Processing device and method based on a specific ultrasound device

Abstract

The application relates to a method for processing cells based on a special ultrasonic device, comprising the following steps: placing the special ultrasonic device and target cells in a positionally fixed state, and at least placing the special ultrasonic device in a liquid environment; driving the special ultrasonic device to generate an acoustic beam flow to act on a target position of the target cells, so as to change the state of the target cells, including: acting on a first position of the surface of the target cells, so that the first position of the surface of the target cells is impacted by the pulsed acoustic beam flow, to generate one of the following changes: the surface of the target cells is deformed, a gap is generated at the first position, the first position is cut, and a corresponding state responding to the stimulation is entered; wherein the corresponding state responding to the stimulation comprises one of the following: activating a mechanical gate channel of the cells, recombining a cell skeleton, activating a signal transduction pathway, and changing a secretion activity. The application can realize high focusing processing of the cells, and has strong controllability.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of micro-electro-mechanical system, in particular to a processing device and method based on a special ultrasonic device. BACKGROUND

[0002] With the development of micro-electro-mechanical system, cell analysis or processing platforms based on micro-electro-mechanical system are produced, such as analysis platforms based on microfluidic chips combined with lasers, analysis platforms combined with probes, etc.

[0003] The platform based on micro-electro-mechanical system also includes an acoustic-based platform, which has the advantages of low power consumption, small damage to cells, and strong versatility. However, due to the divergent characteristics of sound propagation, the jet flow generated by acoustic radiation force is usually in a cluster state, and the focusing is insufficient, which leads to the difficulty of the existing acoustic-based cell analysis or processing platform in realizing the operation of high focusing on single cell level, and the controllability is high, or it needs to be realized by other components or structures, which is inconvenient to implement.

[0004] Therefore, how to provide an acoustic-based solution that can achieve high focusing on single cell level, and is convenient to implement and has strong controllability, is a technical problem to be solved. SUMMARY

[0005] In view of the above problems of the prior art, the present application provides a processing device and method based on a special ultrasonic device, which can achieve high focusing on single cell level, and is convenient to implement and has strong controllability.

[0006] The first aspect of the present application provides a method for processing cells based on a special ultrasonic device, comprising: making the special ultrasonic device and the target cell in a position-fixed state, and at least making the special ultrasonic device in a liquid environment, and making the target cell in the action range of the acoustic beam flow of the special ultrasonic device; driving the special ultrasonic device to generate an acoustic beam flow in the liquid environment, the acoustic beam flow acting on the target position of the target cell to change the state of the target cell.

[0007] From the above, through the acoustic beam flow with high focusing, high processing precision can be achieved, such as single cell level precision, and the controllability of the acoustic beam flow is strong, which is convenient to implement.

[0008] As a possible implementation manner of the first aspect, the driving the special ultrasonic device to generate the acoustic beam flow in the liquid environment, the acoustic beam flow acting on the target position of the target cell to change the state of the target cell comprises: periodically driving the special ultrasonic device to generate a pulsed acoustic beam flow in the liquid environment, the acoustic beam flow acting on a first position of the surface of the target cell, the first position of the surface of the target cell being impacted by the pulsed acoustic beam flow to generate at least one of the following changes: the surface of the target cell is deformed, the surface of the target cell is perforated, a hole is generated at the first position, the first position is cut, and the target cell enters a corresponding state in response to the stimulation; wherein the target cell entering the corresponding state in response to the stimulation comprises at least one of the following: activating a mechanical gate channel of the cell, recombining a cytoskeleton of the cell, activating a signal transduction pathway, and changing a secretion activity.

[0009] As a possible implementation manner of the first aspect, the driving the special ultrasonic device to generate the acoustic beam flow in the liquid environment, the acoustic beam flow acting on the target position of the target cell to change the state of the target cell comprises: periodically driving the special ultrasonic device to generate a pulsed acoustic beam flow in the liquid environment, the acoustic beam flow acting on a first position of the surface of the target cell, the first position of the surface of the target cell being impacted by the pulsed acoustic beam flow to generate at least one of the following changes: the surface of the target cell is deformed, the surface of the target cell is perforated, a hole is generated at the first position, the first position is cut, and the target cell enters a corresponding state in response to the stimulation; wherein the target cell entering the corresponding state in response to the stimulation comprises at least one of the following: activating a mechanical gate channel of the cell, recombining a cytoskeleton of the cell, activating a signal transduction pathway, and changing a secretion activity.

