CN112973986B - a centrifugal device - Google Patents

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; the glass slide is attached with a micropore structure and a cavity channel structure which are bonded with each other, and the transducer is arranged on the glass slide; the cavity channel 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 micropore structure; the liquid to be separated passes through the micropore structure, forms microbubbles under the action of the ultrasonic signal and generates resonance, so as to realize the separation of different particles in the liquid to be separated. The invention has the advantages of simple and efficient structure, higher biological safety, higher result consistency, automation and high repeatability, and the product obtained by separation has complete structure.

Description

a centrifugal device

technical field

The invention relates to the technical field of biomedical engineering, and more specifically relates to a centrifugal device.

Background technique

Exosomes are a type of nanoscale vesicle structure that can be actively secreted by both normal cells and tumor cells, with a diameter of about 30-150 nm. Exosomes are widely and stably present in a variety of clinical samples, including blood, urine, ascites, tissue fluid, tears, saliva, and cerebrospinal fluid. The substances contained in exosomes from different sources are different, and exosomes provide good biological materials for the study of potential various biomarkers. Exosomes possess the specific proteins of their parent cells, which also contain various nucleic acid molecules. Therefore, various specific proteins and nucleic acid molecules carried by exosomes provide abundant detectable targets for disease analysis. Exosomes participate in the regulation of important cell physiological activities, and their roles in immune response, apoptosis, angiogenesis, inflammatory response, and coagulation have all been reported. They can be used as early diagnostic markers for various diseases and can also be used as targeted drugs carrier for disease treatment. And some exosome-based tumor therapy methods are already in clinical trials. At present, the main obstacle to the clinical application of exosomes is the lack of standard methods for rapid, stable, and efficient extraction of high-purity exosomes.

With the increasing maturity of micro-nano processing technology, microfluidic chip (also known as lab-on-a-chip, Lab-on-a-Chip) develops rapidly, in which microbubbles are widely used in microfluidic chip, such as liquid mixing (Ahmed D , Lab on a Chip, 2009, 9(18):2738-2741). The device operates by trapping air bubbles within a "horseshoe" structure located between two laminar flows inside a microchannel. After the single horseshoe-shaped structure in the cavity is immersed in the liquid, an air-liquid film is formed, and the vibration of the microbubbles generates a microflow field for liquid mixing.

Separation of different particle sizes is achieved by acoustically manipulating the chip (Mengxi W, PNAS, 201709210). This method consists of the integration of two continuous surface acoustic wave (SAW) microfluidic modules. This platform can be used directly in undiluted whole blood samples Isolate exosomes. Through the cell removal module, micro blood components with a diameter larger than 1 μm, including red blood cells, white blood cells and platelets, are extracted to obtain EV-enriched samples. Through the exosome isolation module, the removal of EVs and apoptotic bodies was achieved, and samples with purified exosomes were obtained. Each module is dependent on the tilt angle SAW field.

Existing exosome purification methods include ultracentrifugation, size-exclusion chromatography, magnetic bead-based immunoaffinity capture, polyethylene glycol-based precipitation, ultrafiltration, and microfluidics. However, the existing technology is not ideal, which limits the clinical transformation and application of exosomes. Ultracentrifugation is the most common exosome purification method, but it has low yield (recovery rate: 5%-25%), cumbersome operation process, long time-consuming (more than 1 day), and relies on expensive equipment. Due to ultra-high-speed centrifugation, it is easy to cause hemolysis and affect the experimental results. Size-exclusion-based chromatography is costly, with low recovery and low purity. The isolation method based on immune capture can collect exosomes with medium or high purity, but it is limited by the specificity of antibodies and cumbersome operation steps, it is difficult to standardize, and it is not suitable for processing large quantities and large volumes of clinical samples. The polyethylene glycol-based precipitation method is time-consuming, has many impurities, requires biomarkers, and has poor integrity of exosomes. Based on the ultrafiltration method, it is easy to clog and has low throughput. The separation method based on microfluidic technology cannot solve the problems of low copper beam, complicated operation process and poor repeatability, and it is difficult to achieve structural consistency between laboratories.

