Rapid enrichment device based on bulk acoustic waves and working method

Abstract

The invention discloses a rapid enrichment device and method based on bulk acoustic waves, and belongs to the technical field of acoustic wave device application. The device comprises an enrichment assembly, a control assembly and a lifting device, the two piezoelectric ceramic pieces are oppositely arranged, the distance between the two piezoelectric ceramic pieces is integral multiple of the wavelength of sound waves generated by the piezoelectric ceramic pieces, standing waves are formed in a medium solution in a convex structure, biological particles in a centrifugal tube are fixed to sound wave nodes, and then enrichment is achieved by lifting the centrifugal tube. A cooling platform is arranged at the bottom of the device shell, a temperature sensor is arranged in the convex device shell, and the temperature of a medium solution is regulated at any time. The device has the advantages of low damage, operation flexibility and efficient enrichment, and has wide application prospects in the fields of biomedicine, clinical detection, drug development and the like.

Description

Technical Field

[0001] The invention relates to the technical field of acoustic wave device applications, and in particular to a device and method for rapidly enriching biological particles based on bulk acoustic waves. Background Art

[0002] With the continuous development of biomedicine, drug development and basic research, cell enrichment technology has gradually become one of the key tools in research and clinical applications. Rapid enrichment and analysis of rare cell populations such as circulating tumor cells can provide important basis for early diagnosis of diseases. For example, circulating tumor cell enrichment technology in liquid biopsy can achieve non-invasive early detection and monitoring of cancer. At present, the traditional sample pretreatment process relies on large equipment such as ultracentrifuges. This process is not only cumbersome and time-consuming, but also has high instrument and equipment costs and high experimental costs. Due to the structural problems of the centrifuge tube in the centrifuge, the particle clusters enriched by the centrifuge will be close to the bottom of the centrifuge tube. This solution will increase the difficulty of extracting the target particles, and also cause mechanical damage to the fragile and expensive target biological particles, which is not conducive to subsequent analysis and detection. Acoustic tweezers technology acts on cells in a non-contact manner, without the need for chemical reagents or physical invasion, and can enrich or sort cells without destroying cell activity. Surface acoustic wave technology can achieve rapid enrichment of multi-scale biological particles, but its processing flux upper limit is low, and its microchannel production time cost is large, and its processing efficiency is low when processing multiple large-volume samples. In addition, surface acoustic wave enrichment technology usually requires the assistance of devices such as microinjection pumps, and the overall system is highly complex and the cost of instruments and equipment is relatively high. Summary of the invention

[0003] In order to solve the problems existing in the prior art, the present invention aims to provide a fast, simple, visual large-capacity biological particle enrichment device, which uses bulk acoustic wave technology to quickly enrich the target biological particles in the sample solution in the centrifuge tube in a fast and visual manner at the bottom of the centrifuge tube, facilitating the subsequent extraction of biological particle samples. This technology can be used in subsequent biological analysis and medical diagnosis and other fields.

[0004] The technical solution adopted to achieve the above purpose is: a rapid enrichment device based on bulk acoustic wave, the device includes a control component, an enrichment component and a lifting device; the control component includes a signal generation module, a temperature control module and a lifting device speed adjustment knob;

[0005] The enrichment component of the device adopts a convex structure containing a medium solution arranged on a cooling platform, the convex structure adopts a rectangular structure surrounded by two shell side plates and two transparent end plates on the shell bottom plate, and a shell top plate is arranged above the rectangular structure;

[0006] Piezoelectric ceramic sheets are respectively arranged at the relative positions of the two side plates of the housing, and the distance between the two piezoelectric ceramic sheets is an integral multiple of the wavelength of the sound wave; the positive and negative electrodes of the piezoelectric ceramic sheets on both sides are respectively connected to the signal generating module;

[0007] The top plate of the shell is provided with a centrifuge tube placement hole slot and a temperature sensing hole for installing a temperature sensor, the upper end of the centrifuge tube is clamped in the centrifuge tube clamping slot, the centrifuge tube clamping slot is placed on the top plate of the shell, and the bottom of the centrifuge tube is inserted into the medium solution of the convex structure; the centrifuge tube is placed between two piezoelectric ceramic sheets; a centrifuge tube clamping slot rack for lifting is provided above the centrifuge tube clamping slot;

[0008] An internal drive motor is arranged in the base of the lifting device, and the internal drive motor drives the lifting rod to move up and down, and a lifting slot matching with the centrifuge tube slot frame is arranged on the lifting rod;

[0009] The cooling platform, the temperature sensor and the temperature control module in the enrichment component are electrically connected, and the lifting device is electrically connected to the lifting device speed adjustment knob, and the lifting device speed adjustment knob is used to control the speed of the lifting rod.