[0010] As a possible implementation manner of the first aspect, the first position of the surface of the target cell comprises a position where a part of the surface of the target cell is perpendicular to the acoustic beam flow.

[0011] As a possible implementation manner of the first aspect, the driving the special ultrasonic device to generate the acoustic beam flow in the liquid environment, the acoustic beam flow acting on the target position of the target cell to change the state of the target cell comprises: periodically driving the special ultrasonic device to generate a pulsed acoustic beam flow in the liquid environment, the acoustic beam flow acting on a first position of the surface of the target cell, the first position of the surface of the target cell being impacted by the pulsed acoustic beam flow to generate at least one of the following changes: the surface of the target cell is deformed, the surface of the target cell is perforated, a hole is generated at the first position, the first position is cut, and the target cell enters a corresponding state in response to the stimulation; wherein the target cell entering the corresponding state in response to the stimulation comprises at least one of the following: activating a mechanical gate channel of the cell, recombining a cytoskeleton of the cell, activating a signal transduction pathway, and changing a secretion activity.

[0012] As a possible implementation manner of the first aspect, the driving the special ultrasonic device to generate the acoustic beam flow in the liquid environment, the acoustic beam flow acting on the target position of the target cell to change the state of the target cell comprises: periodically driving the special ultrasonic device to generate a pulsed acoustic beam flow in the liquid environment, the acoustic beam flow acting on a first position of the surface of the target cell, the first position of the surface of the target cell being impacted by the pulsed acoustic beam flow to generate at least one of the following changes: the surface of the target cell is deformed, the surface of the target cell is perforated, a hole is generated at the first position, the first position is cut, and the target cell enters a corresponding state in response to the stimulation; wherein the target cell entering the corresponding state in response to the stimulation comprises at least one of the following: activating a mechanical gate channel of the cell, recombining a cytoskeleton of the cell, activating a signal transduction pathway, and changing a secretion activity.

[0013] As a possible implementation manner of the first aspect, the target cell is located on a binding object, and the binding object has a part of the target cell in the liquid environment, or the target cell is located on a binding object, and the binding object has a part of the target cell not in the liquid environment, and the binding object has a part of the target cell in a position that can be impacted by the acoustic beam flow to be out of the liquid environment.

[0014] As a possible implementation manner of the first aspect, the driving the special ultrasonic device to generate the acoustic beam flow in the liquid environment comprises: the special ultrasonic device is driven with a first power cycle, in particular, the special ultrasonic device is driven to work in at least one driving cycle, each driving cycle comprises a first stage and a second stage, in the first stage, the special ultrasonic device is driven to work to generate special ultrasonic waves of 0.5-30 GHz, in the second stage, the driving of the special ultrasonic device is stopped.

[0015] The first power has a size that makes the special ultrasonic device generate the acoustic beam flow with a desired intensity in the first stage; the duration of the first stage is less than a threshold or the duty cycle of the first stage in a driving cycle is less than a threshold, so as to suppress the special ultrasonic device from generating secondary flow in the liquid environment or / and suppress the heat generated by the special ultrasonic device.

[0016] The second aspect of the present application provides a special ultrasonic device-based processing device, comprising: a stage, on which a target cell in a liquid environment can be placed; a mechanical arm, a probe is mounted on the movable end of the mechanical arm, and a special ultrasonic device is mounted on the end of the probe; the special ultrasonic device at the end of the probe is moved above the stage and extended into the liquid environment by the mechanical arm, so that the target cell is in the action range of the acoustic beam flow of the special ultrasonic device; the special ultrasonic device is driven to be driven to generate an acoustic beam flow in the liquid environment, the acoustic beam flow acts on a target position of the target cell to change the state of the target cell.

[0017] As a possible implementation manner of the second aspect, the stage is made of transparent material; an image acquisition device is arranged below the stage, the image acquisition device comprises at least one lens, and the at least one lens can be adjusted to face the stage.

