Acoustic flow control device for patterned arrangement of fine biological particles and use method thereof

Abstract

The invention discloses an acoustic flow control device for patterning arrangement of small biological particles and a use method thereof, the acoustic flow control device comprises a particle control chip on the upper layer and a sound source device on the lower layer, and the sound source and the particle control chip are reversibly connected through a coupling layer; a tip microstructure is arranged in the particle control chip, and the section of the microstructure is triangular; the tip microstructure adjusts sound waves excited by the ultrasonic transducer to form a vortex field converged to the tip near the microstructure, fine biological particles are enriched to the tip by means of sound stream drag force, and the patterned arrangement shape of the biological particles is regulated and controlled by controlling the position and arrangement of the tip microstructure in the particle solution storage cavity. The acoustic flow control particle patterning arrangement device is simple in structure, rapid to operate and control, small in temperature influence, capable of meeting the disposable use requirement of biological samples and low in use cost. The invention further discloses a using method of the acoustic flow control device, and efficient patterning arrangement of fine biological particles can be achieved.

Description

Technical Field

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

[0002] In recent years, the technology of manipulating biological particles such as bacteria, cells, and protozoa to arrange in patterns to achieve tissue model construction, differentiation and interaction research, drug screening and development, and other tissue engineering and biomedical applications has received widespread attention. At present, the technology of patterning biological particles based on magnetic Archimedean method, optical tweezers, self-assembly method, bioprinting and matrix templates is constantly developing, but it usually has problems such as complex equipment, cumbersome operation, low flexibility, and easy damage to particle activity, which limits its development in biochemical analysis and application fields.

[0003] Acoustofluidics combines sound waves with microfluidic chips, and uses the acoustic radiation force on particles in the fluid inside the chip to achieve particle capture, manipulation, and arrangement. Particle manipulation technology based on acoustic fluidics has the advantages of non-contact, high efficiency, low damage, simple device, and high controllability, and has great application prospects in biological sample processing. At present, acoustic fluidics technology has been rapidly developed in the manipulation of many biological particles, such as cell sorting, bacterial capture, and sorting and enrichment of extracellular vesicles. Devices based on acoustic fluidics technology mostly rely on acoustic radiation force to manipulate particles, and the shape of particle arrangement is regulated 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 control 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 2 μm), it is necessary to greatly increase the input power of the transducer to obtain the acoustic radiation force 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, which is not conducive to their application in the field of manipulation of fine biological particles, and 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 is an important development direction for acoustofluidic devices in the field of efficient manipulation and patterned arrangement of fine biological particles. Summary of the invention

[0005] In view of the above-mentioned shortcomings, the present invention provides an acoustofluidic device for patterned arrangement of fine biological particles and a method for using the same. Based on the characteristic that the drag force of the acoustic flow on the fine particles in the acoustofluidic device is greater than the acoustic radiation force, the patterned arrangement of the fine biological particles is achieved by regulating the acoustic flow field in the acoustofluidic device. The acoustofluidic patterned arrangement device of the present invention has the advantages of simple structure, little effect on the activity of biological particles, little effect by temperature, low energy consumption, short time consumption, high controllability and low cost.

[0006] In order to achieve the above-mentioned purpose, the present invention provides an acoustic fluidic 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 acoustic source device comprises an ultrasonic transducer and a coupling layer; the ultrasonic transducer is connected to the particle manipulation chip through the coupling layer; the chamber chip is sealed and connected 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 adjusts the sound waves excited by the ultrasonic transducer, and utilizes the acoustic flow effect to generate an eddy current field that converges 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 and susceptible to the heat generation and acoustic flow effects caused by high input power when applied to fine particle manipulation and patterned arrangement, and the device is not suitable for disposable use and has high cost. The present invention adopts a high-stability lead zirconate titanate piezoelectric ultrasonic transducer, and regulates the sound wave 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. The fine particles are quickly moved and converged to the tip of the microstructure in the eddy current field 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 and reduce the processing cost.

[0008] It should be noted that inside the particle solution storage chamber, eddy currents gather freely distributed small particles near the tip. When a stable state is reached, the particles present a pattern that is the same as the arrangement of the tip microstructure. By controlling the layout position and shape of the tip microstructure in the particle solution storage chamber, particle arrangements of different pattern shapes can be achieved, which has great controllability and efficiency.

[0009] The particle solution storage chamber is located on the inner surface of the chamber chip, and includes 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 avoid mixing of gas in the particle solution during injection;

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

[0011] According to one aspect of the present invention, the material of the chamber chip and the sound field modulation chip is polymethyl methacrylate or polycarbonate with good light transmittance and biocompatibility; the chamber chip and the sound field modulation chip are formed by an injection molding process and bonded and sealed by a hot pressing process; the chamber 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.

