CN112973986A

CN112973986A - Centrifugal device

Info

Publication number: CN112973986A
Application number: CN201911286626.2A
Authority: CN
Inventors: 郑海荣; 孟龙; 赵泽胤; 崔笑宇; 牛丽丽
Original assignee: Shenzhen Institute of Advanced Technology of CAS
Current assignee: Shenzhen Institute of Advanced Technology of CAS
Priority date: 2019-12-14
Filing date: 2019-12-14
Publication date: 2021-06-18
Anticipated expiration: 2039-12-14
Also published as: CN112973986B

Abstract

The invention relates to the technical field of biomedical engineering, and discloses a centrifugal device, which comprises a glass slide, a signal generator, a power amplifier and a transducer, wherein the signal generator is arranged on the glass slide; the glass slide is adhered with a micropore structure and a cavity structure which are mutually bonded, and is provided with the transducer; the cavity structure is used for introducing liquid to be separated; the signal generator is used for generating an electric signal, the power amplifier is used for amplifying the electric signal, and the transducer is used for converting the amplified electric signal into an ultrasonic signal and acting on the microporous structure; and the liquid to be separated passes through the microporous structure, forms microbubbles under the action of the ultrasonic signal and generates resonance, so that the separation of different particles in the liquid to be separated is realized. The invention has simple structure, high efficiency, higher biological safety, higher result consistency, automation and high repeatability, and the structure of the product obtained by separation is complete.

Description

Centrifugal device

Technical Field

The invention relates to the technical field of biomedical engineering, in particular to a centrifugal device.

Background

Exosomes are nanoscale vesicle structures that can be actively secreted by both normal cells and tumor cells, and have a diameter of about 30-150 nm. Exosomes are widely and stably present in a variety of clinical samples, including blood, urine, ascites, interstitial fluid, tears, saliva, and cerebrospinal fluid, among others. Substances contained in exosomes from different sources are different, and the exosomes provide good biological materials for researching potential various biomarkers. Exosomes possess specific proteins of their mother cells, which also contain various nucleic acid molecules, and thus, the various specific proteins and nucleic acid molecules carried by exosomes provide abundant detectable targets for disease analysis. Exosomes participate in regulating important cell physiological activities, have reports on the effects in immune response, apoptosis, angiogenesis, inflammatory reaction and coagulation processes, can be used as early diagnosis markers of various diseases, and can also be used as carriers of targeted drugs for disease treatment. And some exosome-based tumor therapies are already in clinical trials. At present, the main obstacle to realizing clinical application of exosomes is the lack of a standard method for quickly, stably and efficiently extracting high-purity exosomes.

With the increasing maturity of micro-nano processing technology, micro-fluidic chips (also called Lab-on-a-chips) in which microbubbles are widely used, such as liquid mixing (Ahmed D, Lab-on-a-chips, 2009,9(18): 2738-. The device operates by trapping gas bubbles within a "horseshoe" structure located inside the microchannel between two laminar flows. After the single horseshoe-shaped structure in the cavity channel is immersed in liquid, an air-liquid film is formed, and the micro-bubble vibration generates a micro-flow field for realizing liquid mixing.

The separation of different particle sizes is achieved by acoustically manipulating the chip (Mengxi W, PNAS,201709210), this method consists of the integration of two sequential Surface Acoustic Wave (SAW) microfluidic modules, the platform being capable of separating exosomes directly from undiluted whole blood samples. Micro blood components, including red blood cells, white blood cells and platelets, greater than 1 μm in diameter were extracted by the cell removal module to obtain EV-rich samples. Removal of EV and apoptotic bodies was achieved by the exosome-separating module, obtaining samples with purified exosomes. Each module relies on the oblique-angle saw surface wavefield.

Existing methods for exosome purification include ultracentrifugation, size exclusion chromatography, magnetic bead-based immunoaffinity capture, polyethylene glycol-based precipitation, ultrafiltration, and microfluidics. However, the existing techniques are not ideal and limit the clinical transformation and application of exosomes. Ultracentrifugation is the most common method for purifying exosomes, but it has low yield (recovery rate: 5% -25%), complicated operation process, long time (more than 1 day), and depends on expensive equipment. Because of the ultra-high speed centrifugation, hemolysis is easy to occur, and the experimental result is influenced. The size exclusion chromatography is based on high cost, low recovery rate and low purity. Immunocapture-based separation methods can collect exosomes with medium or high purity, but are limited by the specificity of the antibody and the cumbersome procedures, difficult to standardize, and not suitable for handling large quantities and volumes of clinical samples. Polyethylene glycol precipitation-based methods are time consuming, have many impurities, require biomarkers, and have poor exosome integrity. The ultrafiltration method is easy to block and has low flux. The separation method based on the microfluidic technology cannot solve the problems of low copper beam, complex operation process and poor repeatability, and is difficult to realize the structural consistency among laboratories.

