CN114621857A

CN114621857A - High-flux cracking system based on resonance microbubble array

Info

Publication number: CN114621857A
Application number: CN202011440596.9A
Authority: CN
Inventors: 郑海荣; 刘秀芳; 孟龙; 徐礼胜; 荣宁; 牛丽丽
Original assignee: Shenzhen Institute of Advanced Technology of CAS
Current assignee: Shenzhen Institute of Advanced Technology of CAS
Priority date: 2020-12-10
Filing date: 2020-12-10
Publication date: 2022-06-14

Abstract

The invention relates to a high flux cracking system based on a resonance microbubble array, in particular to a device for cracking, which comprises a shell, a first heating device, a second heating device, a first heating device and a second heating device, wherein the shell is provided with a first heating chamber and a second heating chamber; 1) a cavity body of the micro-bubble array is formed, the cavity body is provided with a fluid cavity channel, and two sides of the fluid cavity channel are provided with micro-structure cavity bodies which are communicated with the cavity channel and protrude outwards, and the micro-structure cavity bodies can capture and keep micro-bubbles; 2) the substrate is bonded with the cavity, and a fluid channel capable of enabling fluid to pass is formed between the substrate and the fluid channel of the cavity together; 3) the body wave generating device can generate body waves and transmit the body waves into the cavity forming the microbubble array to form resonance with the microbubbles. The device of the invention has simple structure and can realize the lysis of cells, bacteria, viruses or tissues with high flux.

Description

High-flux cracking system based on resonance microbubble array

Technical Field

The invention belongs to the field of molecular biology, and particularly relates to a system for rapidly cracking cells or viruses based on a resonance microbubble array.

Background

In recent years, cell lysis has attracted much attention as the separation and purification of cellular components have become important in disease diagnosis, gene therapy, cell engineering and drug screening. Cell lysis is a process by which the nuclear membrane is disrupted to release intracellular material (e.g., DNA, RNA, proteins or organelles). However, the development of an efficient and rapid cell lysis technique is crucial for the isolation and purification of intracellular components. Currently, common cell lysis techniques fall into two broad categories: mechanical and non-mechanical cracking processes. Although the mechanical cracking method can achieve high-flux cracking effect, the method is easy to generate strong thermal effect and is not suitable for extracting proteins in cells. In contrast, non-mechanical cell lysis techniques are more popular. Biological and chemical non-mechanical lysis technology can prevent mechanical damage to intracellular components, is particularly suitable for mechanically sensitive intracellular components, and has the defects of expensive reagents, complex operation and the like. Physical cell lysis techniques are widely used in experiments and in practical production as the most promising method among non-mechanical cell lysis techniques. Currently, the most commonly used non-mechanical cell lysis techniques are biological lysis techniques, chemical lysis techniques, temperature lysis techniques, osmotic fracturing lysis techniques, photolysis techniques, electrical lysis techniques and acoustic lysis techniques.

The biological cracking technology comprises the following steps: enzyme lysis techniques one commonly used enzyme lysis is a biological cell lysis method, the variety of enzymes is large, specific types of cells require specific lytic enzymes, e.g., lysozyme for bacterial cell lysis, chitinase for yeast cell lysis, and pectinase for plant cell lysis. The greatest advantage of enzymatic cleavage is specificity. However, the specificity of the enzyme limits its limitations, which is not suitable for wide application. And there is a problem that the cells cannot be completely lysed. ([1] Andrews, B.A.; Asenjo, J.A.enzymic lysis and displacement of microbial cells T references Biotechnol.1987,5, 273- & lt 2 & gt Salazar, O.; Asenjo, J.A.enzymic lysis of microbial cells Biotechnol.Lett.2007,29, 985- & lt 994.)