Contents of the invention

The purpose of the present invention is to solve the technical problems in the prior art, to provide a centrifugal device, which has a simple and efficient structure, and has higher biological safety and higher consistency of results, as well as automation and high repeatability. The isolated product was structurally intact.

In order to solve the problems raised above, the technical solution adopted in the present invention is:

A centrifugal device, comprising a slide glass, a signal generator, a power amplifier and a transducer; the slide glass is attached with a micropore structure and a cavity structure bonded to each other, and is provided with the transducer;

The cavity structure is used to feed the liquid to be separated; the signal generator is used to generate electrical signals, the power amplifier is used to amplify the electrical signals, and the transducer is used to convert the amplified electrical signals Generate an ultrasonic signal and act on the microporous structure; the liquid to be separated passes through the microporous structure, forms microbubbles and generates resonance under the action of the ultrasonic signal, and realizes the separation of different particles in the liquid to be separated.

Further, the microporous structure is seamlessly attached to the glass slide, the cavity structure is arranged on the microporous structure and the two are bonded; the glass slide, the microporous structure Positioning structures are respectively provided at positions corresponding to the three cavity structures.

Further, the microporous structure includes a microbubble structure in an array of equal diameters, and the microbubbles in two adjacent rows are arranged in a staggered manner.

Further, the channel structure includes microfluidic channels with a symmetrical structure, all of the microfluidic channels are distributed in a serpentine shape, and the two ends of the channels are respectively used as sample inlet ports and sample outlet ports.

Further, the array range of the microbubbles is larger than the distribution range of the microfluidic channel.

Further, both the microporous structure and the cavity structure are made of PDMS chips, and are manufactured by soft lithography and mold printing techniques.

Further, the manufacturing process of the microporous structure and the channel structure includes the following steps:

Step S1: Select two silicon wafers and place them in pure alcohol solution to clean them, dry them with nitrogen, and place them on a hot plate to bake and cool;

Step S2: Place the cleaned and dried silicon wafer on a spin coater, add a negative photoresist for spin coating, and place it on a hot plate to bake and cool;

Step S3: Place the film sheet containing the microfluidic cavity and the film sheet containing the microhole array structure directly above the photoresist area on the silicon wafer, and perform photolithography on the photoresist area by a photolithography machine Exposure treatment, and placed on a hot plate to bake and cool;

Step S4: immerse the exposed silicon wafer in the SU-8 developer, shake the glass dish, spray the developer and rinse with isopropanol, dry it with nitrogen, and place it on a hot plate to bake and cool. Obtain two silicon wafers respectively containing microfluidic cavity and microhole array structure;

Step S5: mixing the PDMS main agent and the hardening agent in proportion to obtain a mixture, and pouring the mixture into the silicon wafer containing the microfluidic cavity and the silicon wafer containing the microwell array structure;

Step S6: vacuumize the silicon wafer to remove the air bubbles in the PDMS, and perform curing treatment;

Step S7: peel off the cured PDMS, and use a puncher to punch holes on the silicon wafer containing the microfluidic cavity as the inlet and outlet of the liquid to be tested;

Step S8: bonding two silicon wafers respectively containing the microfluidic cavity and the microwell array structure together to obtain a PDMS microfluidic chip, and attaching it to the glass slide.

Further, the slides are high-transmittance medical slides.

Further, the transducer adopts a PZT piezoelectric transducer.

Compared with prior art, the beneficial effect of the present invention is:

The invention directly separates the liquid to be separated through the microporous structure and the cavity structure without dilution, and generates excitation signals through the signal generator, power amplifier and transducer to act on the microbubbles to generate resonance, so that there is no direct contact and no damage. Manipulation and screening, and real-time centrifugation of various gradient nano-micron-level substances by adjusting different power energies. It has the characteristics of simple structure, low input energy, low cost, and high timeliness. It can also be directly obtained from biological fluids on a single device in an automatic way Separation of the required substances, the isolated substances have high purity, high yield and complete structure.

Description of drawings

Figure 1 is a schematic diagram of the overall structure of the centrifugal device of the present invention.

Fig. 2 is a plane structure diagram of a glass slide in the present invention.