[0018] From the above, the acoustic beam flow with high focusing property acts on the target position of the target cell to change the state of the target cell, which can achieve single-cell level precision, and the controllability (such as intensity, diameter, frequency, etc.) of the acoustic beam flow is strong, which can simulate the actions of beating (or knocking, beating), poking, pricking, etc. by pulse impact, the target cell changes state directly under the action of the acoustic beam flow or in response to the stimulation of the impact, or the acoustic beam flow acts on the cell at a certain angle to simulate the effect of shoveling cells and separate the cells from the attached objects. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 are images and schematic diagrams of the jet phenomenon and the secondary flow phenomenon;

[0020] Figure 2 is a diagram of a sound beam flow generated by an embodiment of the present application in a liquid environment;

[0021] Figure 3 is a timing schematic diagram of an output signal of a driving device of a special ultrasonic device provided by an embodiment of the present application;

[0022] Figure 4 is a schematic diagram of a driving device of a special ultrasonic device provided by an embodiment of the present application;

[0023] Figure 5 is a schematic diagram of a processing device based on a special ultrasonic device provided by a second embodiment of the present application;

[0024] Figure 6 is a partial enlarged schematic diagram of Figure 5

[0025] Figure 7 is a schematic diagram of a probe of a special ultrasonic device with different characteristics that can be replaced in the present application;

[0026] Figure 8 is a diagram of a sound beam flow generated in a fourth embodiment of the present application;

[0027] Figure 9 is a diagram of different action areas under different intensities in a fourth embodiment of the present application;

[0028] Figure 10 is a parameter schematic diagram corresponding to each diagram; Figure 9

[0029] Figure 11 is a diagram of areas acted on under different proportions in a first stage of a driving cycle;

[0030] Figure 12 is a diagram of a relationship between driving power and sound beam flow speed or force under different distances;

[0031] Figure 13 is a size schematic diagram of a special ultrasonic device;

[0032] Figure 14 is a diagram of an embodiment acting on a single cell.

[0033] The reference signs are as follows: 1- stage, 11- electric control box, 2- mechanical arm, 21- mechanical arm controller, 22- rotating part, 23- probe, 24- probe end, 3- image acquisition device, 31- lens.

[0034] ​​It should be understood that the size and shape of each block in the structural schematic diagram above are only for reference, and should not constitute an exclusive interpretation of the embodiments of the present application. The relative position and inclusion relationship between the blocks presented in the structural schematic diagram are only used to represent the structural association between the blocks, and not to limit the physical connection mode of the embodiments of the present application. DETAILED DESCRIPTION

[0035] The technical solutions provided by the present application will be further described below in combination with the drawings and examples. It should be understood that the system structure and business scenarios provided in the embodiments of the present application are mainly used to illustrate possible implementation modes of the technical solutions of the present application, and should not be interpreted as the only limitation of the technical solutions of the present application. Those skilled in the art can know that with the evolution of system structure and the appearance of new business scenarios, the technical solutions provided by the present application are also applicable to similar technical problems.

[0036] It should be understood that the technical solutions provided in the embodiments of the present application include the method and device for processing cells based on the special ultrasonic device. Since the principles of solving problems of these technical solutions are the same or similar, in the introduction of the following specific embodiments, some repeated parts may not be described again, but should be regarded as mutual reference and mutual combination between these specific embodiments.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present application belongs. If there is any inconsistency, the meaning explained in the specification or the meaning derived from the content described in the specification shall prevail. In addition, the terms used herein are only for the purpose of describing the embodiments of the present application, and are not intended to limit the present application. In order to accurately describe the technical content in the present application, and to accurately understand the present application, before the specific embodiments are described, the terms used in the specification are first explained as follows:

[0038] 1) Special ultrasonic device: a high-frequency resonator, which can be a device based on the piezoelectric effect, generating mechanical vibration by applying voltage. In this application, a piezoelectric resonator is used, which generates special ultrasonic waves of not less than 0.5 gigahertz (GHz) during operation. Preferably, a piezoelectric resonator is used, which generates special ultrasonic waves of not less than 1 GHz and not more than 30 GHz during operation, for example, 2G-2.5 GHz. Such piezoelectric resonators can be, for example, Surface Acoustic Wave (SAW) devices, Bulk Acoustic Wave (BAW) devices, etc. For example, when BAW, it can be a Film Bulk Acoustic Resonator (FBAR), a Solidly Mounted Resonator (SMR), or a Lamb Wave Resonator (LWR). For the sake of convenience, the piezoelectric resonator that can generate special ultrasonic waves of not less than 0.5 GHz is referred to as a special ultrasonic device in the following.