Chemical cracking technology: the cell membrane is disrupted by changing the pH of the chemical lysis solution. Surfactants may also be added to the cell lysis buffer to solubilize membrane proteins to disrupt the cell membrane and release its contents. Chemical cracking techniques can be divided into alkaline cracking and surfactant cracking. In alkaline cleavage techniques, OH^﹣Ions are the main component of the lysis of cell membranes. Lysis buffer consisted of sodium hydroxide and Sodium Dodecyl Sulfate (SDS). OH group^﹣The fatty acid-glycerol ester bond in the cell membrane can be destroyed and then the cell membrane is made permeable, and SDS dissolves proteins and membranes. The pH range of the lysate is generally controlled to be 11.5-12.5. In the surfactant cleavage technique, surfactants can disrupt the double lipid layer consisting of hydrophobic and hydrophilic molecules in the cell membrane, mainly by disrupting the lipid-lipid, lipid-protein and protein-protein interactions in the cell membrane. Although this method is applicable to all types of cells, the cell lysis process is very slow, requiring about 6 to 12 hours. ([3]Stanbury,P.F.；Whitaker,A.Principles of Fermentation T echnology；Pergamon Press:Oxford,UK,1984.[4]Feliciello,I.；Chinali,G.A modified alkaline lysis method for the preparation of highly purified plasmid DNA from Escherichia coli.Anal.Biochem.1993,212,394–401.)

Temperature cracking technology: is a technique that causes cell lysis by repeatedly freezing and thawing cells, resulting in the formation of ice crystals on the cell membrane, causing the cell membrane to disintegrate. At the same time, high temperatures can also damage membranes by denaturation of cell membrane proteins and lead to release of intracellular organelles. However, this lysis technique, while not limited to cell types and easy to implement, is time consuming, expensive and cannot be used to extract temperature sensitive intracellular components. ([5] Johnson, B.H.; Hecht, M.H.Cellsby reproduced cycles of fresh and thawing.Biotechnology 1994,12, 1357; Zhu, K. [6] Jin, H.; He, Z.; Zhu, Q.; Wang, B.Acontinuus method for the large-scale extraction of plasma DNA by modified manufacturing lysine.Nat.Protoc.2007, 1, 3088-

And (3) osmotic fracturing solution technology: when the extracellular fluid concentration is lower than the intracellular fluid concentration, the extracellular fluid enters the cells due to osmosis, which in turn causes the cells to swell and then rupture. The cell membrane structure of mammals is fragile, so the technology is suitable for cracking the cells of mammals and can be used for extracting sensitive components in the cells, but the technology is not suitable for all types of cells, and therefore, the application of the cracking technology is limited to a great extent. ([7] Fonseca, L.P.; Cabral, J.Penicillin acylase release from Escherichia coli cells by mechanical cell dispersion and permeabilization.J.Chem.T.Echnol.Biotechnol.2002, 77,159 167.[8] Chen, Y. -C.; Chen, L. -A.; Chen, S. -J.; Chang, M. -C.; Chen, T. -L.Amodified acyl synthase for epidermal release of an immunogenic cleavage from Escherichia coli. Biochem.Eng.J.2004,19, 211-

The photocleavage technology comprises the following steps: the cell is controlled by utilizing laser targeting, after laser irradiation, the focused laser pulse at the interface of the cell solution can generate cavitation bubbles, and then the high-intensity shock wave generated by the cavitation of the bubbles can cause cell lysis through the laser irradiation. The technology is a non-contact, single-cell operation, high-efficiency, applicable to various cells, in-vivo research, sterile environment and applicable to a lab-on-a-chip, but in the process of researching cell lysis, the requirement on the number of cells is strict, required equipment is expensive, the operation is complex, the time required for lysis is long, and the technology is not beneficial to wide application. ([9] Huang, S. -H.; Hung, L. -Y.; Lee, G. -B.B.continuous nuclear expression by optical synthesis-induced cell lysis on a batch-type microfluidic platform, Lab Chip 2016,16, 1447-

The electric cracking technology comprises the following steps: the high-voltage electric field acts on a phospholipid bilayer of a cell membrane in microsecond and millisecond pulse mode to generate unstable potential, high-strength transmembrane potential is formed in the cell, and high-strength potential is generated to cause cell lysis. The technology is high in cell lysis efficiency and short in time consumption, but the requirement on experimental conditions is high, equipment is expensive, the heat production amount is high, and the technology is not beneficial to extracting temperature-sensitive intracellular components, so that the wide application of the technology is limited to a great extent. ([11] Ohshima, T.; Sato, M.; Saito, M.Selective release of interstitial protein using pulsed electronic field.J.Electrost.1995,35, 103- & 112.[12] America, S.K.; Singh, P.K.; Dokmeci, M.R.; Khadeshoseini, A; Xu, Q.; Sonkusal, S.R.All electronic approach for high-through cell bridging and lysine with electronic impedance monitoring & biosensing.Biosens.Bioelectronic 2014.54, 462, 467.)