Fig. 3 is a plan view of the microporous structure in the present invention.

Fig. 4 is a plan view of the lumen structure of the present invention.

Fig. 5 is a schematic diagram of the microbubble structure in the present invention.

Fig. 6 is a conceptual diagram of cells captured by array microbubbles in the present invention.

Fig. 7 is a flow chart of the fabrication of the microporous structure and the cavity structure in the present invention.

Fig. 8 is a schematic diagram of the fabrication of the microporous structure and the cavity structure in the present invention.

Fig. 9 is the situation of adsorption and release of PS pellets in the chip working condition in the present invention.

Fig. 10 is a schematic diagram of the relationship between the particle size of the material obtained at the outlet of the sample and its content in the present invention.

The description of the drawings is as follows: 10-slide, 20-signal generator, 30-power amplifier, 40-transducer, 50-micropore structure, 60-cavity 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, 601-microfluidic cavity.

Detailed ways

In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the associated drawings. Preferred embodiments of the invention are shown in the accompanying drawings. However, the present invention can be embodied in many different forms and is not limited to the embodiments described herein. On the contrary, these embodiments are provided to make the understanding of the disclosure of the present invention more thorough and comprehensive.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the technical field of the invention. The terms used herein in the description of the present invention are for the purpose of describing specific embodiments only, and are not intended to limit the present invention.

Referring to Fig. 1, an embodiment of the present invention provides a centrifugal device, including a slide glass 10, a signal generator 20, a power amplifier 30, and a transducer 40; the slide glass 10 is bonded with the micropore The structure 50 and the channel structure 60 are provided with the transducer 40; the microporous structure 50 and the channel structure 60 are bonded.

The cavity structure 60 is used to feed the liquid to be separated; the signal generator 20 is used to generate an electrical signal, the power amplifier 30 is used to amplify the electrical signal, and the transducer 40 is used to amplify the electrical signal. The amplified electrical signal is converted into an ultrasonic signal and acts on the microporous structure 50; the liquid to be separated passes through the microporous structure 50, forms microbubbles and generates resonance under the action of the ultrasonic signal, and realizes separation Separation of different particles in a liquid.

In the centrifuge device provided by the embodiment of the present invention, after the cavity structure 60 is fed into the liquid to be separated by the external low-pressure peristaltic pump, due to the surface tension of the liquid, after the liquid to be separated flows through the microporous structure 50, the ultrasonic signal Under the excitation of , a liquid-air film is formed and microbubbles are formed. There is a pressure difference between the air surrounded by the liquid to be separated and the surrounding liquid, which makes the formed microbubbles produce steady-state cavitation, and resonance occurs to realize the difference in the liquid to be separated. Separation of particles. Since the resonant frequency of the microbubbles depends on the magnitude of the input energy, in the embodiment of the present invention, the vibration amplitude of the microbubbles can be changed by adjusting the energy or frequency of the electrical signal input by the signal generator 20, so as to realize the vibration of different particles. capture, and the resonance of the microbubble can be changed by adjusting different amplitudes of the signal generator 20 to achieve separation of different nanometer or micrometer particle sizes.

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

In the embodiment of the present invention, the microporous structure 50 includes microbubble structures 501 distributed in an array of equal diameters, and the microbubble structures 501 in two adjacent rows are arranged in a staggered manner (see FIG. 5 ), which can realize uniform distribution of substances in the liquid. In the microfluidic cavity, the large substances are sufficiently and evenly captured by the air bubbles, thereby reliably realizing large-scale particle screening. Further, the diameter of the microbubble structure 501 is preferably 40 μm. Since microbubbles with different diameters have different vibration amplitudes, the disturbance to the surrounding environment is also different, and the speed of particles with different diameters in the same microbubble field is also Therefore, particles with different diameters can be screened by changing the diameter of the microbubble structure 501 .