[0039] 2) Jet phenomenon: a phenomenon that occurs when the acoustic waves of the special ultrasonic device act on the liquid. The area vibration generated by the working interface of the special ultrasonic device can form a traveling wave in the liquid, and continuously push the local liquid in the liquid environment (i.e. volume force), so that at least part of the local liquid moves linearly along the direction of the acoustic wave propagation. This linear motion phenomenon is called jet phenomenon.

[0040] Secondary flow phenomenon: including vortex and thermal backflow, is another phenomenon that occurs when the special ultrasonic device acts on the liquid, including vortex (or micro eddy) caused by local circulation driven by the jet, and thermal backflow caused by heat generated by the special ultrasonic device.

[0041] The jet phenomenon and the secondary flow phenomenon can be seen in the images and schematic diagrams shown in Figure 1 The schematic diagram of the use of vortex to capture microparticles shown in Figure 1 The schematic diagram of the use of vortex to capture microparticles shown in

[0042] 3) Acoustic beam flow: in this application, the acoustic beam flow is generated based on the special ultrasonic device. In this application, the acoustic beam flow is a unique type of jet phenomenon, characterized by high-speed movement of the fluid along the direction of acoustic wave propagation and fine cylindrical shape of the jet in its travel. The acoustic beam flow is in a substantially laminar state with the surrounding liquid before it is significantly attenuated, and has low mixing degree with the surrounding liquid. As shown in Figure 2 The image of the generated acoustic beam flow taken by a high-speed camera. Figure 2 As can be seen, the acoustic beam flow has focusing property and is no longer like the cluster of jet in Figure 1 In Figure 2 The secondary flow phenomenon in Figure 1 cannot be seen obviously.

[0043] The generating condition of the acoustic beam stream of the embodiments of the present application is: driving the special ultrasonic device to work in at least one driving period with the first power, each driving period including a first stage and a second stage (see the driving unit output signal part in Figure 3 ), driving the special ultrasonic device to work to generate a special ultrasonic liquid environment of 0.5-30 GHz in the first stage, and stopping driving the special ultrasonic device in the second stage; wherein the first power is large enough to make the special ultrasonic device generate a focused columnar acoustic beam stream of the vertical solid-liquid interface in the first stage. Wherein the duration of the first stage under the first power is less than a threshold or the duty cycle of the first stage in a driving period is less than a threshold, so as to suppress the temperature of the special ultrasonic device within a controllable temperature threshold, and to suppress the special ultrasonic device from generating secondary flow (including vortex flow and backflow) in the liquid environment.

[0044] Wherein the intensity of the acoustic beam stream is related to the duration of the first stage (or the duty cycle in a driving period) and the intensity of the first power, and the duration of the first stage and the first power constitute factors of how much energy is input into the special ultrasonic device in a driving period. For example, the shorter the first stage duration, the lower the temperature and the secondary flow can be suppressed, but the acoustic beam stream intensity is also lower (the first stage duration is short, which means the cumulative energy time is short), and the higher the first power, the higher the acoustic beam stream intensity, and the higher the temperature of the special ultrasonic device. Therefore, by applying a higher first power and a shorter first stage, a higher intensity acoustic beam stream can be obtained, and the temperature and the secondary flow can be effectively suppressed. Therefore, when it is necessary to control the acoustic beam stream intensity in real time, the preferred solution is to use a shorter first stage with a fixed value (such as the duty cycle of the first stage value in the period is a fixed value), and control the intensity of the generated acoustic beam stream by applying different first power. The second preferred solution is to apply the same first power to adjust the different first stage values (such as the duty cycle of the first stage value in the period) to control the intensity of the generated acoustic beam stream, or the third preferred solution is to adjust the first stage value and the first power at the same time to control the intensity of the generated acoustic beam stream.

[0045] When the liquid environment is determined and the special ultrasonic device is selected, the relationship between the acoustic beam stream intensity and the control parameters (the first power and / or the period signal composed of the first stage and the second stage) can be calibrated. In one case, if the period signal is a fixed value (i.e. the first stage and the second stage are fixed values), the first power and the acoustic beam stream intensity can be calibrated to obtain the first power size and the acoustic beam stream intensity.

