Gene transfection devices and gene transfection methods

Abstract

The application provides a gene transfection device and a gene transfection method. The gene transfection device comprises an ultrasonic transfection chip for generating ultrasonic waves and a reaction channel in contact with the ultrasonic transfection chip to receive the ultrasonic waves, and the reaction channel is used for loading a cell solution. The device provides a novel, simple, high-throughput, fast, safe and efficient solution for cell transfection, has the characteristics of wide application range, simple operation, good repeatability, strong stability and the like, and can flexibly realize efficient transfection of adherent cells and suspension cells, and has good applicability to various difficult-to-transfect cells such as stem cells, primary cells and tumor cells.

Description

Technical Field

[0001] This application belongs to the field of biomedical ultrasound technology, specifically, it relates to a gene transfection device and a gene transfection method. Background Technology

[0002] Gene transfection refers to the technology of transporting and delivering biologically functional nucleic acids into cells, enabling specific nucleic acids to maintain their biological functions within the cell. Gene transfection is applied in various technical fields, including gene function regulation, gene therapy, vaccine production and drug development, and industrial and agricultural biotechnology. Gene transfection technology has wide applications, especially in gene therapy, playing a crucial role in areas such as the treatment of genetic diseases, tumors, and nervous system diseases, as well as vaccine research and drug screening. Gene transfection can be classified according to physical and chemical transfection methods. Physical transfection methods have significant advantages due to their low toxicity, high efficiency, and good reproducibility. Currently, common physical transfection methods include electroporation, gene gun transfection, and microinjection, which have laid a foundation for physical transfection methods. However, due to difficulties in optimization, such as expensive equipment, high operational difficulty, low throughput, and significant cell damage, physical transfection methods are not widely used in practice. Currently, the most commonly used in vitro gene transfection methods include electroporation, gene gun transfection, microinjection transfection, liposome transfection, and calcium phosphate precipitation transfection.

[0003] Electroporation transfection: This method uses high-voltage electric pulses to create transient, reversible pores in the cell membrane, allowing exogenous nucleic acids to enter the cell. The cell membrane then closes within a certain time, trapping the exogenous nucleic acids and preventing further leakage. The designed plasmid is then expressed through transcription, translation, and protein modification. While widely used, electroporation transfection has limitations: ① High cytotoxicity and poor cell viability: Irreversible cell membrane perforation and oxidative stress caused by the high-voltage electric field result in cell viability of only 30%-70%, especially damaging primary cells and stem cells, where viability can drop below 30%. ② Unstable perforation efficiency: Highly dependent on parameters such as electric field strength and pulse duration, requiring optimization for different cell types. Cell heterogeneity also affects experimental reproducibility. ③ Difficult nucleic acid integration: Low stable transfection efficiency and difficulty in delivering large biological molecules. ④ Demanding operation: Requires optimization of buffer composition, meticulous operation, short window period, and complex handling of adherent cells. ⑤ Cost and throughput limitations: Limited by specialized equipment and disposable consumables, costs are high, and the amount of cells processed per cycle is small, making it unsuitable for large-scale experiments. ⑥ High safety risks: Involves high-voltage operation and the potential for aerosol contamination. These drawbacks limit experiments requiring sensitive cells or high cell viability.

[0004] Gene gun transfection is a physical transfection technique that directly introduces exogenous nucleic acids (DNA, RNA, etc.) into cells or tissues. Its core principle involves using high-speed microparticles (usually gold or tungsten particles) to carry the exogenous nucleic acid. Accelerated by a high-pressure power system, these microparticles penetrate the cell membrane (or tissue surface), delivering the nucleic acid into the target cells, thus achieving transfection. While gene gun transfection is simple to operate and widely applicable, it has significant drawbacks: ① High-speed microparticles penetrating the cell membrane can cause severe mechanical damage to cells, resulting in unpredictable organelle damage and low organelle survival rates (especially in primary cells). ② Transfection efficiency is uneven; the distribution of microparticles in the target tissue and cell population cannot be guaranteed to be completely uniform, leading to localized over-transfection and cell death, while other areas are poorly transfected or even untransfected. This phenomenon is particularly pronounced when processing large-volume samples or complex tissues. ③ The gene gun equipment, gold particles, and high-pressure gas consumables are expensive, making it unsuitable for high-throughput and large-scale research. ④ Exogenous genes not integrated into the genome are mostly expressed transiently, which may lead to false positive results. Subculturing of transfected cells or tissues is still necessary to ensure stable gene expression, consuming significant effort and time. ⑤ Different living tissues exhibit heterogeneity, with significant differences in stiffness, water content, and blood supply. Appropriate parameters need to be selected, and application to living tissues may induce inflammation or structural damage. ⑥ Operational parameters require strict control, resulting in a high technical threshold. These limitations make it more suitable for transient transfection and tissue surface manipulation, but less adaptable to applications requiring high survival rates, stable expression, or large-scale experiments.