Disclosure of Invention

The invention aims to solve the technical problems in the prior art and provide a centrifugal device which is simple and efficient in structure, higher in biosafety and result consistency, automatic and high in repeatability, and the obtained product is completely separated.

In order to solve the problems proposed above, the technical scheme adopted by the invention is as follows:

a centrifuge device comprising a slide, a signal generator, a power amplifier and a transducer; the glass slide is adhered with a micropore structure and a cavity structure which are mutually bonded, and is provided with the transducer;

the cavity structure is used for introducing liquid to be separated; the signal generator is used for generating an electric signal, the power amplifier is used for amplifying the electric signal, and the transducer is used for converting the amplified electric signal into an ultrasonic signal and acting on the microporous structure; and the liquid to be separated passes through the microporous structure, forms microbubbles under the action of the ultrasonic signal and generates resonance, so that the separation of different particles in the liquid to be separated is realized.

Further, the microporous structure is seamlessly attached to the glass slide, the cavity structure is arranged on the microporous structure, and the cavity structure and the microporous structure are bonded; and the corresponding positions of the glass slide, the micropore structure and the cavity structure are respectively provided with a positioning structure.

Furthermore, the micropore structure comprises a uniform-diameter array of micro-bubble structures, and the micro-bubbles in two adjacent rows are staggered.

Furthermore, the cavity structure comprises microfluidic channels with symmetrical structures, all the microfluidic channels are distributed in a snake shape, and two ends of each microfluidic channel are respectively used as a sample inlet end and a sample outlet end.

Further, the array range of the micro-bubbles is larger than the distribution range of the micro-fluidic cavities.

Furthermore, the micropore structure and the cavity structure are both made of PDMS chips by soft lithography and mold copying technology.

Further, the manufacturing process of the micropore structure and the cavity structure comprises the following steps:

step S1: selecting two silicon wafers, placing the two silicon wafers in pure alcohol solution for cleaning, drying the silicon wafers by using nitrogen, and placing the silicon wafers on a hot plate for baking and cooling;

step S2: placing the cleaned and dried silicon wafer on a spin coater, adding negative photoresist for spin coating, and placing the silicon wafer on a hot plate for baking and cooling;

step S3: respectively placing a film containing a microfluidic channel and a film containing a micropore array structure right above a photoresist area on a silicon chip, carrying out exposure treatment on the photoresist area through a photoetching machine, and placing the photoresist area on a hot plate for baking and cooling;

step S4: immersing the exposed silicon wafer into SU-8 developing solution, shaking a glass vessel, spraying the developing solution and isopropanol for washing, drying by using nitrogen, and placing the silicon wafer on a hot plate for baking and cooling to obtain two silicon wafers respectively containing a microfluidic cavity and a micropore array structure;

step S5: uniformly mixing a PDMS main agent and a hardening agent according to a proportion to obtain a mixture, and respectively pouring the mixture into the silicon chip containing the microfluidic cavity channel and the silicon chip containing the micropore array structure;

step S6: vacuumizing the silicon wafer to remove bubbles in PDMS, and curing;

step S7: removing the cured PDMS, and punching a hole on a silicon wafer containing the microfluidic channel by using a puncher to serve as an inlet end and an outlet end of the liquid to be detected;

step S8: and bonding two silicon wafers respectively containing a micro-fluidic cavity channel and a micropore array structure together to obtain a PDMS micro-fluidic chip, and attaching the PDMS micro-fluidic chip to the glass slide.

Further, the glass slide is a high-light-transmission medical glass slide.

Further, the transducer adopts a PZT piezoelectric transducer.

Compared with the prior art, the invention has the beneficial effects that:

the invention directly separates the liquid to be separated through the micropore structure and the cavity structure without dilution, generates an excitation signal through the signal generator, the power amplifier and the transducer to act on the microbubbles to generate resonance, performs operation and screening without direct contact and damage, can centrifuge various gradient nanometer micron-level substances in real time by adjusting different power energies, has the characteristics of simple structure, low input energy, low cost, high timeliness and the like, can also directly separate the required substances from biological fluid on a single device in an automatic mode, and has high purity of the separated substances, high yield and complete structure.