[0046] 4) about the first stage in the driving cycle: in the first stage of the driving cycle, the driving unit continuously outputs several signals. For example, when the signal generator outputs a 1GHz signal, assuming the duration of the first stage is 1 microsecond, the driving unit will output 1000 signals in the first stage time, and the power loaded in the first stage time will be loaded on the 1000 signals output to drive the special ultrasonic device. In the second stage of the driving cycle (corresponding Figure 3 to the low level of the switch signal), no driving signal is output.

[0047] For specific cases, for example, only the first stage is executed once, which is equivalent to or considered in this application as only the driving cycle is executed once, and such cases are also within the protection scope of this application.

[0048] 5) driving device of the special ultrasonic device: as Figure 4 shown, it includes a control unit and a driving unit, and the driving unit includes a signal generator and a power amplifier. Its signal output principle can be referred to Figure 3 shown.

[0049] The signal generator is used to generate high-frequency signals, and the frequency of the original high-frequency signals generated is the same as or close to the operating frequency (or the inherent frequency) of the special ultrasonic device as the load. Among them, the waveform of the signal generator can be a square wave (such as Figure 3 shown), a sawtooth wave, a sharp pulse (or a triangular wave), a ladder wave, a positive cosine wave, or a half wave, etc. The output signal of the corresponding driving unit is modulated into a sawtooth wave, a sharp pulse (or a triangular wave), a ladder wave, a positive cosine wave, or a half wave, etc.

[0050] The power amplifier is used to amplify the signal to be output, so as to drive the special ultrasonic device.

[0051] The control unit can be a switching power supply, which can control the output switch signal. The switch signal can be a periodic signal, and the switch-on stage (such as the conduction stage of the switching power supply transistor) corresponds to the high level, and the first stage in Figure 3 corresponds to the low level, and the second stage in Figure 3 corresponds to the low level. The control unit can also control the operating voltage or amplification factor input to the power amplifier to realize the output of different powers. The output power is the first power mentioned above. The first power can also be understood as the average energy density input to the special ultrasonic device in a driving cycle (a driving cycle is composed of a first stage and a second stage). Since the size of the energy is related to the power and the time, the amount of energy input to the special ultrasonic device in a driving cycle is related to the first power, the duration of the first stage, or the duty cycle thereof.

[0052] 6) Impact resistance: In this application, it is used to reflect the stability and resistance of the target object under the action of a sound beam flow with a set intensity. The value of resistance can be times, duration, etc.

[0053] For example, when the sound beam flow is intermittent (i.e. pulsed, such as a first stage proportion of 50%, 20%, etc.), the target object is a cell, and when the cell membrane is perforated after being impacted by the pulsed sound beam flow n times, it can be called that the cell membrane resistance under the corresponding sound beam flow parameters (sound beam flow parameters include at least one of the following: power, driving period, first stage proportion, etc., and the distance and angle directly opposite to the target object) is n, which specifically reflects the resistance of the cell membrane to maintain structural integrity under the action of the sound beam flow parameters. For another example, when the sound beam flow is continuous (the first stage proportion is 100%, which is a special case), the resistance value can also be the duration.

[0054] For another example, in addition to the resistance of the above reaction to maintain structural integrity (integrity), since the target object can also be other particulate matter, etc., it can also be the resistance reflecting the stability of the target object in a fixed position or posture under the action of the above sound beam flow parameters, the resistance of the target object to maintain the shape under the action of the sound beam flow parameters, etc.

[0055] 7) Corresponding state of cell response to stimulation: When cells are subjected to continuous mechanical stimulation, they will respond to these stimuli through specific mechanisms, which is also called mechanical signal transduction. For example, the following A-E are several possible response modes of cells:

[0056] A, activation of mechanosensitive channels: There is a special protein on the cell membrane--mechanosensitive channels, which can directly sense and respond to changes in mechanical force. When the cell membrane is subjected to extrusion, stretching or other forms of mechanical stimulation, these channels can open or close, allowing ions such as sodium (Na+), calcium (Ca2+) or chlorine (Cl-) to enter or exit the cell, changing the potential of the cell membrane, triggering electrical signal transduction, and further affecting the physiological activities within the cell.

[0057] B, reorganization of cytoskeleton: Cytoskeleton (including microfilaments, microtubules and intermediate filaments) is crucial for maintaining cell morphology and transmitting mechanical force. Mechanical stimulation can promote the reorganization of cytoskeleton, such as promoting the polymerization and depolymerization of actin, affecting the adhesion, migration and division of cells, etc.