[0005] Microinjection transfection is a transfection method that uses micromanipulation techniques to directly inject exogenous nucleic acids (DNA, RNA, etc.) or other biomolecules into a single cell. Its core feature is precise targeting of a single cell, enabling highly efficient delivery to specific cells and playing a crucial role in stem cell research, developmental biology research, and transgenic animal preparation. While microinjection transfection can achieve precise single-cell delivery, it has significant limitations: ① It has an extremely high technical threshold, requiring skilled operators who manually control microneedles under a microscope. Only 50-100 cells can be processed per hour, and cell quality cannot be guaranteed, resulting in low experimental efficiency and failing to meet the needs of large-scale transfection. ② Severe cell damage occurs during microneedle puncture, with cell membrane rupture leading to fluid loss, a major factor affecting cell survival, especially for small cells less than 10 μm in diameter, significantly reducing the success rate. ③ High equipment costs require specialized equipment such as inverted microscopes, micromanipulators, and microneedle pullers, and maintenance costs are also high. ④ The operation is risky; microneedles are prone to clogging or breakage, requiring frequent replacement. Prolonged operation can also lead to visual fatigue and hand tremors, affecting the success rate. ⑤ The application is limited, mainly suitable for larger cells (such as oocytes and fertilized eggs) or specific tissues (such as embryos). It is difficult to efficiently transfect suspended cells or primary cell populations. These drawbacks make it only suitable for special scenarios such as transgenic animal preparation and single-cell functional research, and difficult to extend to routine experiments.

[0006] Liposome transfection is a chemical transfection technique that uses synthetically produced liposomes to deliver exogenous nucleic acids (DNA, RNA, etc.) or proteins into cells. Its core principle is to utilize the similarity between liposomes and cell membranes, introducing the target gene into cells through membrane fusion or endocytosis. It is suitable for gene delivery in in vitro cell culture. Due to its low cost and simple operation, this transfection method is still widely used in gene expression analysis, protein production, and other fields. While liposome transfection is simple to operate, it has significant drawbacks: ① Cationic liposomes easily cause cell membrane damage and oxidative stress, leading to cytotoxicity (especially with significantly lower transfection efficiency for primary cells and suspension cells compared to tumor cells and adherent cells). ② Transfection efficiency is limited by cell type (e.g., almost ineffective for stem cell transfection) and requires strict optimization of the DNA / liposome ratio and avoidance of serum. ③ It is costly.

[0007] Calcium phosphate precipitation transfection: A classic chemical transfection technique that, based on an appropriate pH environment, forms tiny DNA-calcium phosphate co-precipitates. These co-precipitates are adsorbed onto the cell surface via electrostatic attraction and subsequently endocytosed into the cell, thereby achieving the delivery and expression of exogenous genes. Calcium phosphate precipitation transfection is widely used due to its low cost, but it has significant drawbacks: ① Transfection efficiency is generally low (1%-5%), especially for primary and suspension cells, where it is even less effective or ineffective. ② The reproducibility of the experiment is highly dependent on experimental conditions such as pH and calcium ion concentration; even small deviations can lead to unstable results. ③ High concentrations of calcium phosphate precipitation can easily induce cytotoxicity, resulting in poor cell condition or even cell death. ④ The procedure is cumbersome, requiring precise control of buffer preparation, mixing speed and force, as well as incubation time, placing high demands on experimental operation and process.

[0008] Viral transfection: Viral transfection is a gene delivery method that uses modified replication-deficient viruses as vectors to simulate the natural viral infection process, efficiently delivering exogenous genes into eukaryotic cells. Its core principle is to achieve target gene expression through viral binding to cell receptors, invasion, nucleic acid release, and subsequent integration or free existence. This method has significant advantages such as extremely high transfection efficiency, efficient infection of difficult-to-transfect and non-dividing cells, stable or long-term expression, and applicability to various in vivo and in vitro scenarios. However, this viral transfection method also carries risks such as insertional mutations leading to tumorigenesis, strong immunogenicity of some viruses, high biosafety requirements, limited gene packaging capacity, long preparation cycles and high costs, and the potential for gene silencing and cytotoxicity. It is currently a highly efficient but strictly controlled technique in gene research and therapy. Summary of the Invention

[0009] The technical problem addressed by this application is: how to provide a simple, high-throughput, rapid, safe, and efficient gene transfection device and gene transfection method.