Drawings

FIG. 1 is a schematic view of the overall structure of the centrifugal apparatus of the present invention.

FIG. 2 is a plan view of a slide glass of the present invention.

FIG. 3 is a plan view showing the structure of the micro-pores according to the present invention.

FIG. 4 is a plan view of the channel structure of the present invention.

FIG. 5 is a schematic diagram of the structure of microbubbles according to the invention.

FIG. 6 is a conceptual diagram of the cell capture by the array microbubble of the present invention.

FIG. 7 is a flow chart of the fabrication of the microporous structure and the channel structure of the present invention.

FIG. 8 is a schematic view of the fabrication of the micro-porous structure and the channel structure of the present invention.

FIG. 9 shows the adsorption and release of PS beads in the present invention under the operation of the chip.

FIG. 10 is a graph showing the relationship between the particle size and the amount of the substance obtained at the outlet end of the sample according to the present invention.

The drawings illustrate the following: 10-glass slide, 20-signal generator, 30-power amplifier, 40-transducer, 50-micropore structure, 60-channel structure, 11, 12, 13, 14-first positioning structure, 51, 52, 53, 54-second positioning structure, 61, 62, 63, 64-third positioning structure, 501-microbubble structure and 601-microfluidic channel.

Detailed Description

To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Preferred embodiments of the present invention are shown in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

Referring to fig. 1, the embodiment of the present invention provides a centrifugal device, which includes a slide 10, a signal generator 20, a power amplifier 30, and a transducer 40; the micro-pore structure 50 and the cavity structure 60 are attached to the glass slide 10, and the transducer 40 is arranged on the glass slide; the microporous structure 50 and the channel structure 60 are bonded.

The cavity structure 60 is used for introducing liquid to be separated; the signal generator 20 is used for generating an electrical signal, the power amplifier 30 is used for amplifying the electrical signal, and the transducer 40 is used for converting the amplified electrical signal into an ultrasonic signal and acting on the microporous structure 50; the liquid to be separated passes through the microporous structure 50, forms microbubbles under the action of the ultrasonic signal and generates resonance, so that the separation of different particles in the liquid to be separated is realized.

In the centrifugal device provided by the embodiment of the present invention, after the liquid to be separated is introduced into the cavity structure 60 by an external low pressure peristaltic pump, due to the surface tension of the liquid, the liquid to be separated flows through the microporous structure 50, and then under the excitation of an ultrasonic signal, a liquid-air film is formed and microbubbles are formed, so that a pressure difference exists between the air surrounded by the liquid to be separated and the surrounding liquid, so that the formed microbubbles generate stable cavitation, and resonance occurs to realize separation of different particles in the liquid to be separated. Because the resonance frequency of the micro-bubble depends on the magnitude of the input energy, in the embodiment of the present invention, the capturing of different particles can be achieved by adjusting the energy of the electrical signal input by the signal generator 20 or changing the vibration amplitude of the micro-bubble by the frequency, and the separation of different nano-or micron-scale particle sizes can be achieved by adjusting different amplitudes of the signal generator 20 to change the resonance of the micro-bubble.

In the embodiment of the present invention, the microporous structure 50 is seamlessly attached to the slide 10, and the channel structure 60 is disposed on the microporous structure 50 and bonded to the microporous structure 50, so that the ultrasonic signal can reliably and effectively act on the microporous structure 50. In order to ensure that the microporous structure 50, the channel structure 60 and the slide glass 10 can be seamlessly bonded together, the corresponding positions of the microporous structure 50, the channel structure 60 and the slide glass 10 are respectively provided with a positioning structure, as shown in fig. 2 to 4, that is, the slide glass 10 is provided with a first positioning structure (11, 12, 13, 14), the microporous structure 50 is provided with a second positioning structure (51, 52, 53, 54), the channel structure 60 is provided with a third positioning structure (61, 62, 63, 64), and the structure is simple and can realize quick and accurate bonding.