[0010] The piezoelectric ceramic piece is circular, rectangular or elliptical.

[0011] The diameter of the centrifuge tube is smaller than the distance of the piezoelectric ceramic sheet in the length direction of the shell side plate, and the bottom end of the centrifuge tube is higher than the bottom end of the piezoelectric ceramic sheet; the centrifuge tube is in the area covered by the body acoustic wave vibration.

[0012] The top end of the piezoelectric ceramic sheet is tangent to or in contact with the bottom surface of the top plate of the housing.

[0013] The medium solution is water, or a glycerol aqueous solution with a glycerol volume fraction of 10%-30%.

[0014] The piezoelectric ceramic sheet is provided with a piezoelectric ceramic sheet waterproof layer, a piezoelectric ceramic sheet negative electrode and a piezoelectric ceramic sheet positive electrode in sequence from the inside to the outside of the convex structure, and the distance between the piezoelectric ceramic sheet waterproof layers on both sides is an integral multiple of the wavelength of the sound wave.

[0015] The start temperature of the cooling platform is 27°C and the stop temperature is 23°C.

[0016] A working method of a rapid enrichment device based on bulk acoustic waves comprises the following steps:

[0017] S1. Inject the medium solution into the convex structure until the liquid level exceeds the highest point of the piezoelectric ceramic piece; place the temperature sensor in the temperature sensing hole of the top plate of the shell, monitor the temperature of the medium solution at any time, and adjust the cooling platform;

[0018] S2. Turn on the cooling platform and set its temperature range to 23-27°C. When the medium solution temperature exceeds 27°C, the temperature control module controls the cooling platform to start working, and stops working when the medium solution temperature drops to 23°C.

[0019] S3, adjusting the signal generating module to adjust the input frequency and amplitude of the RF signal;

[0020] S4. Put the biological particle sample into a centrifuge tube, put the centrifuge tube into the centrifuge tube slot, place the centrifuge tube slot rack into the lifting slot, lower the centrifuge tube into the convex structure, and adjust the liquid level inside the centrifuge tube to make it horizontal;

[0021] And set the initial lifting speed of the lifting rod in the lifting device through the lifting device speed adjustment knob;

[0022] S5, turn on the signal generating module output, observe the real-time situation inside the centrifuge tube through the transparent end plate, and after it forms a stable line arrangement, turn on the lifting device, and the internal driving motor drives the lifting rod to move upward, and the lifting rod further drives the centrifuge tube slot rack and the centrifuge tube slot to finally drive the centrifuge tube to move upward;

[0023] S6. Observe the lifting situation in real time during the lifting process. When the centrifuge tube leaves three quarters of the working area of ​​the piezoelectric ceramic sheet, adjust the lifting speed to half of the initial lifting speed and wait for the lifting to be completed.

[0024] S7. When the bottom of the centrifuge tube is lifted above the sonic working area, the lifting device is closed to complete the enrichment. After the enrichment is completed, the biological particles are stored at the bottom of the centrifuge tube.

[0025] The biological particle sample is a liquid sample containing cells, organelles or pathogens.

[0026] The cell-containing liquid sample is human or animal body fluid or effusion, cell culture fluid, strain culture fluid or tissue culture fluid.

[0027] Furthermore, a working method of a rapid enrichment device based on bulk acoustic waves comprises the following steps:

[0028] S1. Inject the medium solution into the convex structure until the liquid level exceeds the highest point of the piezoelectric ceramic piece; place the temperature sensor in the temperature sensing hole of the top plate of the shell, monitor the temperature of the medium solution at any time, and adjust the cooling platform;

[0029] S2. Turn on the cooling platform and set its temperature range to 23-27°C. When the medium solution temperature exceeds 27°C, the cooling platform starts to work and stops working when the medium solution temperature drops to 23°C.

