An acoustic fluidic device for the patterned arrangement of small biological particles and methods of use thereof

Abstract

The application discloses an acoustic flow control device for patterned arrangement of fine biological particles and a use method thereof. The acoustic flow control device comprises a particle manipulation chip in an upper layer and a sound source device in a lower layer. The sound source is reversibly connected with the particle manipulation chip through a coupling layer. The particle manipulation chip has a tip microstructure. The microstructure cross section is triangular. The tip microstructure adjusts the vortex field formed by the sound wave excited by the ultrasonic transducer to converge to the tip near the microstructure. The fine biological particles are enriched to the tip by using acoustic flow drag. The shape of the patterned arrangement of the biological particles is regulated by controlling the position and arrangement of the tip microstructure in a particle solution storage chamber. The acoustic flow control particle patterned arrangement device has the advantages of simple structure, rapid manipulation, small temperature influence, ability to meet the one-time use demand of biological samples and low use cost. The application further discloses a use method of the acoustic flow control device, and high-efficiency patterned arrangement of fine biological particles can be realized.

Description

Technical Field

[0001] The present invention relates to the field of microfluidics and biochemical particle processing technology, and in particular to an acoustofluidic device for patterned arrangement of fine biological particles and a method for using the same. Background Art

[0002] In recent years, techniques for manipulating biological particles, such as bacteria, cells, and protozoa, into patterned arrangements for tissue model construction, differentiation and interaction studies, and drug screening and development have garnered widespread attention. While techniques for patterning biological particles based on magnetic Archimedean methods, optical tweezers, self-assembly, bioprinting, and matrix templates continue to advance, they often suffer from complex equipment, cumbersome operations, limited flexibility, and potential damage to particle activity, limiting their application in biochemical analysis and applications.

[0003] Acoustofluidics, which combines sound waves with microfluidic chips, uses the acoustic radiation force exerted on particles in the fluid within the chip to achieve the capture, manipulation, and arrangement of particles. Particle manipulation technology based on acoustic fluidics has the advantages of being non-contact, highly efficient, minimally damaging, simple to install, and highly controllable, and has great application prospects in biological sample processing. Currently, acoustic fluidics has developed rapidly in the manipulation of many biological particles, such as cell sorting, bacterial capture, and the sorting and enrichment of extracellular vesicles. Devices based on acoustic fluidics mostly rely on acoustic radiation force to manipulate particles, and the shape of particle arrangement is controlled by modulating standing wave sound fields of different shapes.

[0004] At present, patterned arrangement based on acoustic radiation force is mostly applied to particles of a few microns or even tens of microns, with good manipulation effects. As the particle size decreases, the acoustic radiation force on small particles in the same sound field decreases sharply. If traditional acoustofluidic devices are used to manipulate smaller particles (such as cells and bacteria with a diameter of less than 2μm), it is necessary to greatly increase the input power of the transducer to obtain the acoustic radiation force required for manipulation, which is time-consuming and energy-consuming. At the same time, high-power input will cause serious chip heating and acoustic streaming effects, which seriously affect the arrangement and activity of biological particles. In addition, current acoustofluidic devices mostly use piezoelectric single crystals as sound sources, which have poor thermal stability and are not conducive to their application in the field of manipulation of small biological particles. In addition, the piezoelectric single crystals and chips are mostly irreversibly connected, which greatly increases the cost of biological sample processing. Therefore, how to effectively regulate and utilize the acoustic flow in acoustofluidic devices and optimize the chip structure to achieve low-cost processing are important development directions for acoustofluidic devices in the field of efficient manipulation and patterned arrangement of small biological particles. Summary of the Invention

[0005] In response to the aforementioned shortcomings, the present invention provides an acoustofluidic device for patterned arrangement of fine biological particles and a method for its use. Based on the characteristic that the drag force exerted by small particles in the acoustofluidic device is greater than the acoustic radiation force, patterned arrangement of fine biological particles is achieved by regulating the acoustic flow field within the acoustofluidic device. The acoustofluidic patterned arrangement device of the present invention has the advantages of a simple structure, minimal impact on biological particle activity, minimal temperature influence, low energy consumption, short processing time, high controllability, and low cost.