In the embodiment of the present invention, the microporous structure 50 includes uniform-diameter array-distributed micro-bubble structures 501, and two adjacent rows of the micro-bubble structures 501 are staggered (see fig. 5), so that substances in a liquid can be uniformly distributed in the micro-fluidic channel, and large substances in the liquid can be sufficiently and uniformly captured by bubbles, thereby reliably realizing large-scale particle screening. Further, the diameter of the micro-bubble structure 501 is preferably 40 μm, and since the micro-bubble vibration amplitudes of different diameters are different, the disturbance to the surrounding environment is different, and the velocities of particles with different diameters in the same micro-bubble field are different, the particles with different diameters can be screened by changing the diameter of the micro-bubble structure 501.

In the embodiment of the present invention, the channel structure 60 includes microfluidic channels 601 with symmetrical structures, all the microfluidic channels 601 are distributed in a serpentine manner, and two ends of each microfluidic channel 601 are respectively used as a sample inlet end 2 and a sample outlet end 8. The microfluidic channel 601 adopts a symmetrical structure and is distributed in a snake shape, so that liquid can be fully filtered to remove larger substances in the microfluidic channel 601 when the liquid is introduced, and the separation reliability is ensured. The microfluidic channel 601 can contain blood, urine or other liquid to be separated.

Furthermore, two ends of the microfluidic channel 601 are perforated by a puncher to serve as a sample inlet end 2 and a sample outlet end 8, and the liquid to be separated is injected into the microfluidic channel 601 through the sample inlet end 1 by a syringe through a hose, so that the safety and reliability of the whole separation process can be ensured, and the whole separation process is not polluted.

In the embodiment of the present invention, the array range of the micro-bubbles 501 on the microporous structure 50 is larger than the distribution range of the microfluidic channel 601, so that the two are reliably bonded.

In the embodiment of the invention, the glass slide 10 is a high-light-transmission medical glass slide with good effect.

In the embodiment of the present invention, the transducer 40 employs a PZT piezoelectric transducer, which can reliably and effectively convert an electrical signal into an ultrasonic signal.

Fig. 6 is a conceptual diagram of capturing cells by the array microbubble of the present invention, wherein the bubbles are microbubbles, and the cells are particles in the liquid to be separated, specifically:

after the liquid to be separated is injected into the microfluidic channel 601, the liquid to be separated forms stable microbubbles when passing through the microbubble structure 501 of the array, and when the microbubbles are excited by a sound field with a wavelength much larger than the diameter of the microbubbles, the microbubbles vibrate to generate microflows. The surrounding fluid medium is influenced through the respiration effect of the surface membrane, and the energy is transferred to particles (namely, substances to be separated) in the microfluidic channel 601, so that the particles are controlled. Under the vibration action of the micro-bubbles, the particles in the micro-fluidic cavity 601 are mainly acted by acoustic radiation force and micro-flow induced drag force, and the incident waves generated by the resonance micro-bubbles are scattered to cause the particles to be acted by the radiation force, namely the force generated by the micro-bubbles can adsorb the particles in the liquid to be separated. The distance between the particles and the microbubbles in the microfluidic channel 601 greatly affects the size of the acoustic radiation force, and theoretically, when the density of the particles is greater than that of the liquid, the particles are attracted. When the density of the particles in the microfluidic channel 601 is less than that of the liquid, the particles are repelled. When the liquid density and the density of the particles are equal, the radiation force to which the particles are subjected is very small. The microfluidic radiation force increases with the increase of the diameter of the microbubble, and the particle diameter is positively correlated with the radiation force.

In the embodiment of the present invention, the microporous structure 50 and the channel structure 60 are both made of PDMS (polydimethylsiloxane) chips by using soft lithography and mold copying techniques, and as shown in fig. 7, the specific manufacturing process includes the following steps:

step S1: selecting two silicon wafers, placing the two silicon wafers in pure alcohol solution for cleaning, drying the two silicon wafers by using nitrogen, and placing the two silicon wafers on a hot plate for baking and cooling.

Step S2: and (3) placing the cleaned and dried silicon wafer on a spin coater, adding negative photoresist for spin coating, and placing the silicon wafer on a hot plate for baking and cooling.

Step S3: respectively placing a film containing a microfluidic channel and a film containing a micropore array structure right above a photoresist area on a silicon chip, carrying out exposure treatment on the photoresist area through a photoetching machine, and placing the photoresist area on a hot plate for baking and cooling.