The acoustic cracking technology comprises the following steps: the cell lysis technology is realized by means of the combination of sound wave energy and the cavitation effect of micro bubbles. Under the action of ultrasound, the microbubbles oscillate near the biological wall to generate microjets around the biological wall, and the shearing force generated by the microjets on the biological wall can rupture cell membranes. The method is a novel method for separating and purifying intracellular components by an ultrasonic cell lysis technology, the realization of the ultrasonic cell lysis technology mainly depends on the cavitation effect of the microbubbles, however, the sizes and resonance frequencies of the microbubbles are not uniform, and the randomness of the cavitation effect is strong; the application and development of the acoustic lysis technology have some obstacles due to different cell sizes and acceptable shearing force thresholds of cells, and the like, which are acted with the microbubbles. Due to the difference of ultrasonic radiation systems, radiation dose calculation methods have diversity, no relatively determined ultrasonic dose for cell lysis exists at present, and some parameters in the ultrasonic radiation effect cannot be accurately controlled. Meanwhile, the cell lysis of the acoustic lysis technology in the current stage also has the problems of high heat production and low cell lysis efficiency. ([12] Lenticker I, De Cock I, Decker R.et al.outstanding interaction resources: Definitions and uncovering mechanisms [ J ]. Advanced Drug Delivery reviews.2014,72:49.[14] Study of a novel cell approach with titanium dioxide for lab-on-a-chip device biomed.microdevice 2011,13, 527- & 532.)

Disclosure of Invention

Aiming at the defects, the invention provides a micro-fluidic chip cell lysis system based on a resonance microbubble array, and provides a micro, single-cell, high-throughput, rapid and efficient cell lysis system which has extremely low heat production, simple operation and high repeatability, can be suitable for extracting and purifying any component in cells, and can be widely applied to various types of cells. The micro-fluidic system without cell and cell component limitation realizes rapid, efficient and high-stability cell lysis by combining microbubbles with uniform sizes through ultrasonic waves based on the steady-state cavitation effect of the microbubbles, and shows application prospects in disease diagnosis, gene therapy, cell engineering and drug screening.

One aspect of the invention provides a device for lysing cells, the device comprising;

1) forming a cavity of a micro-bubble array, wherein the cavity is provided with a fluid cavity, and two sides of the fluid cavity are provided with micro-structure cavities which are communicated with the cavity and protrude outwards, and the micro-structure cavities can capture and hold micro-bubbles;

2) the substrate is bonded with the cavity, and a fluid channel capable of enabling fluid to pass is formed between the substrate and the fluid channel of the cavity together;

3) the body wave generating device can generate body waves, transmit the body waves into the cavity forming the microbubble array and form resonance with the microbubbles;

in some embodiments of the invention, the bulk wave generating device is a bulk wave transducer. In a preferred embodiment, the frequency of the bulk wave transducer is 100-120kHz, preferably 105-110kHz, and in a specific embodiment 107kHz is used.

In some embodiments of the invention, the apparatus further comprises a signal generator and a power amplifier.

In some embodiments of the invention, the signal generator is configured to generate a sine wave signal and to send the sine wave signal to the power amplifier.

In some embodiments of the invention, the power amplifier is configured to amplify the sine wave signal and send the amplified sine wave signal to the bulk wave transducer.

In some embodiments of the invention, the cavity forming the array of microbubbles has one or more fluid channels thereon, and the microstructure cavities are staggered on both sides of the channels.

In some embodiments of the invention, the microstructure cavities have dimensions of 1-100 μm, preferably 20-60 μm in width and height; preferably, the width and height of the microstructure cavity is 0.8-1.2 times the diameter of the cell to be lysed. In a preferred embodiment of the invention, the microstructure cavities have a width of 40.8 μm and a height of 50 μm. In the technical scheme of the invention, the thickness of the microstructure cavity is higher than the cell diameter and less than 5 cm.

In some embodiments of the invention, the cavity and the substrate are plasma bonded.

In some embodiments of the invention, the material of the cavity is a siloxane, preferably Polydimethylsiloxane (PDMS).

In some embodiments of the invention, the cavity in which the array of microbubbles is formed has at least one inlet and one outlet. In some preferred embodiments of the invention, the inlet is in communication with an inlet means and the outlet is in communication with a cell lysate recovery means. In some preferred embodiments of the present invention, the liquid feeding device is a micro-syringe pump. In some preferred embodiments of the invention, the cell lysate recovery apparatus is an analytical apparatus, such as a PCR device, a cell analysis device, or the like.