[0030] S3, adjust the signal generation module, adjust the input frequency and amplitude of the RF signal; set the input frequency to 1MHz-100MHz, connect an oscilloscope to both ends of the piezoelectric ceramic, adjust the input voltage after amplification by the power amplifier to 80V, and turn off the input;

[0031] S4. Load the biological particle sample into a centrifuge tube, place the centrifuge tube into the centrifuge tube slot, lower the centrifuge tube into the convex structure, and adjust the liquid level inside the centrifuge tube to make it horizontal; turn on the rapid lifting device; set the lifting speed of the rapid lifting device to 1mm / s, and set the lifting mode of the rapid lifting device to automatic lifting.

[0032] S5. Turn on the output of the signal generating module, observe the real-time situation inside the centrifuge tube through the transparent end plate, and turn on the rapid lifting device after a stable line arrangement is formed;

[0033] S6. Observe the lifting situation in real time during the lifting process. When the centrifuge tube leaves three quarters of the working area of ​​the piezoelectric ceramic sheet, adjust the lifting speed to 0.5 mm / s and wait for the lifting to be completed.

[0034] S7. When the bottom of the centrifuge tube is lifted above the sonic working area, the lifting device is closed to complete the enrichment. After the enrichment is completed, the biological particles are stored at the bottom of the centrifuge tube.

[0035] The convex structure is provided with a piezoelectric ceramic slot, a centrifuge tube placement hole slot and a slot for observation, and a shell for increasing the contact area with the cooling platform is provided at the bottom. The piezoelectric ceramic slot is provided with a first piezoelectric ceramic and a second piezoelectric ceramic, respectively, for generating standing waves and forming standing wave nodes, so that the bulk acoustic wave propagates in the internal medium solution into the centrifuge tube, driving the biological particles in the centrifuge tube to move to the standing wave nodes for subsequent enrichment. The lifting device includes an internal drive motor, a lifting rod and a lifting slot, which are used to lift the centrifuge tube to complete the enrichment. The frequency of the piezoelectric ceramic is 1.7MHz, and the shape of the piezoelectric ceramic is round with a diameter of 20mm.

[0036] The size of the centrifuge tube slot and the diameter of the piezoelectric ceramic piece can be adjusted according to the diameter of the centrifuge tube; the diameter of the centrifuge tube placed in the centrifuge tube slot should be smaller than the diameter of the piezoelectric ceramic piece.

[0037] The cooling platform is used to control the temperature of the solution inside the convex device shell so as to keep the temperature inside the convex device shell constant.

[0038] The signal generating module is used to output the radio frequency signal to drive the piezoelectric ceramic. The signal amplifying device is used to amplify the radio frequency signal generated by the signal generating device, and then directly connect the piezoelectric ceramic. The speed adjusting knob of the lifting device is used to adjust the lifting speed at any time according to different biological particles, and to control the lifting speed at different positions of the centrifuge tube.

[0039] Compared with the prior art, the present invention has the following beneficial effects:

[0040] Two piezoelectric ceramic sheets are arranged opposite to each other, with a spacing equal to an integer multiple of the wavelength of the sound waves generated by the piezoelectric ceramic sheets, so that a standing wave is formed in the medium solution inside the convex structure, and the biological particles in the centrifuge tube are fixed on the sound wave nodes, and then enriched by lifting the centrifuge tube. A cooling platform is set at the bottom of the device shell, and a temperature sensor is set inside the convex device shell to adjust the temperature of the medium solution at any time.

[0041] (1) Compared with the traditional centrifuge for extracting biological particles, the present invention provides an enrichment device and method based on bulk acoustic wave. The device can complete the enrichment of biological particles in a centrifuge tube in a short time, and can directly extract the target biological particles at the bottom of the centrifuge tube without resuspending the biological particles, thereby improving the extraction efficiency.

[0042] (2) The enrichment method provided by the present invention is easy to operate. The entire enrichment process only requires turning on the signal and selecting the speed to complete the extraction of the target biological particles. This simplifies the operation during the biological particle processing process and reduces the damage to the biological particles caused by high-speed centrifugation, which is of significant significance for maintaining cell activity.