Step S4: immersing the exposed silicon wafer into SU-8 developing solution, shaking the glass vessel, spraying and washing the residual photoresist by using the developing solution, washing and removing the residual developing solution by using isopropanol, drying by using nitrogen, and placing the silicon wafer on a hot plate for baking and cooling to obtain two silicon wafers respectively containing a microfluidic cavity channel and a micropore array structure.

Step S5: and uniformly mixing the PDMS main agent and the hardening agent in proportion to obtain a mixture, and pouring the mixture into the silicon chip containing the microfluidic cavity and the silicon chip containing the micropore array structure respectively.

Step S6: and vacuumizing the silicon wafer to remove air bubbles in the PDMS, and curing.

Step S7: and (3) removing the cured PDMS, and punching holes on a silicon wafer containing the microfluidic channel by using a puncher to serve as an inlet end and an outlet end of the liquid to be detected.

Step S8: and bonding two silicon wafers respectively containing a micro-fluidic cavity channel and a micropore array structure together to obtain a PDMS micro-fluidic chip, and attaching the PDMS micro-fluidic chip to the glass slide 1.

The following describes the manufacturing process of the micro-pore structure and the micro-fluidic channel in detail by using specific examples, and referring to fig. 8a and 8b, the specific steps are as follows:

(1) cleaning: placing two silicon wafers in pure alcohol solution for cleaning, and drying by using nitrogen;

(2) drying: baking the cleaned silicon wafer on a hot plate at 120 ℃ for 30min to remove residual water on the silicon wafer, and cooling;

(3) gluing: placing the cleaned and dried silicon wafer on a spin coater, adding 2ml of negative photoresist SU-83025, and spin-coating at 500rpm for 15s and 2000rpm for 30 s; the photoresist is very viscous, so that the spin-coating time can be selected according to the diameter of the silicon wafer in order to improve the spin-coating yield, and the photoresist can be better spread on the silicon wafer;

(4) pre-baking: baking the spin-coated silicon wafer on a hot plate at 95 ℃ for 20min, and cooling to room temperature;

(5) exposure: respectively placing a film containing a microfluidic channel and a film with a micropore array structure right above a photoresist area on a silicon wafer, exposing the photoresist area through a photoetching machine, and setting the exposure dose of the photoetching machine to be 200mJ/cm²；

(6) Post-baking: placing the exposed silicon wafer on a hot plate at 95 ℃ to bake for 30min, and observing that the pattern is gradually shown;

(7) and (3) developing: immersing the dried and cooled silicon wafer into SU-8 developing solution, shaking the glass vessel to enable the photoresist to be fully contacted with the developing solution, flushing the soft photoresist in time for 3min, flushing the residual photoresist by spraying the developing solution, flushing the residual developing solution by using isopropanol, and drying by using nitrogen;

(8) hardening the film: baking the silicon wafer on a hot plate at 95 ℃ for 30min and cooling to volatilize water on the photoresist to obtain two silicon wafers respectively containing a microfluidic cavity channel and a micropore array structure;

(9) and (3) reversing the mold: uniformly mixing a PDMS main agent and a hardening agent according to the mass ratio of 10:1 to obtain a mixture, respectively pouring the mixture onto a silicon chip containing a micro-fluidic cavity and a micro-pore array structure, vacuumizing for 15min, and removing bubbles in PDMS;

(10) and (3) curing: placing two silicon wafers respectively containing a micro-fluidic cavity channel and a micropore array structure in an oven at 80 ℃ for baking for 1h until PDMS is completely cured;

(11) stripping: removing the cured PDMS, and punching a liquid inlet end and a liquid outlet end on a silicon chip containing the microfluidic channel by using a puncher with the hole diameter of 0.75 mm;

(12) bonding: and bonding two silicon wafers respectively containing the microfluidic cavity channel and the micropore array structure together to obtain the PDMS microfluidic chip, and attaching the PDMS microfluidic chip to the glass slide 10.

The manufacturing method of the microporous structure 50 and the cavity structure 60 provided by the embodiment of the invention is simple, low in cost and effective, and the adopted silicon chip, the photoresist and the PDMS chip are common microfluidic chip processing materials, so that the cost is low and the processing is convenient. Further, the PDMS chip may be replaced with a polymer material such as PLA, and the silicon chip may be replaced with other substrates such as polymer, glass, and silicon. The microporous structure 50 and the channel structure 60 can also be manufactured by other micro-nano processing technologies such as etching and the like.