[0006] To achieve the above objectives, the present invention provides an acoustofluidic device for patterned arrangement of fine biological particles, comprising an upper particle manipulation chip and a lower sound source device, wherein the particle manipulation chip comprises a chamber chip and an acoustic field modulation chip, and the sound source device comprises an ultrasonic transducer and a coupling layer; the ultrasonic transducer is connected to the particle manipulation chip via the coupling layer; the chamber chip is sealed to the acoustic field modulation chip to form a space for particle manipulation and patterned arrangement; the chamber chip is provided with a particle solution storage chamber, an injection channel, and a waste liquid channel; the acoustic field modulation chip is provided with a tip microstructure region corresponding to the particle solution storage chamber; the tip microstructure regulates the sound waves excited by the ultrasonic transducer, and utilizes the acoustic flow effect to generate an eddy current field converging to the tip near the tip microstructure in the particle solution storage chamber;

[0007] It should be noted that the existing acoustic fluidic devices based on acoustic radiation force are time-consuming to apply to the manipulation and patterning of small particles, are susceptible to the heat generation and acoustic flow effects caused by high input power, are not suitable for disposable use, and are costly. The present invention adopts a highly stable lead zirconate titanate piezoelectric ultrasonic transducer to control the sound waves through the tip microstructure inside the particle manipulation chip, and obtains an eddy current field that converges at the tip near the tip microstructure of the particle solution storage chamber. In the eddy current field, the small particles are rapidly moved and converged to the tip of the microstructure under the action of the acoustic flow drag force. The particle manipulation chip is reversibly connected to the ultrasonic transducer through a coupling layer, thereby realizing the reuse of the ultrasonic transducer and the disposable use of the particle manipulation chip, which can avoid cross contamination during the biological sample processing process and reduce processing costs.

[0008] It should be noted that inside the particle solution storage chamber, eddy currents gather freely distributed small particles near the tip. When reaching a stable state, the particles present a pattern identical to the arrangement of the tip microstructure. By controlling the arrangement position and shape of the tip microstructure in the particle solution storage chamber, particle arrangements of different pattern shapes can be achieved, which is highly controllable and efficient.

[0009] The particle solution storage chamber is located on the inner surface of the chamber chip, including an injection inlet, an injection channel, a waste liquid channel, and a waste liquid outlet; the injection inlet is connected to the injection channel, and the waste liquid outlet is connected to the waste liquid channel; the injection channel and the waste liquid channel are respectively connected to the two ends of the particle solution storage chamber; the chamber chip is also penetrated by an injection port and a waste liquid collection port; the injection port and the waste liquid collection port are respectively connected to the injection inlet and the waste liquid outlet; the injection port and the waste liquid collection port are respectively connected to an external injection device and a collection device; the waste liquid channel and the waste liquid outlet are also used for discharging gas in the chamber to prevent gas from mixing in the particle solution during injection;

[0010] The side of the acoustic field modulation chip containing the tip microstructure is bonded to the side of the chamber chip containing the particle solution storage chamber; the tip microstructure is distributed in the area where the particle solution storage chamber is located; the cross-section of the tip microstructure is triangular, the bottom width of the microstructure cross-section w = 50-200 μm, the cross-section height h = 100-300 μm, and the spacing Δw of the tip microstructures ≥ 50 μm; the cross-section of the particle solution storage chamber is rectangular, the height is 200-400 μm, and the width is 1-2 mm, depending on the specific size required for arrangement and forming.

[0011] According to one aspect of the present invention, the material of the cavity chip and the acoustic field modulation chip is polymethyl methacrylate or polycarbonate with good light transmittance and biocompatibility; the cavity chip and the acoustic field modulation chip are formed by an injection molding process and bonded and sealed by a hot pressing process; the cavity chip and the acoustic field modulation chip have the same overall size and are positioned by boundary alignment to ensure that the tip microstructure is located inside the particle solution storage chamber after the chip is bonded.

[0012] According to one aspect of the present invention, the coupling layer is an acoustic coupling gel with a thickness of 100-200 μm; the particle manipulation chip has good overall light transmittance, a photocurable component is added to the particle solution, and an external purple or blue light source is provided to solidify and preserve the particle pattern; the connection between the particle manipulation chip and the sound source device is detachable, which facilitates chip replacement, is used for patterned arrangements of different shapes, and avoids cross-contamination during biological sample processing; the ultrasonic transducer is reusable, achieving low-cost processing of biological samples.