[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 is convenient for chip replacement, for patterned arrangement of different shapes and to avoid cross contamination during biological sample processing; the ultrasonic transducer is reusable to achieve 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: According to 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, and 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: Regulate the frequency and power of the ultrasonic transducer input signal to generate an eddy current field flowing toward the tip near the tip microstructure in the particle solution storage chamber, so that the freely distributed particles are quickly enriched toward the tip microstructure through the eddy current, and the patterned aggregation and arrangement of fine particles are achieved by controlling the arrangement of the tip microstructure;

[0018] Step 5: Apply a direct violet or blue light field to the chamber area where the patterned arrangement is completed to solidify the solution in the chamber, so that the patterned particles remain fixed, and remove the particle manipulation chip for subsequent observation, analysis, cultivation, etc.;

[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, and 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 transmitted to the particle manipulation chip through the coupling layer. Under the gathering and regulation of the sound wave energy by the tip microstructure, 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 acoustic radiation force and acoustic flow drag force in the acoustic and flow fields in the chamber. The magnitudes of these two forces are positively correlated with the cube and square of the particle diameter (d), respectively, 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 streaming drag on the particles is significantly greater than the acoustic radiation force. For example, when the sound pressure amplitude is 1MPa, an acoustic vortex flow field with a flow rate of 1mm / s can be generated near the microstructure by adjusting the size of the tip microstructure, and the acoustic streaming drag on particles with a diameter of 0.5μm is approximately 3 times the acoustic radiation force; under the action of a larger acoustic streaming 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 fine particles are arranged into a two-dimensional stripe pattern with a specific spacing. Taking full advantage of the fact that fine particles are greatly affected by the acoustic streaming effect, the problems of long time consumption and high energy consumption and heat generation caused by excessive input power when using only acoustic radiation force for manipulation are avoided. Adding a photocurable component to the particle solution and applying a direct purple or blue light field to the particle manipulation chip that has completed the patterned arrangement can achieve the curing 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 acoustic fluidic device of the present invention makes full use of the characteristic that small particles are greatly affected by the acoustic fluidic effect, and uses the acoustic fluidic drag force to achieve rapid manipulation and aggregation of particles at a relatively small input power. The eddy current field characteristics are regulated by controlling the size of the tip microstructure, and the shape of the particle arrangement pattern is regulated by controlling the position and arrangement shape of the tip microstructure. This 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 the acoustic field modulation chip of the acoustic fluidic device of the present invention are manufactured by injection molding and hot pressing bonding process, which has the advantages of high molding efficiency and low cost. The particle manipulation chip and the ultrasonic transducer after molding are reversibly connected. The particle manipulation chip is used as a consumable material once and the ultrasonic transducer is reused, which greatly reduces the cost of biological sample processing and contributes to the promotion and application of acoustic fluidic particle manipulation devices. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a schematic structural diagram of an acoustofluidic device for patterned arrangement of fine biological particles according to the present invention;

[0026] Figure 2 This is a schematic diagram of the structure 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 diagram of the structure 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 5A schematic cross-sectional view of the acoustic field modulation chip containing a tip microstructure according to the present invention;

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

[0031] Figure 7 It is a 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 injection port; 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 in conjunction with 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 described embodiments 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 the 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 the 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 patterned arrangement of fine biological particles is used for two-dimensional grid patterned arrangement of Neisseria gonorrhoeae (diameter 0.6-0.8 μm) in a culture medium. The schematic diagram of its structure is shown in FIG. Figure 1-Figure 4As shown. It includes an upper particle manipulation chip and a lower sound source device, wherein the particle manipulation chip includes a sound field modulation chip 3 and a cavity chip 4 made of polymethyl methacrylate (PMMA); the sound 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 sound 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 sound field modulation chip 3 is provided with a tip microstructure 32 corresponding to the particle solution storage chamber 43, and the tip microstructure 32 adjusts the sound wave excited by the ultrasonic transducer 1 to form the required sound field and flow field in the space formed by the cavity chip 4 and the sound field modulation chip 3; and the eddy current field converging to the tip is formed near the tip microstructure by using the acoustic flow effect.

[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 for discharging the gas in the chamber to avoid mixing of the gas 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 = 100μm, and the cross-section height is h = 200μm; Neisseria gonorrhoeae is arranged into a two-dimensional stripe pattern with a spacing of Δw = 80μm; the cross-section of the particle solution storage chamber is rectangular, with a height of 300μm and a width of 2mm.