Fig. 9 shows the adsorption and release of PS beads in the working condition of PDMS microfluidic chip: in the figure a, when the power supply is disconnected and the chip does not work, particles flow, that is, when the power supply is not available, the liquid to be separated containing the particles directly flows through the microfluidic channel 601; figure b shows the working condition of the chip that the particles are adsorbed, i.e. after the power supply is switched on, the micro bubbles are formed, and the particles in the liquid to be separated are captured and adsorbed; the graph c shows that when the power supply is turned off and the adsorbed particles are released, i.e. when the power supply is turned off again, the microbubbles disappear and the adsorbed particles are released.

Fig. 10 is a schematic diagram showing a relationship between particle size and content of a substance obtained at a sample outlet end, and shows that after a PDMS microfluidic chip works, the substance at the sample outlet end has the largest content of the substance with the particle size of 106nm under nanosight detection, a horizontal axis represents the size of the substance particle, and a vertical axis represents the content of the substance with different particle sizes, so that the substance obtained by the action of the centrifugal device provided by the present invention conforms to a particle size distribution interval of exosomes.

The centrifugal device provided by the embodiment of the invention separates the liquid to be separated through the microporous structure 50 and the cavity structure 60, namely, the liquid to be separated can be directly separated without dilution, the time consumption is low, experiments show that the exosome can be collected in only 3 hours, and the collected exosome has high concentration and purity and has cost benefit. In addition, the signal generator 20, the power amplifier 30 and the transducer 40 generate signals for excitation, namely, microbubbles are excited to resonate through external excitation, so that a microflow field is generated, the organisms in the cavity structure 60 can be controlled and screened without direct contact and damage by utilizing acoustic microflow, the magnitude of acting force can be controlled by adjusting the magnitude of input signals, the capture of substances with different micron nanometer particle sizes is realized, the device has the advantages of simple structure, high efficiency, no mark and no contact, the damage of captured substances is reduced to the maximum extent, the device has higher biological safety and higher result consistency, and also has automation and high repeatability, and the structure of the separated products is complete.

The centrifugal device provided by the embodiment of the invention can be suitable for the fields of biomedicine, chemical analysis, liquid biopsy, biochip technology and the like, can be applied to enrichment and screening of cells, microspheres and microorganisms and microfluid mixing, can also be applied to exosome extraction of biological liquids such as blood, urine, ascites, interstitial fluid, tears, saliva, cerebrospinal fluid and the like, and can realize separation of substances with different particle sizes (lipoprotein, exosome, platelet and the like) by changing energy input. The sample outlet end 8 of the centrifugal device can also be connected with a sample detection mechanism of an optical instrument, and can be used for detecting blood, urine, ascites, interstitial fluid, tears, saliva, cerebrospinal fluid and other body fluids.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims

1. A centrifuge device, characterized by: comprises a glass slide, a signal generator, a power amplifier and a transducer; the glass slide is adhered with a micropore structure and a cavity structure which are mutually bonded, and is provided with the transducer;

2. The centrifuge device of claim 1, wherein: the micropore structure is seamlessly attached to the glass slide, the cavity structure is arranged on the micropore structure, and the micropore structure and the cavity structure are bonded; and the corresponding positions of the glass slide, the micropore structure and the cavity structure are respectively provided with a positioning structure.

3. The centrifuge device of claim 2, wherein: the micropore structure comprises an equal-diameter array of micro-bubble structures, and the micro-bubbles in two adjacent rows are arranged in a staggered mode.

4. The centrifuge device of claim 3, wherein: the cavity structure comprises microfluidic cavities with symmetrical structures, all the microfluidic cavities are distributed in a snake shape, and two ends of each microfluidic cavity are respectively used as a sample inlet end and a sample outlet end.

5. The centrifuge device of claim 4, wherein: the array range of the micro-bubbles is larger than the distribution range of the micro-fluidic cavities.

6. The centrifuge device of claim 5, wherein: the micropore structure and the cavity structure are both made of PDMS chips by soft lithography and mold copying technology.

7. The centrifuge device of claim 6, wherein: the manufacturing process of the micropore structure and the cavity structure comprises the following steps:

step S6: vacuumizing the silicon wafer to remove bubbles in PDMS, and curing;

8. The centrifuge device of claim 1, wherein: the glass slide is a high-light-transmission medical glass slide.

9. The centrifuge device of claim 1, wherein: the transducer adopts a PZT piezoelectric transducer.