In another aspect, the invention provides a method for preparing the above device, wherein the cavity for forming the microbubble array is obtained by photolithography.

In a further aspect, the present invention provides a method for lysing cells, viruses, bacteria or tissues, which employs the above-described apparatus for lysing cells according to the present invention and comprises the steps of:

1) placing a solution containing cells, viruses, bacteria or tissues in a lumen of the device, wherein the micro-structure cavity of the lumen captures and generates microbubbles;

2) the body wave is generated by a body wave generating device in the microfluidic device, so that the microbubbles resonate to cause the vibration of the solution in the cavity, and a flow field is generated in the cavity for cracking cells or viruses.

In some embodiments of the invention, the solution comprising cells, viruses, bacteria or tissues in step 2) is retained in the flow field generated by the body waves for at least 3 minutes.

In some embodiments of the invention, further comprising step 3) recovering the lysed cells or virus fluid.

In the technical scheme of the invention, the shear stress in the flow field at the periphery of the micro-bubble is shown as follows

S＝2π^3/2ε²(ρ_ff³μ)^1/2/R_b，

Wherein R is_hIs the radius of the microbubble, f is the resonance frequency of the microbubble, ρ_fWhere the density of the liquid, μ is the relative fluid velocity and ε is the amplitude of the oscillation of the microbubbles.

In a further aspect the invention provides the use of the device of the invention as described above for cell lysis, bacterial lysis, viral lysis or tissue understanding.

Advantageous effects

(1) The device of the present invention is primarily characterized by high throughput, and the device of the present invention can be used to fabricate multiple channels, in one embodiment three parallel rows of channels are fabricated. The micro-structure cavities are arranged on the two sides of the cavity, flow fields generated by different micro-bubbles are mutually interacted, and special flow field force exists between independent flow fields, so that the cracking efficiency of cells, viruses, bacteria or tissues can be improved, and the vibration generated by the micro-bubbles in the micro-structure cavities can act on the cells in the cavity to the maximum extent.

(2) The micro-structure cavity can capture micro-bubbles with the same radius, the array micro-bubbles vibrate simultaneously under the excitation of the body wave generating device, the amplitude is the same, experiments verify that the micro-bubble vibration is a steady-state cavitation process, and the generated equivalent second-order acoustic radiation force can capture cells around the micro-bubbles in the solution on the surfaces of the micro-bubbles. The cells are lysed by the generation of an equivalent shear stress. Due to the generation of equivalent shear stress, the cell lysis efficiency is greatly improved, and larger cell sample amount can be processed in the same time.

(3) The precision is that the cells are captured on the surfaces of the microbubbles through the second-order acoustic radiation force, and the distance between the cells and the flow field can be controlled by the body waves, so that the shearing stress on the cells is accurately controlled, and the cell lysis efficiency is accurately controlled. The invention realizes the control of the cell position by only using the body wave generating component without using other components for controlling the cell position, captures the cells on the surface of the micro-bubbles and realizes high-efficiency lysis.

(4) The device has repeatability, the cavity channel can be processed by a standard MEMS process, the device has good consistency, and the repeatability of the experiment is further improved.

(5) The lysis method has the characteristics of rapidness and simple operation, the understanding of cells can be realized only by using a commercially available cell lysis kit within 30 minutes, but the cell lysis can be completed only within 3 minutes by using the device disclosed by the invention, and continuous and uninterrupted cell lysis can be realized after an outlet and an inlet of the device are respectively coupled with a liquid inlet device and a cell lysate recovery device, so that the number of cells on a substrate can be increased, the number of cells lysed simultaneously can also be increased, and the lysis efficiency can be increased by geometric times compared with that of the commercial kit. In addition, the cracking method of the invention can ensure high efficiency and simultaneously realize no damage to DNA and protein. For example, at an input voltage of 144Vpp, the lysis efficiency of MCF-7 cells can reach 97.6 percent by ultrasonic action for 1 min. And only 5min is needed for manufacturing a cavity channel of PDMS, the structural template of PDMS can be recycled, and only 10min is needed for building the whole experiment platform.

(6) The invention utilizes ultrasonic wave to regulate cell lysis, and the cell type is universal. It is only necessary to involve different channels and microstructure cavities depending on the size of the cells.

(7) The method has the characteristic of extremely low heat generation, and can be used for extracting and analyzing temperature-sensitive protein or enzyme.