[0043] (3) The device provided by the present invention visualizes the enrichment process of biological particles in the centrifuge tube, and the enrichment process can be operated at any time. The time for increasing the sonication time can be selected according to the real-time situation, and the operability is high.

[0044] (4) The enrichment method provided by the present invention can be applied to the enrichment of biological particles of multiple scales, and can also be applied to the enrichment of centrifuge tubes with larger capacity, and has a wide range of applications.

[0045] (5) The device provided by the present invention acts on cells in a non-contact manner without the need for chemical reagents or physical damage, and can enrich cells without destroying cell activity.

[0046] Bulk acoustic wave enrichment uses the compression and stretching effects of sound waves to manipulate biological particles, which usually does not cause significant damage or stress to cells and is suitable for cell enrichment with high survival rate requirements. The propagation mode of bulk acoustic waves is relatively uniform. When performing cell operations in a larger volume of fluid, it can exert consistent acoustic radiation force on cells within the entire fluid volume, and can achieve a more uniform and efficient enrichment effect during large-scale cell sorting. Real-time monitoring and regulation can usually be achieved through a bulk acoustic wave-based enrichment system. Sensors are used to monitor cell behavior and adjust acoustic wave parameters as needed to optimize the enrichment effect and provide support for subsequent detection and analysis. Rapid cell enrichment technology plays an important role in basic scientific research and has broad application prospects in clinical diagnosis, drug development, cell therapy and other fields. It is of great significance to promote precision medicine and personalized treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] The accompanying drawings are intended to provide a further understanding of the present invention, and the accompanying drawings are incorporated into the specification and constitute a part of the specification. In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings required for use in the embodiments or the prior art description will be briefly introduced below.

[0048] Figure 1 It is a schematic diagram of the structure of a rapid enrichment device based on bulk acoustic waves.

[0049] Figure 2 It is a schematic diagram of the structure of the enrichment component.

[0050] Figure 3 It is an exploded diagram of the structure of the enrichment component.

[0051] Figure 4 It is a schematic diagram of the cross-sectional structure of a piezoelectric ceramic sheet.

[0052] Figure 5 It is a structural schematic diagram of the lifting device.

[0053] Figure 6 These are experimental effect diagrams of Hela cells before, during and after enrichment provided in an embodiment of the present invention.

[0054] In the figure: 1. piezoelectric ceramic sheet, 1a. negative electrode of piezoelectric ceramic sheet, 1b. positive electrode of piezoelectric ceramic sheet, 1c. waterproof layer of piezoelectric ceramic sheet, 2. side plate of outer shell, 3. cooling platform, 4. centrifuge tube slot, 5. transparent end plate, 6. hole slot for centrifuge tube, 7. centrifuge tube, 8. top plate of outer shell, 9. bottom plate of outer shell, 10. temperature sensing hole, 11. centrifuge tube slot rack, 12. lifting device, 12a. lifting rod, 12b. internal driving motor, 12c. device base, 12d. lifting slot, 13. signal generating module, 14. temperature control module, 15. lifting device speed regulating module. DETAILED DESCRIPTION

[0055] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in the field without making creative work are within the scope of protection of the present invention.

[0056] Figures 1 to 4 A rapid enrichment device based on bulk acoustic waves is shown, which includes a control component, an enrichment component and a lifting device 12; the control component includes a signal generating module 13, a temperature control module 14 and a lifting device speed adjustment knob 15.

[0057] In the lifting device 12, an internal drive motor 12b is arranged in the device base 12c, and the internal drive motor 12b drives the lifting rod 12a to move up and down. The lifting rod 12a is provided with a lifting slot 12d matched with the centrifuge tube slot frame 11. In the enrichment component, the cooling platform 3, the temperature sensor and the temperature control module 14 are electrically connected, and the lifting device 12 is electrically connected to the lifting device speed adjustment knob 15, and the lifting device speed adjustment knob 15 is used to control the speed of the lifting rod 12a. The cooling platform reduces the temperature change of the medium solution inside the device and reduces the change of the speed of the sound wave in the medium solution; the temperature of the cooling platform is adjusted at any time through the temperature sensor inside the convex device shell; the signal generating module is used to output the radio frequency signal to drive the piezoelectric ceramic sheet to work; the rapid lifting device is used to quickly lift the centrifuge tube after the sound wave fixes the Hela cells to achieve the enrichment effect.