In the embodiment of the present invention, the channel structure 60 includes microfluidic channels 601 with a symmetrical structure, and all the microfluidic channels 601 are distributed in a serpentine shape, and the two ends thereof are respectively used as the sample inlet port 2 and the sample outlet port 8 . The microfluidic channel 601 adopts a symmetrical structure and is distributed in a serpentine shape, so that when the liquid is introduced, the liquid can fully filter out larger substances in the microfluidic channel 601 to ensure the reliability of separation. The microfluidic cavity 601 can hold blood, urine or other liquids to be separated.

Further, the two ends of the microfluidic channel 601 are perforated by a puncher as the sample inlet port 2 and the sample outlet port 8, and the liquid to be separated is injected into the sample inlet port 1 through a hose through a syringe. In the microfluidic cavity 601, it can ensure that the whole separation process is safe, reliable and free from pollution.

In the embodiment of the present invention, the array range of the microbubbles 501 on the microporous structure 50 is larger than the distribution range of the microfluidic channels 601, which facilitates reliable bonding between the two.

In the embodiment of the present invention, the slide glass 10 is a high-transmittance medical slide glass, which has a good effect.

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

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

After the microfluidic cavity 601 is injected with the liquid to be separated, stable microbubbles are formed when the liquid to be separated passes through the microbubble structure 501 of the array. Vibration occurs to generate micro-flow. The surrounding fluid medium is affected by the "breathing" effect of the surface membrane, and the energy is transferred to the particles (ie, the substances to be separated) in the microfluidic cavity 601 to realize the manipulation of the particles. The particles in the microfluidic cavity 601 are mainly affected by the acoustic radiation force and the microfluidic induced pulling force under the vibration of the microbubbles, and the incident waves generated by the resonant microbubbles are scattered, causing the particles to be affected by the radiation force. The effect, that is, the force generated by the microbubbles can adsorb the particles in the liquid to be separated. The distance between the particles and the microbubbles in the microfluidic cavity 601 has a great influence on the magnitude of the acoustic radiation force. 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 cavity 601 is lower than that of the liquid, they are repelled. When the density of the liquid is equal to that of the particle, the radiation force experienced by the particle is very small. The microfluidic radiation force increases with the increase of the microbubble diameter, and the particle diameter is also positively correlated with the radiation force.

In the embodiment of the present invention, the microporous structure 50 and the cavity structure 60 are both made of PDMS (polydimethylsiloxane: polydimethylsiloxane) chips, and are produced by soft lithography and mold printing technology, refer to the figure 7, the specific production process includes the following steps:

Step S1: Select two silicon wafers, clean them in pure alcohol solution, blow them dry with nitrogen, and place them on a hot plate to bake and cool.

Step S2: Place the cleaned and dried silicon wafer on a spin coater, add a negative photoresist for spin coating, and place it on a hot plate to bake and cool.

Step S3: Place the film sheet containing the microfluidic cavity and the film sheet containing the microhole array structure directly above the photoresist area on the silicon wafer, and perform photolithography on the photoresist area by a photolithography machine Expose and place on a hot plate to bake and cool.

Step S4: immerse the exposed silicon wafer in the SU-8 developer, shake the glass dish, spray and rinse the residual photoresist with the developer, rinse with isopropanol to remove the residual developer, and blow dry with nitrogen. And placed on a hot plate to bake and cool, to obtain two silicon wafers respectively containing microfluidic cavity and microhole array structure.

Step S5: mixing the PDMS main agent and the hardener in proportion to obtain a mixture, and pouring the mixture onto the silicon wafer containing the microfluidic cavity and the silicon wafer containing the microwell array structure respectively.

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

Step S7: peel off the cured PDMS, and use a hole puncher to punch holes on the silicon wafer containing the microfluidic cavity as the inlet port and the outlet port of the liquid to be tested.

Step S8: bonding two silicon wafers respectively containing the microfluidic cavity and the microwell array structure together to obtain a PDMS microfluidic chip, and attaching it to the slide glass 1 .