[0013] Based on the same inventive concept, the present invention also provides a method for using the above-mentioned acoustofluidic device for patterned arrangement of fine biological particles, comprising the following steps:

[0014] Step 1: Based on the properties and requirements of the particles to be patterned and the initial sample, select the appropriate photocurable component, appropriately dilute the initial sample, and prepare the particle solution;

[0015] Step 2: Clean the channels and chambers of the particle manipulation chip with anhydrous ethanol, deionized water, or phosphate buffer for 1-2 minutes to remove debris;

[0016] Step 3: injecting the particle solution prepared in step 1 from the injection port using a syringe pump or a syringe, controlling the injection speed of the particle solution so that the particle solution fills the particle solution storage chamber and avoids residual bubbles;

[0017] Step 4: Regulating the frequency and power of the ultrasonic transducer input signal generates an eddy current field flowing toward the tip near the tip microstructure within the particle solution storage chamber. The eddy current causes the freely distributed particles to quickly concentrate toward the tip microstructure, and by controlling the arrangement of the tip microstructure, patterned aggregation and arrangement of fine particles is achieved.

[0018] Step 5: Apply a direct violet or blue light field to the patterned chamber area to solidify the solution in the chamber, so that the patterned particles remain fixed. The particle manipulation chip is removed for subsequent observation, analysis, culture, and other applications.

[0019] The principle of patterned arrangement of tiny biological particles of the present invention:

[0020] The particle solution is injected into the injection port through a polyethylene hose using a syringe pump. The injection flow rate is controlled to fill the particle solution storage chamber with the particle solution to avoid residual bubbles. The ultrasonic transducer is used to generate plane sound waves, which are then transmitted to the particle manipulation chip through the coupling layer. Under the action of the tip microstructure on the collection and regulation of the sound wave energy, an acoustic vortex is generated near the tip microstructure and converges to the tip, such as Figure 6 As shown. Small biological particles are affected by the acoustic radiation force and acoustic drag force in the acoustic and flow fields in the chamber. The magnitude of these two forces is positively correlated with the cube and square of the particle diameter (d), that is, F 辐射 =k1d 3 , F 曳 =k2d 2 . Among them, k1 is mainly affected by the intensity of the sound field, and k2 is affected by both the intensity of the sound field and the intensity of the flow field. For small biological particles, the tip microstructure gathers the sound energy to generate a flow field in the chamber, so that when the sound field intensity is small, the acoustic flow drag on the particles is significantly greater than the acoustic radiation force. For example, when the sound pressure amplitude is 1MPa, by regulating the size of the tip microstructure, an acoustic vortex flow field with a flow rate of 1mm / s can be generated near the microstructure, and the acoustic flow drag on particles with a diameter of 0.5μm is about 3 times the acoustic radiation force; under the action of a larger acoustic flow drag, the small particles move rapidly with the flow field, and at this time the particle trajectory is mainly determined by the flow field state. Regulating the sound field input and microstructure parameters can gather small particles near any tip microstructure in the chamber to the tip. Therefore, by controlling the position and arrangement shape of the tip microstructure, the small particles in the chamber can be patterned according to a specific shape, such as Figure 7 As shown, the tiny particles are arranged into a two-dimensional stripe pattern with a specific spacing. By fully utilizing the fact that tiny particles are greatly affected by the acoustic streaming effect, the long manipulation time and high energy consumption and heat generation caused by excessive input power when using acoustic radiation force alone are avoided. Adding a photocurable component to the particle solution and applying a direct violet or blue light field to the particle manipulation chip that has completed the patterned arrangement can achieve the solidification and preservation of the patterned arrangement.

[0021] Beneficial effects of the present invention:

[0022] (1) The acoustofluidic device for patterned arrangement of fine biological particles of the present invention has a simple structure, easy operation, and high stability. It also meets the requirements of disposable use and low cost for biological sample processing and has broad application prospects in the field of patterned arrangement of biological particles.