[0058] Working mechanism of the lifting device: adjust the speed of the lifting device at 15, turn on the lifting device switch, start to lift slowly and steadily at 12a, and stop lifting after the centrifuge tube is completely out of the working area of ​​the piezoelectric ceramic sheet to complete the enrichment.

[0059] The lifting device 12 includes a lifting rod 12a and an internal driving motor 12b. The device bottom 12c is used to change the overall height of the lifting device. The lifting slot 12d is used to place the centrifuge tube slot and drive the movement of the centrifuge tube. The signal generating module 13 is used to provide signal drive to the piezoelectric ceramic sheet. The temperature control module 14 is used to adjust the starting working temperature and the stopping working temperature of the cooling platform. The lifting device speed adjustment knob 15 is used to adjust the lifting speed at any time.

[0060] The enrichment component adopts a convex structure with a medium solution set on the cooling platform 3. The convex structure adopts a rectangular structure surrounded by two shell side panels 2 and two transparent end panels 5 on the shell bottom plate 9, and a shell top plate 8 is set above the rectangular structure; two windows are set in the front and back of the convex structure respectively, using transparent end panels 5, for real-time observation of the enrichment situation inside the centrifuge tube. The bottom of the convex structure adopts a shell bottom plate with a larger contact area with the cooling platform, which is used to increase the contact area between the cooling platform and the medium solution and enhance the influence of the cooling platform on the medium solution; a temperature sensor hole is set on the shell top plate for placing a temperature sensor to adjust the temperature of the cooling platform.

[0061] Piezoelectric ceramic sheets 1 are respectively arranged at the opposite positions of the two side panels 2 of the housing. The piezoelectric ceramic sheets are placed opposite to each other on both sides of the centrifuge tube to form standing waves, and the biological particles in the centrifuge tube are linearly arranged at the nodes of the standing waves. The distance between the two piezoelectric ceramic sheets 1 is an integer multiple of the wavelength of the sound wave, which is convenient for forming standing waves; the positive and negative electrodes of the piezoelectric ceramic sheets on both sides are respectively connected to the signal generation module 13 of the control component;

[0062] The top plate 8 of the shell is provided with a centrifuge tube placement hole groove 6 and a temperature sensing hole 10 for installing a temperature sensor. The upper end of the centrifuge tube 7 is clamped in the centrifuge tube clamping groove 4, and the centrifuge tube clamping groove 4 is placed on the top plate 8 of the shell. The bottom of the centrifuge tube 7 is inserted into the medium solution of the convex structure; the centrifuge tube 7 is placed between two piezoelectric ceramic sheets 1. A centrifuge tube clamping groove frame 11 is arranged above the centrifuge tube clamping groove 4, and the centrifuge tube clamping groove frame 11 cooperates with the lifting clamping groove 12d of the lifting rod 12a. The centrifuge tube clamping groove is used to connect the lifting device and the centrifuge tube, and its size can be changed according to the size of the centrifuge tube. The centrifuge tube can be selected from centrifuge tubes with a capacity of 1.5mL or 2mL, but is not limited to these two types, and the size of the device shell can be changed according to the outer diameter of the centrifuge tube.

[0063] The shape of the piezoelectric ceramic piece 1 is not limited and can be circular, rectangular, elliptical, etc.; as long as the centrifuge tube 7 is in the area covered by the body acoustic wave vibration, it is required that the diameter of the centrifuge tube 7 is smaller than the distance of the piezoelectric ceramic piece 1 in the length direction of the shell side plate 2, and the bottom end of the centrifuge tube 7 is higher than the bottom end of the piezoelectric ceramic piece 1.

[0064] The top of the piezoelectric ceramic sheet 1 is tangent to or in contact with the bottom surface of the top plate 8 of the housing, so as to satisfy the distance of the biological particles inside the centrifuge tube controlled by the bulk acoustic wave to the maximum extent.

[0065] The medium solution is water, or a glycerol aqueous solution with a glycerol volume fraction of 10%-30%. When choosing a glycerol aqueous solution, the glycerol concentration range can be between 10%-30%. If the concentration is too low, the heat transfer will be faster, affecting the internal temperature of the centrifuge tube. If the concentration is too high, the viscosity will be too high, affecting the experimental efficiency.