[0010] This application provides a gene transfection device, the gene transfection device comprising: An ultrasonic transfection chip, wherein the ultrasonic transfection chip is used to generate ultrasonic waves; A reaction chamber that contacts the ultrasonic transfection chip to receive ultrasonic waves, and the reaction chamber is used to load cell solution.

[0011] Optionally, the ultrasonic transfection chip is a surface acoustic wave chip.

[0012] Optionally, the surface acoustic wave chip includes: piezoelectric substrate; Interdigital transducers are attached to the piezoelectric substrate.

[0013] Optionally, the interdigital transducer is a unidirectional interdigital transducer.

[0014] Optionally, the reaction cavity is placed above the surface acoustic wave chip, and the bottom of the reaction cavity is attached to the interdigital transducer.

[0015] Optionally, there are multiple surface acoustic wave (SAW) chips and reaction cavities, with each reaction cavity placed above the SAW chip in a corresponding manner.

[0016] Optionally, the gene transfection device further includes a PCB substrate, and the ultrasonic transfection chip is disposed on the PCB substrate.

[0017] This application also provides a gene transfection method using a gene transfection device, the gene transfection method comprising: The reaction chamber containing the cell solution is placed in the ultrasonic transfection chip and comes into contact with the ultrasonic transfection chip; The ultrasonic transfection chip is connected to a signal so that it generates ultrasonic waves, which drive the cell solution to perform gene transfection.

[0018] The gene transfection device and gene transfection method provided in this application have the following technical advantages: This device provides a novel, simple, high-throughput, rapid, safe and efficient solution for cell transfection. It features wide applicability, easy operation, good reproducibility and strong stability. It can flexibly achieve efficient transfection of adherent cells and suspension cells, and has good applicability to a variety of difficult-to-transfect cell types such as stem cells, primary cells and tumor cells. Attached Figure Description

[0019] Figure 1 This is a side view of a gene transfection apparatus according to one or more embodiments.

[0020] Figure 2 This is a top view of an ultrasound transfection chip of a gene transfection apparatus according to one or more embodiments.

[0021] Figure 3 This is a flowchart illustrating the preparation of a gene transfection device according to one or more embodiments.

[0022] Figure 4 This is a three-dimensional structural diagram of the reaction cavity of a gene transfection device according to one or more embodiments.

[0023] Figure 5 This is a schematic diagram illustrating the cell transfection effect of a gene transfection device according to one or more embodiments. Detailed Implementation

[0024] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the scope of this application.

[0025] Before describing the various embodiments of this application in detail, the technical concept of this application is first briefly described: Current gene transfection methods either have complex operation, low throughput, slow transfection speed, or high safety risks. Therefore, this application provides a gene transfection device. The key improvement lies in the generation of ultrasound waves by an ultrasonic transfection chip. The ultrasound waves stimulate the cell solution in the reaction cavity, inducing controllable deformation and transient increase in permeability of the cell membrane based on the ultrasonic mechanical effect. This achieves rapid, efficient, and highly stable delivery of exogenous genes, with wide applicability, simple operation, and good reproducibility. The specific principle of the gene transfection device of this application will be described below with reference to more embodiments.

[0026] Specifically, such as Figure 1 , Figure 2 and Figure 3 As shown, the gene transfection device of this embodiment includes an ultrasonic transfection chip 10 and a reaction chamber 20. The ultrasonic transfection chip 10 is used to generate ultrasonic waves, and the reaction chamber 20 is in contact with the ultrasonic transfection chip 10 to receive ultrasonic waves. The reaction chamber 20 is used to load cell solution.

[0027] The principle behind ultrasound-assisted gene transfection in cells is as follows: the mechanical action of ultrasound causes the local tension of the cell membrane to exceed a critical threshold, disrupting the arrangement of lipid molecules and forming transient hydrophilic channels. When the stimulus is removed, the lipid molecules spontaneously rearrange, and the membrane structure recovers its integrity within seconds to minutes, preventing long-term cell damage. The actin cytoskeleton network beneath the cell membrane undergoes transient depolymerization and reconstruction during ultrasound-induced membrane deformation, further reducing membrane mechanical resistance and promoting channel formation and transmembrane transport of exogenous molecules.