[0023] (2) The acoustofluidic device of the present invention makes full use of the characteristic that small particles are greatly affected by the acoustofluidic effect, and uses the acoustofluidic drag force to achieve rapid manipulation and aggregation of particles at a relatively low input power. By controlling the size of the tip microstructure to regulate the eddy current field characteristics, and controlling the position and arrangement shape of the tip microstructure to regulate the shape of the arrangement pattern of the particles, it greatly avoids the adverse effects of heat generation and additional flow fields caused by long time and high power consumption, and has good biological application value.

[0024] (3) The chamber chip and acoustic field modulation chip of the acoustofluidic device of the present invention are manufactured through injection molding and hot-press bonding processes, which have the advantages of high molding efficiency and low cost. The particle manipulation chip and ultrasonic transducer are reversibly connected after molding. The particle manipulation chip is disposable as a consumable material, while the ultrasonic transducer is reusable. This greatly reduces the cost of biological sample processing and contributes to the promotion and application of acoustofluidic particle manipulation devices. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Schematic diagram of the structure of an acoustofluidic device for patterned arrangement of fine biological particles according to the present invention;

[0026] Figure 2 This is a schematic structural diagram of the inner surface of the chamber chip of the acoustofluidic device for patterned arrangement of fine biological particles according to the present invention;

[0027] Figure 3 This is a schematic structural diagram of the outer surface of the chamber chip of the acoustofluidic device for patterned arrangement of fine biological particles according to the present invention;

[0028] Figure 4 This is a schematic structural diagram of the inner surface of the acoustic field modulation chip of the acoustofluidic device for patterned arrangement of fine biological particles according to the present invention;

[0029] Figure 5Schematic cross-sectional view of the acoustic field modulation chip containing the cutting-edge microstructure according to the present invention;

[0030] Figure 6 Schematic diagram of the force and movement trend of fine particles in the particle solution storage chamber containing the tip microstructure of the present invention;

[0031] Figure 7 Schematic diagram of the effect of arrangement of fine particles in the particle solution of the present invention.

[0032] Description of reference numerals:

[0033] 1. Ultrasonic transducer; 2. Coupling layer; 3. Acoustic field modulation chip; 4. Chamber chip; 31. Inner surface of acoustic field modulation chip; 32. Tip microstructure; 41. Sample inlet; 42. Sample injection channel; 43. Particle solution storage chamber; 44. Waste liquid outlet; 45. Waste liquid channel; 46. Inner surface of chamber chip; 47. Sample inlet; 48. Waste liquid collection port; 49. Outer surface of chamber chip. DETAILED DESCRIPTION

[0034] To make the present invention easier to understand, the present invention is further described below with reference to specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. Unless otherwise defined, the professional terms used below are consistent with the meanings understood by professional and technical personnel in this field; unless otherwise specified, the raw materials and reagents involved in this article can be purchased from the market or prepared by known methods.

[0035] Example 1

[0036] An acoustic fluidic device for patterning small biological particles is used for two-dimensional grid patterning of Neisseria gonorrhoeae (diameter 0.6-0.8 μm) in culture medium. The schematic diagram of its structure is shown in FIG. Figures 1-4As shown. It includes an upper particle manipulation chip and a lower sound source device. The particle manipulation chip includes an acoustic field modulation chip 3 and a cavity chip 4 made of polymethyl methacrylate (PMMA). The acoustic field modulation chip 3 and the cavity chip 4 are manufactured by injection molding and bonded by a hot pressing process. The sound source device includes an ultrasonic transducer 1 and a coupling layer 2. The ultrasonic transducer 1 and the acoustic field modulation chip 3 are reversibly connected via the coupling layer 2. The ultrasonic transducer 1 is a lead zirconate titanate piezoelectric ceramic (PZT) with an impedance matching layer. The cavity chip 4 is provided with a particle solution storage chamber 43. The acoustic field modulation chip 3 is provided with a tip microstructure 32 corresponding to the particle solution storage chamber 43. The tip microstructure 32 regulates the sound waves excited by the ultrasonic transducer 1 to form the desired acoustic field and flow field within the space formed by the cavity chip 4 and the acoustic field modulation chip 3. The acoustic flow effect is used to form an eddy current field near the tip microstructure that converges to the tip.