[0066] The piezoelectric ceramic sheet 1 is provided with a piezoelectric ceramic sheet waterproof layer 1c, a piezoelectric ceramic sheet negative electrode 1a and a piezoelectric ceramic sheet positive electrode 1b in sequence from the inside to the outside of the convex structure, and the distance between the piezoelectric ceramic sheet waterproof layers (1c) on both sides is an integral multiple of the wavelength of the sound wave.

[0067] The start temperature of cooling platform 3 is 27°C and the stop temperature is 23°C.

[0068] Furthermore, the device includes: piezoelectric ceramic sheets, symmetrically placed on both sides of the device shell for generating standing waves; a convex device shell for fixing the distance between the piezoelectric ceramic sheets, with a liquid storage area and a centrifuge tube placement slot inside; a centrifuge tube slot for lifting the centrifuge tube during enrichment; a cooling platform for maintaining the temperature of the medium solution inside the device, with a temperature sensor inside the device shell to sense the internal temperature at any time to drive the cooling platform to work; a rapid lifting device for lifting the centrifuge tube after the biological particles inside the centrifuge tube are fixed; and a signal generating module for outputting a radio frequency signal to drive the piezoelectric ceramic sheets. The piezoelectric ceramic sheets are arranged on both sides of the device shell, and the distance between them is an integer multiple of the wavelength of the sound wave. The diameter of the centrifuge tube placed in the centrifuge tube slot is smaller than the diameter of the piezoelectric ceramic sheet, and the whole is in the area covered by the body sound wave vibration. The edges of the piezoelectric ceramic sheets on both sides are tangent to the internal plane of the top shell of the device. A shell for increasing the contact area with the cooling platform is arranged at the bottom of the device shell. An observation window is arranged on the side of the device shell for real-time observation of the internal enrichment situation. The internal medium solution is pure water or a glycerol aqueous solution with a concentration range of 10%-30%. The reference temperature of the cooling platform is set to 25°C, the start temperature is 27°C, and the stop temperature is 23°C.

[0069] A rapid enrichment method, comprising the following steps: injecting a medium solution into the interior of a convex device housing; placing a centrifuge tube into a centrifuge tube slot, adjusting the internal pages of the centrifuge tube to make it horizontal; adjusting a signal generation module, adjusting the input frequency and power of a radio frequency signal to start enrichment, wherein the centrifuge tube contains biological particles; lifting the centrifuge tube until the entire centrifuge tube is completely out of the working area of ​​a piezoelectric ceramic sheet. The input frequency of the radio frequency signal is 1MHz-100MHz, and the amplitude is greater than 0 and less than or equal to 100V. The lifting speed of the rapid lifting of the centrifuge tube is greater than 0 and less than or equal to 10mm / s.

[0070] The lifting speed of the centrifuge tube is 1mm / s and 0.5mm / s.

[0071] The frequency of the piezoelectric ceramic piece is 1.7 MHz, and the amplitude is 80V.

[0072] The sample liquid is human or animal body fluid or effusion, cell culture fluid, strain culture fluid or tissue culture fluid. Further, the concentration of Hela cells in the sample solution is 1-10 7 Pieces / mL.

[0073] When the above technical solution is used, the working method of the rapid enrichment device based on bulk acoustic wave includes the following steps:

[0074] In the first step, a convex structure is set on the cooling platform 3, and the piezoelectric ceramic sheets are fixedly placed on both sides of the device housing, and the positive and negative electrodes of the piezoelectric ceramic sheets on both sides are respectively connected to the signal generating module;

[0075] In the second step, the medium solution is injected into the convex structure and the liquid level exceeds the highest point of the piezoelectric ceramic sheet 1, the centrifuge tube 7 is placed in the centrifuge tube slot, and the liquid level inside the centrifuge tube is adjusted to make it horizontal; the temperature sensor is placed in the temperature sensing hole 10 on the top of the convex device shell, the medium solution temperature is monitored at any time, and the cooling platform 3 is adjusted.

[0076] The third step is to turn on the cooling platform 3 and set its temperature range to 23-27°C; when the medium solution temperature exceeds 27°C, the cooling platform starts to work and stops working after the internal temperature drops to 23°C.