Drawings

FIG. 1 is a flow chart of the preparation of PDMS channels.

FIG. 2 is a diagram of a PDMS channel structure.

FIG. 3 is a schematic structural diagram of the experimental apparatus.

FIG. 4 is a flow field diagram generated by resonance of micro bubbles at PDMS micro-holes.

FIG. 5 shows the effect of cell lysis in this system, which is observed by Propidium Iodide (PI) and Calcein (Calcein-AM) double staining method in combination with fluorescence microscope.

Detailed Description

In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, specific embodiments thereof are described in detail below, but the present invention is not to be construed as being limited to the implementable range thereof.

Example 1 PDMS Cavity preparation and bonding

The process of making PDMS is shown in FIGS. 1 (a-e).

(1) Pretreatment: removing residual impurities such as dust, organic adsorbates and the like on the surface of the silicon wafer by acid washing, alcohol washing, water washing and other methods, and finally placing the silicon wafer in a clean place for airing.

(2) Gluing and pre-drying: the coating is spin-coated on SU-8(50) negative photoresist with the thickness of about 50 μm at 3000rpm for 30s and SU-8 (50). After the photoresist is coated, the silicon wafer is horizontally placed on a heating plate at 90 ℃ for 1h, so that the solvent in the photoresist is volatilized, the adhesion between the photoresist and the silicon wafer is enhanced, and the silicon wafer with the photoresist layer on the surface is obtained, wherein the structure is shown as (a) in fig. 1.

(3) Exposure and development: and (b) placing a film sheet (shown as fig. 1) with a pattern on the side with the photoresist layer of the silicon wafer obtained in the step (2), wherein the film sheet is provided with a hollowed-out pattern, and the hollowed-out pattern is shown as fig. 2. Exposing with an exposure machine at a dose of 600cJ/cm²For a duration of 30 s. Soaking the exposed silicon wafer with developing solution to dissolve the photoresist layer in the unexposed region and retain the photoresist layer in the exposed region, and developingAnd then baking the silicon wafer on a heating plate at 150 ℃ for 10min to obtain the photoresist layer adhered to the silicon wafer, wherein the shape of the photoresist layer is as shown in the figure 1 (d).

(4) Casting PDMS: and (3) proportioning the glue A and the glue B of the PDMS according to the mass ratio of 10:1, uniformly mixing, putting the mixture into a culture dish in which the silicon wafer obtained in the step (3) is positioned, vacuumizing the culture dish, removing microbubbles in the PDMS, finally putting the culture dish in an oven at 80 ℃ for 1h, and curing the PDMS to obtain a product with the structure shown in (e) in figure 1, wherein the shape of the photoresist layer forms a negative groove in the PDMS layer.

(5) Stripping PDMS: and cutting off the PDMS containing the pattern by using a scalpel, completely stripping the PDMS from the silicon wafer, and finally punching the micro-cavity channel by using a puncher to manufacture an inlet and an outlet.

And (3) carrying out plasma treatment on the PDMS cavity channel with the special structure and the glass slide, wherein the power of the plasma treatment is 150W, the duration is 2min, then adhering the PDMS cavity channel end downwards on the glass slide, and baking in an oven at 80 ℃ till the temperature is over night.

Example 2 resonant microbubble array platform set-up

The structure of the resonant microbubble array platform is shown in fig. 3, and the experimental platform comprises the following devices: a signal generator, a power amplifier, a PDMS cavity channel, a micro-injection pump, a pipeline, a cell recovery container and a body wave transducer.

Wherein:

1. the signal generator is used for providing a sine wave signal for the body wave transducer.

2. The power amplifier amplifies the energy of the signal generated by the signal generator.

3. The micro-syringe pump can continuously inject liquid into the PDMS cavity.

And 4, the PDMS cavity channel comprises an array microstructure, when liquid is injected into the PDMS cavity channel, no liquid flows into the microstructure, so that a micro-microbubble is formed, wherein the top view after injection is shown in FIG. 5, and it can be seen that no liquid exists at the microstructure arrays on two sides of the cavity channel, so that the microbubble is formed.

5. The tubing is used to transport the liquid and the solution with the cells.

EP tubes use the liquid after cell lysis back and forth.

7. The bulk wave transducer is used for generating bulk waves, the bulk waves can cause the resonance of micro bubbles generated by the PDMS microstructure, the micro bubble vibration can cause the flow of liquid in the liquid, and the shearing force generated by the flow of the liquid can cause the cell lysis.