[0058] C. Activation of signaling pathways: Mechanical stimuli can also activate various intracellular signaling pathways, such as those regulated by Rho family GTPases, affecting gene expression, cell proliferation, differentiation, and apoptosis, etc. For example, osteoblasts, upon sensing mechanical forces, can promote bone formation and repair through these pathways.

[0059] D. Changes in secretory activity: Certain cells can change their secretion patterns upon mechanical stimulation, releasing growth factors, cytokines, and other bioactive molecules, which can influence the behavior of surrounding cells and tissue repair and remodeling.

[0060] E. Apoptosis or damage: If mechanical stimulation is too strong or lasts too long, it can cause excessive cellular stress, potentially leading to apoptosis or damage.

[0061] Among them, the above-mentioned sound beam flow and the related technology of driving the special ultrasonic device to generate the sound beam flow can also be referred to the related introduction in Chinese patent application No. CN2024108322079.

[0062] The scheme for processing cells based on the special ultrasonic device provided in the present application can control the special ultrasonic device to generate a high-concentration sound beam flow in a liquid, so as to process target cells through the sound beam flow to change the state of the target cells. The sound beam flow described in the present application has a small action area (or a small diameter or cross section), a high pressure, and a fast response (i.e., can be generated instantaneously and disappear instantaneously, or a pulsed sound beam flow with a high frequency, which can be on the order of microseconds). Among them, the pulsed sound beam flow generated by periodically driving the special ultrasonic device acts on the target cells to simulate the actions of tapping (or knocking, beating), poking, and pricking, etc. on the target cells through the pulsed impact, and the target cells change the state directly under the action of the sound beam flow or in response to the stimulation of the impact. The scheme based on the high-concentration sound beam flow in the present application can realize the way of high-frequency impact acting on the target cells, and the response speed of the special ultrasonic device generating the sound beam flow is fast.

[0063] In the following, the scheme provided in the present application will be described in detail with reference to the accompanying drawings and examples.

[0064] The first embodiment of the present application provides a method for processing cells based on a special ultrasonic device, comprising:

[0065] The special ultrasonic device and the target cells are in a positionally fixed state, and at least the special ultrasonic device is in a liquid environment, and the target cells are in the action range of the sound beam flow of the special ultrasonic device;

[0066] The special ultrasonic device is driven to generate a sound beam flow in the liquid environment, and the sound beam flow acts on the target position of the target cells to change the state of the target cells.

[0067] In some embodiments, the ultrasound device can be driven in a periodic manner to generate a pulsed acoustic beam in the liquid environment, the pulsed acoustic beam acting on a first location on the surface of the target cell to cause the first location on the surface of the target cell to be impacted by the pulsed acoustic beam to cause at least one of the following: a deformation of the surface of the target cell, a pore to be created on the surface of the target cell, a pore to be created at the first location, a cut to be made at the first location, and a corresponding state of the target cell in response to the impact.

[0068] In some embodiments, the corresponding state of the target cell in response to the impact includes at least one of the following: a mechanical gate of the cell to be activated, a cytoskeleton of the cell to be reorganized, a signal transduction pathway of the cell to be activated, and a secretion activity of the cell to be changed.

[0069] In some embodiments, the target cell is much larger than a cross-sectional area of the acoustic beam, and the first location is on the surface of the target cell. In this case, the impact of the pulsed acoustic beam on the target cell can be modeled as a poke or a stab. In other embodiments, a ratio of a surface area of the target cell to the cross-sectional area of the acoustic beam is less than one (e.g., 50%), and the surface of the target cell is completely covered by the cross-sectional area of the acoustic beam. In this case, the impact of the pulsed acoustic beam on the target cell can be modeled as a whole knock or a hit.

[0070] In some embodiments, when the cross-sectional area of the acoustic beam is small or relatively small compared to the surface area of the target cell, a stronger acoustic beam is more likely to cause a pore or a cut on the surface of the target cell. In some embodiments, when the cross-sectional area of the acoustic beam is large or relatively large compared to the surface area of the target cell, or when a weaker acoustic beam is used, the target cell is more likely to enter a corresponding state in response to the impact and activate a mechanical gate of the cell. Of course, both situations can exist at the same time.