[0077] The fourth step is to adjust the signal generating module, adjust the input frequency and amplitude of the RF signal; set the input frequency to 1.7 MHz, connect an oscilloscope to both ends of the piezoelectric ceramic plate 1, adjust the input voltage after amplification by the power amplifier to 80 V, and turn off the input.

[0078] Step 5: Prepare cells at a concentration of 1×10 5 1 mL of Hela cells with a concentration of 1 cell / mL is placed in a 1.5 mL centrifuge tube 7, which is then placed in the centrifuge tube slot 4. The rapid lifting device 12 is turned on and slowly lowered into the convex structure to adjust it to the level of the centrifuge tube. Figure 6 As shown in a in the figure, the lifting speed of the quick lifting device is set to 1 mm / s, the lifting mode of the quick lifting device is set to automatic lifting, and the lifting device 12 is closed.

[0079] The sixth step is to turn on the signal generating module 13 output, observe the real-time situation inside the centrifuge tube through the transparent end plate 5, and after it forms a stable line arrangement, turn on the rapid lifting device 12 again to lift it. Figure 6 As shown in b.

[0080] Step 7. Observe the lifting situation in real time during the lifting process. When the centrifuge tube leaves three quarters of the working area of ​​the piezoelectric ceramic sheet, adjust the lifting speed to 0.5 mm / s and maintain this speed until the lifting is completed.

[0081] Step 8: When the bottom of the centrifuge tube is lifted above the sonic working area, close the rapid lifting device 12 to complete the enrichment. The enrichment is completed as shown in FIG. Figure 6 As shown in middle c, the enriched Hela cells are stored at the bottom of the centrifuge tube.

[0082] Although the above are specific embodiments of the present invention, the protection scope of the present invention is not limited thereto. For those familiar with the art, any changes or substitutions made within the technical scope disclosed by the present invention should be deemed to fall within the protection scope of the present invention.

[0083] Finally, it should be noted that the protection scope of this application is not limited to the applications and technical solutions disclosed in the embodiments. Any technician familiar with the technical field can make appropriate changes and substitutions according to specific application requirements and environmental conditions within the technical scope disclosed in this application. These possible changes and substitutions, including but not limited to the selection of materials, adjustment of structures, expansion of functions, and integration with other technologies, etc., should be regarded as part of the protection scope of this application.

Claims

1. A rapid enrichment device based on bulk acoustic waves, characterized in that: The device comprises a control component, an enrichment component and a lifting device (12); the control component comprises a signal generation module (13), a temperature control module (14) and a lifting device speed adjustment knob (15); The enrichment component adopts a convex structure containing a medium solution disposed on a cooling platform (3), wherein the convex structure is a rectangular parallelepiped structure surrounded by a shell bottom plate (9), two shell side plates (2), two transparent end plates (5) and a shell top plate (8), and the shell bottom plate (9) acts on the cooling platform (3); Piezoelectric ceramic sheets (1) are respectively arranged at opposite positions of the two housing side plates (2), and the distance between the two piezoelectric ceramic sheets (1) is an integer multiple of the wavelength of the sound wave; the positive and negative electrodes of the piezoelectric ceramic sheets (1) are respectively connected to the signal generating module (13); The top plate (8) of the housing is provided with a centrifuge tube placement hole groove (6) and a temperature sensing hole (10) for installing a temperature sensor; the upper end of the centrifuge tube (7) is clamped in the centrifuge tube clamping groove (4); the centrifuge tube clamping groove (4) is placed on the top plate (8) of the housing; the bottom of the centrifuge tube (7) is inserted into the medium solution of the convex structure; the centrifuge tube (7) is placed between two piezoelectric ceramic sheets (1); a centrifuge tube clamping groove frame (11) for lifting is provided above the centrifuge tube clamping groove (4); An internal drive motor (12b) is disposed in the device base (12c) of the lifting device (12), and the internal drive motor (12b) drives the lifting rod (12a) to move up and down, and the lifting rod (12a) is provided with a lifting slot (12d) that matches the centrifuge tube slot frame (11); The cooling platform (3), the temperature sensor and the temperature control module (14) in the enrichment component are electrically connected, and the lifting device (12) is electrically connected to the lifting device speed adjustment knob (15), and the lifting device speed adjustment knob (15) is used to control the speed of the lifting rod (12a).