Parameter selection and flow field characterization

As shown in fig. 4, due to the interface boundary effect, when liquid is injected into the PDMS cavity, micro-pores located on the sidewall of the PDMS cavity capture micro-bubbles, and the micro-bubbles resonate and generate a flow field when the array micro-bubbles are excited by a single ultrasonic vibration source at a specific frequency. In the experiment, the width and the height of the micro-hole on the side wall of the PDMS channel are 40.8 μm and 50 μm in sequence, the frequency of the bulk wave transducer is 107kHz, and the input effective voltage value is 144 Vpp. PS bead trace particles are added into the PDMS cavity, and the microbubble peripheral flow field is symmetrically and uniformly distributed under the ultrasonic stimulation. And the size of the shear stress in the flow field can be calculated through parameters such as the liquid velocity of the flow field. The shear stress is calculated by the formula

S＝2π^3/2ε²(ρ_ff³μ)^1/2/R_b，

Wherein R is_bIs the radius of the microbubble, f is the resonance frequency of the microbubble, ρ_fWhere the density of the liquid, μ is the relative fluid velocity and ε is the amplitude of the oscillation of the microbubbles.

EXAMPLE 3 lysis of cells

The resonance microbubble array platform is adopted to carry out cell lysis experiments, and the cell lysis efficiency and the integrity of DNA after cell lysis are quantitatively analyzed.

A solution containing cells that simultaneously label both the Calcein-AM and PI fluorescent probes was injected from the inlet using a micro-syringe pump. The cell-containing solution in the lumen was retained in the lumen for 3 minutes. And the lysate was recovered through the outlet and the cracking efficiency was analyzed. Wherein Calcien-AM is a viable cell indicator that freely penetrates the intact cell membrane, is hydrolyzed by intracellular esterase to calcein, and emits green fluorescence. When the cell membrane is damaged, PI can smoothly enter the cell to be combined with DNA or RNA to emit red fluorescence, and the cell lysis efficiency can be calculated by combining a cell sensitive field diagram. As can be seen by calculation, a lysis rate of 99% can be achieved when the cells are retained in the lumen for 3 minutes.

Claims

1. An apparatus for use in cracking, the apparatus comprising;

1) a cavity body of the micro-bubble array is formed, the cavity body is provided with a fluid cavity channel, and two sides of the fluid cavity channel are provided with micro-structure cavity bodies which are communicated with the cavity channel and protrude outwards, and the micro-structure cavity bodies can capture and keep micro-bubbles;

preferably, the width and height of the microstructure cavity are respectively 1 μm to 100 μm, preferably 40 to 50 μm.

2. The device of claim 1, wherein the cavity forming the array of microbubbles has at least one inlet and one outlet; preferably, the inlet is communicated with a liquid inlet device, and the outlet is communicated with a cell lysate recovery device.

3. The apparatus of claim 1, wherein the bulk wave generating device is a bulk wave transducer; preferably, the frequency of the wave transducer is 100-120 kHz.

4. The apparatus of claim 1, further comprising a signal generator and a power amplifier.

5. The apparatus of claim 4, wherein the signal generator is configured to generate a sine wave signal and send the sine wave signal to the power amplifier.

6. The apparatus of claim 5, wherein the power amplifier is configured to amplify the sine wave signal and send the amplified sine wave signal to the bulk wave transducer.

7. The apparatus of claim 1, wherein the cavity forming the array of microbubbles has one or more fluid channels, and the microstructure cavities are staggered on both sides of the channels.

8. A method for lysing cells, viruses, bacteria or tissues, using a device according to any one of claims 1 to 7, comprising the steps of:

1) placing a liquid containing cells, viruses, bacteria or tissues into a lumen of the device, the micro-structured lumen of the lumen capturing and generating microbubbles;

2) generating body waves through a body wave generating device, enabling the microbubbles to resonate, causing the vibration of the solution in the cavity, and generating a flow field in the cavity for cracking cells or viruses;

preferably, during the generation of the body waves by the body wave generating device, a liquid comprising cells, viruses, bacteria or tissues is continuously transported through the lumen.

9. The method of claim 8, further comprising step 3) recovering the lysate.

10. Use of the device according to any one of claims 1-7 for cell lysis, bacterial lysis, viral lysis or tissue understanding.