The manufacturing process of the microporous structure and the microfluidic channel will be described in detail below through specific examples, as shown in Figure 8a and Figure 8b, and the specific steps are as follows:

(1) Cleaning: Clean the two silicon wafers in pure alcohol solution, and dry them with nitrogen;

(2) Drying: bake the cleaned silicon wafer on a hot plate at 120°C for 30 minutes to remove residual moisture on the silicon wafer and cool it;

(3) Coating: place the clean and dried silicon wafer on a spin coater, add 2ml of negative photoresist SU-8 3025, and spin coat at 500rpm for 15s and 2000rpm for 30s; The resist is very viscous. In order to improve the spin-coating yield, the spin-coating time can be selected according to the diameter of the silicon wafer, so that the photoresist can be better spread on the silicon wafer;

(4) Pre-baking: bake the spin-coated silicon wafer on a hot plate at 95°C for 20 minutes, and cool to room temperature;

(5) Exposure: Place the film sheet containing the microfluidic cavity and the film sheet with the microwell array structure directly above the photoresist area on the silicon wafer, and expose the photoresist area through a photolithography machine, and set The exposure dose of the lithography machine is 200mJ/cm ² ;

(6) Post-baking: place the exposed silicon wafer on a hot plate at 95°C and bake for 30 minutes, and the pattern can be observed to gradually appear;

(7) Development: Immerse the dried and cooled silicon wafer in the SU-8 developer, shake the glass dish, make the photoresist fully contact with the developer, wash away the soft photoresist in time, last for 3min, and use the developer Spray and rinse the residual photoresist, rinse with isopropanol to remove the residual developer, and then blow dry with nitrogen;

(8) Hardening film: put the silicon wafer on a hot plate at 95°C for 30 minutes and cool it down to volatilize the water on the photoresist to obtain two silicon wafers containing microfluidic cavity and micropore array structure respectively. piece;

(9) Inverting the mold: mix the PDMS main agent and the hardening agent evenly at a mass ratio of 10:1 to obtain a mixture, and pour the mixture into the silicon alloy containing the microfluidic cavity and the micropore array structure. On the chip, vacuumize for 15 minutes to remove the air bubbles in PDMS;

(10) Curing: Place two silicon wafers containing microfluidic cavity and micropore array structure respectively in an oven at 80°C and bake for 1 hour until the PDMS is completely cured;

(11) Peeling: peel off the cured PDMS, and use a hole puncher with an aperture of 0.75 mm to punch holes at the liquid inlet and outlet ports on the silicon wafer containing the microfluidic cavity;

(12) Bonding: bonding two silicon wafers respectively containing the microfluidic cavity and the microwell array structure together to obtain a PDMS microfluidic chip, and the PDMS microfluidic chip is bonded on the glass slide 10 .

The manufacturing method of the microporous structure 50 and the cavity structure 60 provided by the embodiment of the present invention is simple, low-cost and effective, and the silicon wafer, photoresist and PDMS chip used are all common microfluidic chips Processing materials, low cost and convenient processing. Further, the PDMS chip can also be replaced by polymer materials such as PLA, and the silicon chip can also be replaced by other polymers, glass, silicon and other substrates. The microporous structure 50 and the channel structure 60 can also be fabricated by other micro-nano processing techniques such as etching.

Figure 9 shows the adsorption and release of PS beads when the PDMS microfluidic chip is working: Among them, Figure a shows the flow of particles when the chip is not working when the power is turned off, that is, in the absence of power, the liquid to be separated containing particles It will flow directly through the microfluidic cavity 601; Figure b shows that the particles are adsorbed when the chip is working, that is, after the power is turned on, microbubbles are formed, and the particles in the liquid to be separated are captured and adsorbed; Figure c is the adsorption when the power is turned off. The particles are released, that is, when the power is disconnected again, the microbubbles disappear and the adsorbed particles are released.

Figure 10 is a schematic diagram of the relationship between the particle size of the substance obtained at the sample outlet and its content. It shows that after the PDMS microfluidic chip works, the substance at the sample outlet is detected by nanosight, and the particle size of the substance is 106nm. The axis is the particle size of the substance, and the vertical axis is the content of substances with different particle sizes, so it shows that through the action of the centrifugal device provided by the present invention, the obtained substances conform to the particle size distribution range of exosomes.