[0028] In one or more embodiments, the gene transfection device further includes a PCB substrate, on which an ultrasonic transfection chip is disposed.

[0029] In one or more embodiments, such as Figure 2 As shown, the ultrasonic transfection chip 10 is a surface acoustic wave (SAW) chip. Exemplarily, the SAW chip includes a piezoelectric substrate 11 and an interdigital transducer 12, the interdigital transducer 12 being attached to the piezoelectric substrate 11. The interdigital transducer 12 is a unidirectional interdigital transducer. The interdigital transducer 12 includes a stacked adhesive layer and a conductive layer, the adhesive layer being fixedly connected to the piezoelectric substrate 11.

[0030] For example, the reaction channel 20 is placed above the surface acoustic wave (SAW) chip, and the bottom of the reaction channel 20 is attached to the interdigital transducer 12. In some embodiments, there are multiple SAW chips and reaction channels 20, with each reaction channel 20 placed one-to-one above the SAW chip to accommodate different gene transfection requirements. Different numbers of reaction channels can be freely combined according to experimental needs, enabling flexible selection of multiple channels to meet the needs of small-scale, pilot-scale, and large-scale transfection.

[0031] For example, the ultrasonic wave pattern of the surface acoustic wave chip can be any waveform such as sine wave, square wave, pulse wave, amplitude-modulated wave, etc., all of which can achieve good transduction effect. The piezoelectric substrate 11 can be lithium niobate, quartz, zinc oxide film, alumina film, or PZT piezoelectric ceramic. The adhesion layer can be made of chromium, titanium, molybdenum, tantalum, or their alloys; the conductive layer can be made of metals such as gold, copper, aluminum, or platinum.

[0032] The design and fabrication of the ultrasonic transfection chip 10 are crucial. The structural parameters of the ultrasonic transfection chip 10, such as electrode thickness and linewidth, number of electrode pairs, and acoustic aperture, all have complex effects on acoustic perforation and transfection results. The optimal combination of chip structural parameters needs to be determined through theoretical simulation and extensive testing experiments. The ultrasonic transfection chip 10 needs to achieve impedance matching with the signal source amplifier, thus requiring the design of additional electrode lines. However, the structure and position of these additional lines, in turn, affect the acoustic and temperature field distribution in the chip's working area, thereby impacting cell transfection performance, and further design optimization is required. A single-finger, single-phase mode, consisting of a single interdigital transducer (IDT) and a reflective grating, improves energy conversion efficiency.

[0033] For example, refer to Figure 3 The fabrication process of the ultrasonic transfection chip 10 is described below. The piezoelectric substrate 11 can be a lithium niobate wafer, the adhesion layer of the interdigital transducer is a nickel-chromium alloy, and the conductive layer is a gold electrode. The specific steps are as follows: (1) Cleaning: The lithium niobate wafers were thoroughly cleaned with anhydrous ethanol, dried with nitrogen, and then placed on a 65°C heating plate to dry the residual moisture for later use.

[0034] (2) Coating: Fix the clean lithium niobate wafer onto the spin coater and drop 500 μL of AZ5214 positive photoresist into the center; first spin coat at a low speed of 500 rpm for 15 s, then spin coat at a high speed of 3000 rpm for 30 s, and use a profilometer to detect the photoresist thickness.

[0035] (3) Exposure: The lithography machine is preheated for 30 minutes. The coated lithium niobate wafer is placed on the chuck, covered with a custom film, and exposed at 65 mJ / cm². 2 The dose has been applied and the exposure is complete.

[0036] (4) Post-baking: Place the exposed lithium niobate wafer on a 65°C heating plate and bake for 3-5 minutes.

[0037] (5) Development: Immerse the lithium niobate wafer in AZ300 MIF developer and gently shake until the photoresist in the pattern area is completely removed; after removal, rinse with deionized water to remove residual developer and blow dry with nitrogen.

[0038] (6) Hardening: Place the lithium niobate wafer on a heating plate and continue baking for 3-5 minutes, then store it in a dust-free box for later use.

[0039] (7) Sputtering: Ni-chromium alloy and gold electrode are sputtered sequentially in a magnetron sputtering instrument, with the Ni-chromium alloy target sputtered for 1 min (thickness 50 nm) and the gold target sputtered for 4 min (thickness 200 nm).