[0071] In some embodiments, the target cell can be a single cell as described above. In other embodiments, the cross-sectional area of the acoustic beam generated can be controlled to cover two or more single cells, and the target cell is the two or more cells.

[0072] In some embodiments, the impact on the target cell can also be measured to determine a number of impacts required to cause a certain state change.

[0073] Wherein, for each change produced by the target cell as listed above, further experiments can be conducted to test which change is more prominent when different parameters of the acoustic beam are applied, and the trend of the change of the acoustic beam and each change. The parameters of the acoustic beam applied herein can include at least one of the following: driving power of the special ultrasound device, driving period, time length or duty cycle of the first and second stages of the driving period, action time length, distance and / or angle between the special ultrasound device and the target position, etc.

[0074] In some embodiments, a substance in the liquid environment can also be introduced into the target cell through the pore or the mechanical gate channel, or a substance in the liquid environment can be introduced into the target cell through the transmission of the acoustic beam. In some embodiments, the substance introduced into the target cell through the transmission of the acoustic beam is accelerated by the acoustic beam. In some embodiments, when the substance is accelerated by the acoustic beam and introduced into the target cell, it can be directly perforated into the cell membrane.

[0075] In some embodiments, the substance in the liquid environment can be, for example, ions (such as Na+, Ca2+, Cl-, etc.), genetic material (such as DNA or RNA), drugs, particles (such as vesicles or particle-shaped drugs), etc., to achieve the effect of physiological activity of the target cell after being introduced into the target cell, transfection of the target cell, drug injection, etc. In some embodiments, the pore or the mechanical gate channel of the cell can be self-closed after the impact of the acoustic beam is ended.

[0076] In some embodiments, the first position of the surface of the target cell includes the position of the part of the surface of the target cell perpendicular to the acoustic beam, i.e., the position directly opposite to the acoustic beam, to achieve a better impact of the acoustic beam. In some embodiments, in addition to achieving the rapid impact of the perpendicular position (i.e., the front position), the rapid impact of the side position can also be included, and continuous impact can achieve dynamic extrusion of the cell, continuous mechanical stimulation, etc.

[0077] In some embodiments, the special ultrasound device can be periodically driven to generate a pulsed acoustic beam in the liquid environment, and the acoustic beam acts on the second position of the surface of the target cell, which is the binding position of the target cell and the binding object, so that the second position is impacted by the pulsed acoustic beam and is separated from the binding object.

[0078] In some embodiments, further comprising collecting the target cells detached from the binding object. For example, a pipette can be arranged in the vicinity of the target cells, preferably in the direction of movement of the target cells after detachment, such as on the other side of the second position of the target cells.

[0079] In some embodiments, the binding object can be a carrier such as a glass slide, and the target cells adhered to the carrier can be detached by the acoustic streaming. In other embodiments, the binding object can be another cell to which the target cell is bound, and the target cell can be detached from the other cell by the acoustic streaming.

[0080] In some embodiments, when the target cell is detached from another cell, one specific principle can be that the cells are bound by multiple protein contact interfaces (i.e., multivalent interaction binding), and the acoustic streaming can be used to rapidly cut the second protein under the condition that the first protein is cut and has not recovered, and so on to cut the bound proteins to achieve efficient cell peeling.

[0081] In some embodiments, the target cells are on a binding object, and the binding object has part of the target cells in the liquid environment. For example, a carrier with target cells is placed in a vessel with a liquid environment, and the ultrasonic device is inserted into the liquid environment to act on the target cells on the carrier.

[0082] In some embodiments, the target cells are on a binding object, and the binding object has part of the target cells not in the liquid environment, and the binding object has part of the target cells in a position that can be reached by the acoustic streaming out of the liquid environment. For example, a carrier with target cells is placed upside down (with the target cells facing down) above a vessel with a liquid environment, and is not immersed in the liquid environment, and the ultrasonic device is inserted into the liquid environment to generate acoustic streaming upward and breaking through the surface of the liquid environment to act on the target cells on the carrier. Another example is that the ultrasonic device is configured with a liquid supply device, and the liquid supply device provides a liquid environment when the ultrasonic device is working (such as the ultrasonic device is located in an open microcavity, and the microcavity is connected to the liquid supply device), and the acoustic streaming generated by the ultrasonic device can break through the surface of the liquid environment to act on the target cells on the carrier.

