, as 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 corner 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 diameter of the micropore entrance 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, and is 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, with an output frequency of 10 to 1000 kHz, a voltage of 1 to 30 V, and an output current of 0.001 to 1 A for the microporous piezoelectric cell screening component 11 and the microporous piezoelectric cell introduction component 12.

[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 12 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.

[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 distance of 5 to 30 mm. The carrier gas flow rate range is 0.1 to 2.0 L min -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 in the range of 1 to 30 μL min -1 .

[0040] In some embodiments, as 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 with reference to specific embodiments.

[0042] In the embodiment, the metal sheet of the microporous piezoelectric atomization device is printed integrally by the CERES micro-nanometal 3D printing system with submicron resolution based on cathode electrochemical deposition technology at 30°C in an additive manner, and the metal sheet of the piezoelectric effect droplet ejection device with micropores in the center is formed by connecting the materials layer by layer. Microfluidics adjusts the pulse gas pressure to transport the metal electrolyte containing 0.2 mol / L Pt ions through a 1 μm diameter micro-pipeline to the printing tip of the micro-nanometal 3D printing system. The Pt ions are discharged at a current density of 2 mA / 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 generated. 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 via 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 27.12 MHz, the power is 1300 W, the ICP sampling depth is 8 mm, 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 be applied with 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 drain electrodes, the NE555's output signal is amplified. The amplification factor, or the final output signal strength, can be further controlled by adjusting the H2 resistor. The microporous piezoelectric cell screening assembly (atomizer 1) and microporous piezoelectric cell introduction assembly (atomizer 2) serve as terminal loads, both of which are controlled by a pushbutton switch to turn the circuit on and off.

[0045] Example 1

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

[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, the inlet diameter of the micropores is 60μm, the spray port diameter of the micropores is 20μm, and the micropore spacing is 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. The carrier gas pipeline 21, the compensation gas pipeline 22 and the aerosol transmission part 3 of the aerosol transmission part 3 are composed of quartz processing and welding. Under the action of the tangential eddy current 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 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 injected into the syringe pump at a rate of 10 μL min -1The 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. The original signal spectrum obtained by the conventional pneumatic nebulization injection system is compared (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-sized cells in mixed cell samples for analysis. The structure of the device in this embodiment is the same as that in 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] Specific experimental steps: 1) Load 10μL of 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 size are intercepted by the microporous piezoelectric cell screening module, and the red blood cells with smaller particle size are transported with the aerosol to the center area of ​​the microporous piezoelectric cell introduction module 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 Fe signal records the number of single particle events (such as Figure 5 5) At the same time, when the ICP is turned off, use a glass slide to collect the aerosol beam in step 3) offline, and observe the number of red blood cells and epithelial cells under a microscope (as shown). 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] Any matters not mentioned above shall be subject to the existing technology.

[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 of the present invention 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 appended claims. Those skilled in the art should understand that any modifications, equivalent substitutions, improvements, etc. made to the above embodiments based on the technical essence of the present invention should be included in the scope of protection 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 tapered; 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 spaced apart and 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 communicated 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 adhering a microporous piezoelectric metal sheet (14) and a piezoelectric ceramic ring (15), and a microporous area (141) is provided at the center of the microporous piezoelectric metal sheet (14).

3. The microporous piezoelectric single cell sampling device according to 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 according to claim 2, wherein: The diameter of the microporous piezoelectric metal sheet ranges from 3 to 50 mm.

5. The microporous piezoelectric single cell sampling device according to claim 3, wherein: The entrance 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 according to claim 3, wherein: The diameter of the spray port of the micropores on the microporous piezoelectric metal sheet ranges from 10 to 100 μm.

7. The microporous piezoelectric single cell sampling device according to claim 2, wherein: 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 according to claim 1, wherein: The cavity volume of the aerosol transmission part (3) is 10 to 200 mL.

9. The microporous piezoelectric single cell sampling device according to claim 2, wherein: 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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