The centrifuge device provided by the embodiment of the present invention separates the liquid to be separated through the microporous structure 50 and the cavity structure 60, that is, it can be directly separated without dilution, and it takes less time. Experiments show that it only takes 3 hours. Exosomes are collected, and the collected exosomes are high in concentration, high in purity, and cost-effective. In addition, the signals generated by the signal generator 20, the power amplifier 30, and the transducer 40 are used for excitation, that is, the resonance is generated by externally exciting the microbubbles, thereby generating a microfluidic field. Carry out no direct contact, no damage manipulation and screening, and can control the size of the force by adjusting the size of the input signal, so as to realize the capture of substances with different micron and nanometer particle sizes. It can greatly reduce the damage of captured substances, and has higher biological safety and higher consistency of results. It also has automation and high repeatability, and the structure of the separated product is complete.

The centrifugal device provided by the embodiment of the present invention can be applied to the fields of biomedicine, chemical analysis, liquid biopsy, biochip technology, etc. It can be used in the enrichment and screening of cells, microspheres, microorganisms, and microfluidic mixing. Extract exosomes from biological fluids such as blood, urine, ascites, tissue fluid, tears, saliva, and cerebrospinal fluid, and can separate substances of different particle sizes (lipoproteins, exosomes, platelets, etc.) by changing the energy input . The sample outlet port 8 of the centrifugal device can also be connected to an optical instrument sample detection mechanism, which can be used to detect blood, urine, ascites, interstitial fluid, tears, saliva and cerebrospinal fluid and other body fluids.

The above-mentioned embodiment is a preferred embodiment of the present invention, but the embodiment of the present invention is not limited by the above-mentioned embodiment, and any other changes, modifications, substitutions, combinations, Simplifications should be equivalent replacement methods, and all are included in the protection scope of the present invention.

Claims (7)

1. A centrifugal device, characterized in that: comprises a glass slide, a signal generator, a power amplifier and a transducer; the glass slide is attached with a micropore structure and a cavity channel structure which are bonded with each other, and the transducer is arranged on the glass slide; the micropore structure comprises a microbubble structure of an equal-diameter array, and two adjacent rows of microbubble structures are arranged in a staggered manner; the cavity structure comprises micro-fluidic cavities with symmetrical structures, all the micro-fluidic cavities adopt serpentine distribution, and two ends of the micro-fluidic cavities are respectively used as a sample inlet end and a sample outlet end;

the cavity channel 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 micropore structure; the liquid to be separated passes through the micropore structure, forms microbubbles under the action of the ultrasonic signal and generates resonance, so as to realize the separation of different particles in the liquid to be separated.

2. A centrifugal device according to claim 1, wherein: the microporous structure is in seamless fit with the glass slide, and the cavity channel structure is arranged on the microporous structure and is bonded with the microporous structure; and positioning structures are respectively arranged at the corresponding positions of the glass slide, the micropore structure and the cavity channel structure.

3. A centrifugal device according to claim 1, wherein: the array range of the microbubbles is larger than the distribution range of the microfluidic channels.

4. A centrifugal device according to claim 3, wherein: the micropore structure and the cavity channel structure are both made of PDMS chips and are manufactured through soft lithography and die copying technologies.

5. The centrifuge apparatus of claim 4 wherein: the manufacturing process of the micropore structure and the cavity channel structure comprises the following steps:

step S1: two silicon chips are selected, placed in pure alcohol solution for cleaning, dried by nitrogen, and placed 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 on a hot plate for baking and cooling;

step S3: respectively placing a film containing a microfluidic cavity and a film containing a micropore array structure right above a photoresist area on the silicon wafer, exposing the photoresist area through a photoetching machine, and placing the film and the film on a hot plate for baking and cooling;

step S4: immersing the exposed silicon wafer into SU-8 developing solution, shaking the glass dish, spraying the developing solution and flushing with isopropanol, blow-drying with nitrogen, and placing on a hot plate for baking and cooling to obtain two silicon wafers respectively containing microfluidic channels and micropore array structures;

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

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

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

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

6. A centrifugal device according to claim 1, wherein: the glass carrier chip is a high-light-transmission medical glass carrier.

7. A centrifugal device according to claim 1, wherein: the transducer is a PZT piezoelectric transducer.

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