[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 is convenient for chip replacement and avoids cross contamination during sample processing; the ultrasonic transducer 1 can be reused to achieve low-cost processing of samples.

[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, and controlling the injection speed of the solution so that the Neisseria gonorrhoeae sample solution fills the particle solution storage chamber to avoid residual bubbles;

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

[0045] Step 5: Apply a direct blue light field to the chamber area where the patterned arrangement is completed 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. Figure 1-Figure 4 As shown. It includes an upper particle manipulation chip and a lower sound source device, wherein the particle manipulation chip includes a sound field modulation chip 3 and a cavity chip 4 made of polycarbonate (PC); the sound 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 sound 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 sound field modulation chip 3 is provided with a tip microstructure 32 corresponding to the particle solution storage chamber 43, and the tip microstructure 32 adjusts the sound wave excited by the ultrasonic transducer 1 to form a desired sound field and flow field in the space formed by the cavity chip 4 and the sound field modulation chip 3; and an eddy current field converging to the tip is formed near the tip microstructure by using the acoustic flow effect.

[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 for discharging gas in the chamber to avoid mixing of gas 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 cross-section of the tip microstructure 32 is triangular, the bottom width of the microstructure cross-section w=80μm, and the cross-section height h=220μm; the mycoplasma is arranged in a two-dimensional stripe pattern with a spacing Δw=60μm; the cross-section of the particle solution storage chamber is rectangular, with a height of 300μm and a width of 2mm.

[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 is convenient for chip replacement and avoids cross contamination during sample processing; the ultrasonic transducer 1 can be reused to achieve low-cost processing of samples.

[0051] Method for using the above-mentioned acoustofluidic device for two-dimensional grid patterned arrangement of mycoplasma 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, and control the injection speed of the solution so that the mycoplasma sample solution fills the particle solution storage chamber to avoid residual bubbles;

[0055] Step 4: The frequency of the ultrasonic transducer input signal is adjusted to 12 MHz and the power is adjusted to 23 dBm, and an eddy current field flowing toward the tip is generated near the tip microstructure in the particle solution storage chamber, and the freely distributed mycoplasma is enriched toward the tip microstructure through the eddy current;

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

[0057] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by any technician familiar with the art within the technical scope disclosed in the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention shall be based on the protection scope of the claims.

Claims

1. An acoustofluidic device for patterned arrangement of fine biological particles, characterized in that: It comprises an upper particle manipulation chip and a lower sound source device, wherein the particle manipulation chip comprises a chamber chip and a sound 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 through the coupling layer; the chamber chip is sealed and connected to the sound 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 sound field modulation chip is provided with a tip microstructure area corresponding to the particle solution storage chamber; the tip microstructure adjusts 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; The particle solution storage chamber is located on the inner surface of the chamber chip, and includes 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 avoid mixing of gas in the particle solution during injection; Among them, the side of the acoustic field modulation chip containing a tip microstructure is bonded to the side of the chamber chip containing a 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 = 50-200μm, the cross-section height h = 100-300μm, and the spacing Δw of the tip microstructure is ≥50μm; the cross-section of the particle solution storage chamber is rectangular, the height is 200-400μm, and the width is 1-2mm, 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 chamber chip and the acoustic field modulation chip are polymethyl methacrylate or polycarbonate with good light transmittance and biocompatibility; the chamber chip and the acoustic field modulation chip are formed by an injection molding process and bonded and sealed by a hot pressing process; the chamber 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.

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, 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 is convenient for chip replacement and is used for patterned arrangements of different shapes and to avoid cross contamination during biological sample processing; the ultrasonic transducer is reusable to achieve low-cost processing of biological samples.

4. A method for using the acoustofluidic device for patterned arrangement of fine biological particles as claimed in any one of claims 1 to 3, characterized in that: The following steps are involved: Step 1: According to 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; 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 pump or a syringe, and 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: Regulate the frequency and power of the ultrasonic transducer input signal to generate an eddy current field flowing toward the tip near the tip microstructure in the particle solution storage chamber, so that the freely distributed particles are enriched toward the tip microstructure through the eddy current, and the patterned aggregation and arrangement of fine particles are achieved by controlling the arrangement of the tip microstructure; Step 5: Apply a direct violet or blue light field to the chamber area where the patterned arrangement is completed to solidify the solution in the chamber so that the patterned particles remain fixed, and remove the particle manipulation chip for subsequent observation, analysis, cultivation and other applications.

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