2. A rapid enrichment device based on bulk acoustic waves according to claim 1, characterized in that: The piezoelectric ceramic piece (1) is circular, rectangular or elliptical.

3. A rapid enrichment device based on bulk acoustic waves according to claim 2, characterized in that: The diameter of the centrifuge tube (7) is smaller than the distance of the piezoelectric ceramic sheet (1) in the length direction of the housing side plate (2), and the bottom end of the centrifuge tube (7) is higher than the bottom end of the piezoelectric ceramic sheet (1); the centrifuge tube (7) is within the area covered by the body acoustic wave vibration.

4. A rapid enrichment device based on bulk acoustic waves according to claim 3, characterized in that: The top end of the piezoelectric ceramic sheet (1) is tangent to or in contact with the bottom surface of the housing top plate (8).

5. The rapid enrichment device based on bulk acoustic wave according to claim 1, characterized in that: The medium solution is water, or a glycerol aqueous solution with a glycerol volume fraction of 10%-30%.

6. The rapid enrichment device based on bulk acoustic wave according to claim 1, characterized in that: The piezoelectric ceramic sheet (1) is provided with a piezoelectric ceramic sheet waterproof layer (1c), a piezoelectric ceramic sheet negative electrode (1a) and a piezoelectric ceramic sheet positive electrode (1b) in sequence from the inside to the outside of the convex structure, and the distance between the piezoelectric ceramic sheet waterproof layers (1c) of the two piezoelectric ceramic sheets (1) is an integer multiple of the wavelength of the sound wave.

7. The rapid enrichment device based on bulk acoustic wave according to claim 1, characterized in that: The starting temperature of the cooling platform (3) is 27°C and the stopping temperature is 23°C.

8. The working method of the rapid enrichment device based on bulk acoustic wave according to claim 1, characterized in that: The following steps are involved: S1, injecting a medium solution into the convex structure until the liquid level exceeds the highest point of the piezoelectric ceramic sheet (1); placing a temperature sensor in a temperature sensing hole (10) of the top plate (8) of the housing to monitor the temperature of the medium solution for adjusting the cooling platform (3); S2, turning on the cooling platform (3) and setting its temperature range to 23-27°C; when the temperature of the medium solution exceeds 27°C, the temperature control module (14) controls the cooling platform (3) to start working, and stops working when the temperature of the medium solution drops to 23°C; S3, adjusting the signal generating module (13) to adjust the input frequency and amplitude of the radio frequency signal; S4, loading the biological particle sample into the centrifuge tube (7), placing the centrifuge tube (7) into the centrifuge tube slot (4), placing the centrifuge tube slot rack (11) into the lifting slot (12d), lowering the centrifuge tube (7) into the convex structure, adjusting the liquid level inside the centrifuge tube (7) to make it horizontal; and setting the initial lifting speed of the lifting rod (12a) in the lifting device (12) by using the lifting device speed adjustment knob (15); S5, start the output of the signal generating module (13), observe the real-time situation inside the centrifuge tube (7) through the transparent end plate (5), and after the centrifuge tube (7) forms a stable line arrangement, start the lifting device (12); the internal driving motor (12b) drives the lifting rod (12a) to move upward, and the lifting rod (12a) further drives the centrifuge tube slot frame (11) and the centrifuge tube slot (4) to move upward, and finally drives the centrifuge tube (7) to move upward; S6. During the lifting process, the lifting situation is observed in real time. When the centrifuge tube leaves three quarters of the working area of ​​the piezoelectric ceramic sheet, the lifting speed is adjusted to half of the initial lifting speed, and the speed is maintained until the lifting is completed; S7, when the bottom of the centrifuge tube is lifted above the sonic working area, the lifting device (12) is closed to complete the enrichment. After the enrichment is completed, the biological particles are stored at the bottom of the centrifuge tube.

9. The working method of the rapid enrichment device based on bulk acoustic wave according to claim 8, characterized in that: The biological particle sample is a liquid sample containing cells, organelles or pathogens.

10. The working method of the rapid enrichment device based on bulk acoustic wave according to claim 9, characterized in that: The liquid sample is human or animal body fluid or effusion, cell culture fluid, strain culture fluid or tissue culture fluid.

Images