[0037] The particle solution storage chamber 43 is located on the inner surface 46 of the chamber chip, and includes an injection inlet 41, an injection channel 42, a waste liquid channel 45, and a waste liquid outlet 44; the injection inlet 41 is connected to the injection channel 42, and the waste liquid outlet 44 is connected to the waste liquid channel 45; the injection channel 42 and the waste liquid channel 45 are respectively connected to the two ends of the particle solution storage chamber 43; the chamber chip 4 is also penetrated by an injection port 47 and a waste liquid collection port 48; the injection port 47 and the waste liquid collection port 48 are respectively connected to the injection inlet 41 and the waste liquid outlet 44; the injection port 47 and the waste liquid collection port 48 are respectively connected to the external injection device and the collection device; the waste liquid channel 45 and the waste liquid outlet 44 are also used to discharge the gas in the chamber to prevent the gas from mixing in the Neisseria gonorrhoeae sample solution during injection.

[0038] The side of the acoustic field modulation chip 3 containing the tip microstructure 32 is bonded to the side of the chamber chip 4 containing the particle solution storage chamber 43; the tip microstructure 32 is distributed in the area where the particle solution storage chamber 43 is located; the cross-section of the tip microstructure 32 is triangular, the bottom width of the microstructure cross-section is w = 100 μm, and the cross-section height is h = 200 μm; Neisseria gonorrhoeae is arranged into a two-dimensional stripe pattern with a spacing of Δw = 80 μm; the cross-section of the particle solution storage chamber is rectangular, with a height of 300 μm and a width of 2 mm.

[0039] The coupling layer 2 is an acoustic coupling gel with a thickness of 150 μm. The particle manipulation chip has good overall light transmittance. Diluted methacrylated gelatin and a matching photoinitiator are added to the particle solution, and an external direct blue light source is provided to solidify and preserve the arrangement pattern of Neisseria gonorrhoeae. The connection between the particle manipulation chip and the ultrasonic transducer 1 is detachable, which facilitates chip replacement and avoids cross contamination during sample processing. The ultrasonic transducer 1 is reusable, achieving low-cost sample processing.

[0040] Method for using the above-mentioned acoustofluidic device for two-dimensional grid patterned arrangement of Neisseria gonorrhoeae in culture medium:

[0041] Step 1: diluting a Neisseria gonorrhoeae sample containing a photocurable component with a phosphate buffer solution to prepare a Neisseria gonorrhoeae sample solution for patterned arrangement;

[0042] Step 2: Clean the channels and chambers of the particle manipulation chip with anhydrous ethanol and phosphate buffer for 2 minutes to remove debris;

[0043] Step 3: injecting the Neisseria gonorrhoeae sample solution prepared in step 1 from the injection port using a syringe pump, controlling the injection speed of the solution so that the Neisseria gonorrhoeae sample solution fills the particle solution storage chamber and avoids residual bubbles;

[0044] Step 4: The frequency of the ultrasonic transducer input signal is adjusted to 10 MHz and the power to 20 dBm, generating an eddy current field flowing toward the tip near the tip microstructure in the particle solution storage chamber, and the eddy current causes the freely distributed Neisseria gonorrhoeae to be enriched toward the tip microstructure;

[0045] Step 5: Apply a direct blue light field to the patterned chamber area to solidify the solution in the chamber, so that the patterned Neisseria gonorrhoeae remains fixed, and remove the particle manipulation chip for subsequent observation, analysis, culture, and other applications.

[0046] Example 2

[0047] An acoustic fluidic device for patterned arrangement of fine biological particles is used for patterned arrangement of mycoplasma (diameter 0.1-0.3 μm) in culture medium. The schematic diagram of its structure is shown in FIG. Figures 1-4 As shown. It includes an upper particle manipulation chip and a lower sound source device. The particle manipulation chip includes an acoustic field modulation chip 3 and a cavity chip 4 made of polycarbonate (PC). The acoustic field modulation chip 3 and the cavity chip 4 are manufactured by injection molding and bonded by a hot pressing process. The sound source device includes an ultrasonic transducer 1 and a coupling layer 2. The ultrasonic transducer 1 and the acoustic field modulation chip 3 are reversibly connected through the coupling layer 2. The ultrasonic transducer 1 is a lead zirconate titanate piezoelectric ceramic (PZT) with an impedance matching layer. The cavity chip 4 is provided with a particle solution storage chamber 43, and the acoustic field modulation chip 3 is provided with a tip microstructure 32 corresponding to the particle solution storage chamber 43. The tip microstructure 32 regulates the sound waves excited by the ultrasonic transducer 1 to form the desired sound field and flow field in the space formed by the cavity chip 4 and the acoustic field modulation chip 3. The acoustic streaming effect is used to form an eddy current field near the tip microstructure that converges to the tip.