[0040] (8) Stripping and cleaning: Immerse the sputtered surface acoustic wave (SAW) chip in acetone solution and ultrasonically clean it in a water bath for 5–7 minutes until all excess metal is completely stripped off. Clean it in sequence with anhydrous ethanol and deionized water, and dry it with nitrogen gas. The entire process is carried out in a fume hood.

[0041] (9) Cutting: The SAW chip is cut along the preset scribing line using a precision dicing machine and then packaged for later use.

[0042] like Figure 4 As shown, in one or more embodiments, the reaction channel 20 generally includes a cylindrical portion 21 and an edge portion 22 located at the upper end of the cylindrical portion 21. The cylindrical portion 21 is a container for loading cell solution, and the edge portion 22 can serve as a handle for easy handling by the operator. The shape, aperture, height, position, and arrangement of the reaction channel 20 are set according to actual needs. The material of the reaction channel 20 can be polydimethylsiloxane (PDMS), polymethyl methacrylate (PMMA), cyclic olefin copolymer (COC), hydrogel, agarose, gelatin, etc., which have good ultrasonic conductivity and thermal stability, and can achieve similar ultrasonic mechanical effects and transfection functions. A molded channel is formed using photolithography, materials are prepared according to the ratio, and then the reaction channel is fabricated through steps such as casting, drying and curing, demolding, and drilling.

[0043] For example, PDMS is used as the material for the reaction channel 20 to illustrate the preparation process of the reaction channel 20. The specific steps are as follows.

[0044] (1) Preparation of PDMS curing adhesive: Mix PDMS prepolymer liquid and curing agent at a mass ratio of 10:1 and stir thoroughly; pour into a clean petri dish, vacuum dryer for 30 min to remove air bubbles; place the petri dish in a horizontal oven and let it stand at room temperature for 1 h to make the adhesive layer uniform; bake at 80℃ for 1.5 h until fully cured.

[0045] (2) Preparation of PDMS reaction channels: Channels of a specified diameter were prepared on a 2mm thick PDMS film using a punch. Surface debris and impurities were removed using 3M tape. Two PDMS films of different thicknesses were placed in an oxygen plasma cleaner for 1 minute, quickly bonded together, and then baked in an 80℃ oven for 10 minutes to achieve a strong bond. The channels were then cut into uniform disposable PDMS reaction channels using a spatula. These channels exhibit high stability, good pressure resistance, and are not prone to leakage.

[0046] (3) Pretreatment of PDMS reaction cavity cell culture: The cavity was cleaned with 75% ethanol and pure water, dried at 80℃ and then transferred to a clean bench for ultraviolet sterilization for 30 min. Poly-L-lysine solution was added and incubated in a CO2 incubator for 2 h to enhance the hydrophilicity of the cavity and the cell adhesion ability.

[0047] In one or more embodiments, after preparing the ultrasound transfection chip 10 and the reaction channel 20, the reaction channel 20, which is filled with cell solution, is placed on and in contact with the ultrasound transfection chip 10, so that the bottom of the reaction channel 20 is completely attached to the interdigital transducer 12 of the ultrasound transfection chip 10. Next, the ultrasound transfection chip is connected to a signal generator to generate ultrasound waves, which drive the cell solution to perform gene transfection.

[0048] When conducting ultrasound experiments, it is necessary to optimize the ultrasound experimental parameters, including ultrasound voltage, excitation time, intracavitary solution volume, and cell type. Appropriate ultrasound parameters should be selected according to requirements for cell transfection. For example, the calcein-AM / PI double staining method is used to characterize cell membrane perforation, where calcein-AM (green fluorescence) marks cell viability and PI (red fluorescence) marks changes in membrane permeability. Red-green fluorescence co-localization is used as the criterion for determining reversible cell perforation. Ultrasound parameters (including electrical power, pulse frequency, and exposure time) with the highest cell viability and optimal perforation efficiency are selected as the transfection parameters.

[0049] Finally, the cell transfection effect was evaluated. After transfection, the PDMS cavity was removed and placed in a culture dish, which was then transferred to a cell culture incubator for further culture. The transfection efficiency and viability of the cells were observed using a fluorescence microscope at 24h, 48h, and 72h, and flow cytometry could be used to achieve precise quantitative assessment of cell viability and transfection efficiency.

[0050] For example, such as Figure 5As shown, (a) is a bright-field image of the cells, (b) is a cell transfection with a plasmid tagged with green fluorescent GFP (GFP-DNA), and (c) is a superposition of the bright-field and fluorescence images, intended to observe the transfection efficiency of the cells. The PCDNA plasmid was successfully transfected into MCF-7 cells and expressed efficiently using an ultrasound system. The fluorescence field image shows that after ultrasound, the perforation efficiency of MCF-7 cells was approximately 90%, and the PCDNA plasmid was successfully and efficiently expressed, thus verifying that the device can achieve efficient, rapid, and safe gene transfection.

[0051] Compared with existing technologies such as electroporation, gene gun, microinjection, liposomes, calcium phosphate precipitation, and viral transfection, the gene transfection device and gene transfection method of this embodiment have significant advantages: (1) Minimal cell damage and high survival rate: The gentle ultrasonic mechanical effect is used to achieve reversible perforation, avoiding severe cell damage caused by high voltage electric field, mechanical puncture and chemical reagents. It is especially suitable for fragile cells such as stem cells and primary cells.

[0052] (2) High transfection efficiency and strong universality: It is not limited by cell type, cell state, or adherence / suspension, and can still achieve high-efficiency transfection of difficult-to-transfect cells.

[0053] (3) No chemical toxicity, no immunogenicity, and no risk of insertion mutation: Pure physical transfection, without the use of viral vectors and chemical transfection reagents, is safer and suitable for gene therapy and clinical translation.

[0054] (4) Flexible throughput and scalable: It supports free combination of 4–120 channels, which can be used for both micro-cell screening and large-scale preparation, solving the problems of low throughput and difficulty in scaling up traditional methods.

[0055] (5) Simple operation, short time consumption, and good repeatability: no complicated pretreatment is required, and transfection can be completed in a few minutes; the device standardization is high, and the experimental stability and repeatability are significantly better than traditional methods.

[0056] (6) Low equipment cost, easy integration and easy promotion: The standard photolithography mold process is used to prepare reaction cavity structures with different arrangements, sizes, shapes and positions, realizing a low-cost, high-throughput and consistent cell transfection cavity preparation method. No expensive special equipment is required. The system has a simple structure and high integration, and can be widely used in ordinary laboratories and industrialization scenarios.

[0057] (7) Strong functional scalability: In addition to gene transfection, it can also be used for various scenarios such as cell washing, cell sorting, cell perforation, cell delivery, cell imaging, and 3D cell culture, and has the value of a multi-purpose platform.

[0058] The specific embodiments of this application have been described in detail above. Although some embodiments have been shown and described, those skilled in the art should understand that modifications and improvements can be made to these embodiments without departing from the principles and spirit of this application as defined by the claims and their equivalents, and such modifications and improvements should also be within the protection scope of this application.

Claims

1. A gene transfection device, characterized in that, The gene transfection device includes: An ultrasonic transfection chip, wherein the ultrasonic transfection chip is used to generate ultrasonic waves; A reaction chamber that contacts the ultrasonic transfection chip to receive ultrasonic waves and is used to load cell solution.

2. The gene transfection device according to claim 1, characterized in that, The ultrasonic transfection chip is a surface acoustic wave chip.

3. The gene transfection device according to claim 2, characterized in that, The surface acoustic wave chip includes: piezoelectric substrate; Interdigital transducers are attached to the piezoelectric substrate.

4. The gene transfection device according to claim 3, characterized in that, The interdigital transducer is a unidirectional interdigital transducer.

5. The gene transfection device according to any one of claims 2 to 4, characterized in that, The reaction cavity is placed above the surface acoustic wave chip, and the bottom of the reaction cavity is attached to the interdigital transducer.

6. The gene transfection device according to claim 5, characterized in that, The number of surface acoustic wave chips and reaction cavities is multiple, and each reaction cavity is placed on top of the surface acoustic wave chip in a one-to-one correspondence.

7. The gene transfection device according to claim 1, characterized in that, The gene transfection device also includes a PCB substrate, and the ultrasonic transfection chip is disposed on the PCB substrate.

8. A gene transfection method using a gene transfection device according to any one of claims 1 to 7, characterized in that, The gene transfection method includes: The reaction chamber containing the cell solution is placed in the ultrasonic transfection chip and comes into contact with the ultrasonic transfection chip; The ultrasonic transfection chip is connected to a signal so that it generates ultrasonic waves, which drive the cell solution to perform gene transfection.

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