[0048] The particle solution storage chamber 43 is located on the inner surface 46 of the chamber chip, and includes an injection inlet 41, an injection channel 42, a waste liquid channel 45, and a waste liquid outlet 44; the injection inlet 41 is connected to the injection channel 42, and the waste liquid outlet 44 is connected to the waste liquid channel 45; the injection channel 42 and the waste liquid channel 45 are respectively connected to the two ends of the particle solution storage chamber 43; the chamber chip 4 is also penetrated by an injection port 47 and a waste liquid collection port 48; the injection port 47 and the waste liquid collection port 48 are respectively connected to the injection inlet 41 and the waste liquid outlet 44; the injection port 47 and the waste liquid collection port 48 are respectively connected to an external injection device and a collection device; the waste liquid channel 45 and the waste liquid outlet 44 are also used to discharge the gas in the chamber to prevent the gas from mixing in the mycoplasma sample solution during injection.

[0049] The side of the acoustic field modulation chip 3 containing the tip microstructure 32 is bonded to the side of the chamber chip 4 containing the particle solution storage chamber 43; the tip microstructure 32 is distributed in the area where the particle solution storage chamber 43 is located; the tip microstructure 32 has a triangular cross-section, with a base width w = 80 μm and a cross-section height h = 220 μm; the mycoplasmas are arranged in a two-dimensional stripe pattern with a spacing Δw = 60 μm; the particle solution storage chamber has a rectangular cross-section with a height of 300 μm and a width of 2 mm.

[0050] The coupling layer 2 is an acoustic coupling gel with a thickness of 150 μm. The particle manipulation chip has good overall light transmittance. Diluted methacrylated gelatin and a matching photoinitiator are added to the particle solution, and an external direct blue light source is provided to solidify and preserve the mycoplasma arrangement pattern. The connection between the particle manipulation chip and the ultrasonic transducer 1 is detachable, which facilitates chip replacement and avoids cross contamination during sample processing. The ultrasonic transducer 1 is reusable, achieving low-cost sample processing.

[0051] Method for using the above-mentioned acoustofluidic device for two-dimensional grid patterned arrangement of mycoplasmas in culture medium:

[0052] Step 1: diluting a mycoplasma sample containing a photocurable component with a phosphate buffer solution to prepare a mycoplasma sample solution for patterned arrangement;

[0053] Step 2: Clean the channels and chambers of the particle manipulation chip with anhydrous ethanol and phosphate buffer for 2 minutes to remove debris;

[0054] Step 3: Use a syringe pump to inject the mycoplasma sample solution prepared in step 1 from the injection port. Control the injection speed of the solution so that the mycoplasma sample solution fills the particle solution storage chamber and avoids residual bubbles.

[0055] Step 4: The frequency of the ultrasonic transducer input signal was adjusted to 12 MHz and the power to 23 dBm. This generated an eddy current field flowing toward the tip near the tip microstructure within the particle solution storage chamber. The eddy current enriched the freely distributed mycoplasmas toward the tip microstructure.

[0056] Step 5: Apply a direct blue light field to the patterned chamber area to solidify the solution in the chamber, so that the patterned mycoplasma remains fixed. Remove the particle manipulation chip for subsequent observation, analysis, culture, and other applications.

[0057] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be easily conceived by a person skilled in the art within the technical scope disclosed in the present invention should be included in the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be based on the scope of protection of the claims.

Claims

1. An acoustofluidic device for patterning small biological particles, characterized in that: The invention comprises an upper particle manipulation chip and a lower sound source device, wherein the particle manipulation chip comprises a chamber chip and an acoustic field modulation chip, and the acoustic source device comprises an ultrasonic transducer and a coupling layer; the ultrasonic transducer is connected to the particle manipulation chip via the coupling layer; the chamber chip is sealed to the acoustic field modulation chip to form a space for particle manipulation and patterned arrangement; the chamber chip is provided with a particle solution storage chamber, an injection channel, and a waste liquid channel; the acoustic field modulation chip is provided with a tip microstructure area corresponding to the particle solution storage chamber; the tip microstructure regulates the sound waves excited by the ultrasonic transducer, and utilizes the acoustic streaming effect to generate an eddy current field converging to the tip near the tip microstructure in the particle solution storage chamber; The particle solution storage chamber is located on the inner surface of the chamber chip, including an injection inlet, an injection channel, a waste liquid channel, and a waste liquid outlet; the injection inlet is connected to the injection channel, and the waste liquid outlet is connected to the waste liquid channel; the injection channel and the waste liquid channel are respectively connected to the two ends of the particle solution storage chamber; the chamber chip is also penetrated by an injection port and a waste liquid collection port; the injection port and the waste liquid collection port are respectively connected to the injection inlet and the waste liquid outlet; the injection port and the waste liquid collection port are respectively connected to an external injection device and a collection device; the waste liquid channel and the waste liquid outlet are also used for discharging gas in the chamber to prevent gas from mixing in the particle solution during injection; Among them, the side of the acoustic field modulation chip containing the tip microstructure is bonded to the side of the chamber chip containing the particle solution storage chamber; the tip microstructure is distributed in the area where the particle solution storage chamber is located; the cross-section of the tip microstructure is triangular, the bottom width of the microstructure cross-section w=50-200 μm, the cross-section height h=100-300 μm, and the spacing Δw of the tip microstructure is ≥50 μm; the cross-section of the particle solution storage chamber is rectangular, the height is 200-400 μm, and the width is 1-2 mm, depending on the specific size required for arrangement and forming.

2. The acoustofluidic device for patterned arrangement of fine biological particles according to claim 1, characterized in that: The materials of the cavity chip and the sound field modulation chip are polymethyl methacrylate or polycarbonate; the cavity chip and the sound field modulation chip are formed by an injection molding process and bonded and sealed by a hot pressing process; the cavity chip and the sound field modulation chip have the same overall size and are positioned by boundary alignment to ensure that the tip microstructure is located inside the particle solution storage chamber after the chip is bonded.

3. The acoustofluidic device for patterned arrangement of fine biological particles according to claim 1, characterized in that: The coupling layer is an acoustic coupling gel with a thickness of 100-200 μm. The particle manipulation chip has good overall light transmittance. The particle solution storage chamber is used to accommodate a particle solution added with a photocurable component, and is equipped with an external purple or blue light source to irradiate the cavity area where the patterned arrangement is completed to solidify the solution in the cavity and keep the patterned particles fixed.

4. A method for using an acoustofluidic device for patterned arrangement of fine biological particles, wherein the acoustofluidic device is the acoustofluidic device for patterned arrangement of fine biological particles according to claim 1, characterized in that: The following steps are involved: Step 1: Based on the properties and requirements of the particles to be patterned and the initial sample, select a suitable photocurable component, appropriately dilute the initial sample, and prepare a particle solution containing the photocurable component; Step 2: Clean the channels and chambers of the particle manipulation chip with anhydrous ethanol, deionized water, or phosphate buffer for 1-2 minutes to remove debris; Step 3: injecting the particle solution prepared in step 1 from the injection port using a syringe, controlling the injection speed of the particle solution so that the particle solution fills the particle solution storage chamber and avoids residual bubbles; Step 4: Regulating the frequency and power of the ultrasonic transducer input signal generates an eddy current field flowing toward the tip near the tip microstructure within the particle solution storage chamber. The eddy current causes the freely distributed particles to be concentrated toward the tip microstructure, and by controlling the arrangement of the tip microstructure, patterned aggregation and arrangement of fine particles is achieved. Step 5: Apply a direct violet or blue light field to the patterned chamber area to solidify the solution in the chamber, so that the patterned particles remain fixed, and remove the particle manipulation chip for subsequent observation, analysis, and culture applications.

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