[0083] In some embodiments, the device for implementing the above-mentioned method for processing cells based on the special ultrasonic device can be a mechanical arm as described later. In some embodiments, it can also be handheld, for example, the configuration described above is provided with a liquid environment, the special ultrasonic device is located in an open microcavity, the microcavity is connected to the liquid supply device to provide a liquid environment when the special ultrasonic device is working, and the microcavity can be located at the end of a handheld part, and a driving circuit and the like can be arranged in the handheld part. In some embodiments, it can also be implemented in a microfluidic system, for example, located at the bottom of a flow channel or a microcavity, and the microfluid channel can flow through a liquid containing target cells.

[0084] The second embodiment of the present application provides a processing device based on a special ultrasonic device, which can be referred to as shown in Figure 5 、 Figure 6 The processing device can include:

[0085] A carrier 1, on which a target object in a liquid environment can be placed, the target object including target cells;

[0086] A mechanical arm 2, a movable end of the mechanical arm 2 is equipped with a probe 23, and a probe end 24 is equipped with a special ultrasonic device, and the probe 23 can be moved or stopped above the carrier by the mechanical arm 2;

[0087] The special ultrasonic device is used to generate a sound beam flow in the liquid environment to act on the target object.

[0088] In some embodiments, the probe end 24 is equipped with one or more special ultrasonic devices. In some embodiments, when there are multiple special ultrasonic devices, an array arrangement can be formed, wherein the working surfaces of at least two special ultrasonic devices are arranged in the same plane, or the working surfaces of at least two special ultrasonic devices are arranged at an acute angle. In some embodiments, the multiple special ultrasonic devices can have the same or different parameters to generate sound beams of the same or different intensities and diameters under the same driving parameters.

[0089] In some embodiments, the special ultrasonic device equipped on the probe end 24 can be located on both sides of the probe end, so that the probe 23 can be clamped and fixed at the rotating part 22 after rotating 180 degrees along its axis, that is, the quick selection of the working special ultrasonic device can be realized.

[0090] In some embodiments, when the probe has multiple special ultrasonic devices as described above, the working of each special ultrasonic device can be driven respectively, and the special ultrasonic device to be used can be moved to the desired position and attitude by the movement of the mechanical arm to act on the target object.

[0091] In some embodiments, the stage 1 is made of transparent material; and an image acquisition device 3 is arranged below the stage 1, the image acquisition device 3 comprises at least one lens 31, which is adjustable to face the stage 1. In this way, the process and result of the sound beam flow acting on the target object can be acquired by the image acquisition device 3 below to be used for analysis or control of the working state of the special ultrasonic device and the position of the moving end of the mechanical arm based on the obtained image data or analyzed data.

[0092] In some embodiments, the image acquisition device 3 can be a microscope with multiple lenses 1, which are assembled on a rotating part to realize switching of the working lens and make the working lens face upward.

[0093] In some embodiments, the image acquisition device 3 can also be arranged beside or above (e.g. obliquely above) the stage, and the lens 31 of the image acquisition device 3 is adjustable to face the target object on the stage 1 to acquire images. In some embodiments, the image acquisition device 3 can also comprise multiple lenses 31 arranged at different positions (e.g. below, beside or obliquely above the stage).

[0094] In some embodiments, the mechanical arm 2 comprises a three-axis mechanical arm arranged beside the stage 1, and the movable end of the three-axis mechanical arm is equipped with a rotating part 22, and the rotating part 22 is equipped with the probe 23. In some embodiments, the rotating part 22 can rotate along a horizontal axis. In some embodiments, the rotating part 22 has a clamping part to facilitate replacement of the probe 23. In some embodiments, the probe end 24 as shown in the right part of FIG. 8, wherein the probe 23 can be composed of two detachable parts, one part is used to be assembled on the rotating part 22, and the other part has the probe end 24 and is equipped with the special ultrasonic device, so that the probe 23 or the part with the probe end 24 can be replaced as needed. Figure 7 In some embodiments, the probe end 24 as shown in the right part of FIG. 8, wherein the probe 23 can be composed of two detachable parts, one part is used to be assembled on the rotating part 22, and the other part has the probe end 24 and is equipped with the special ultrasonic device, so that the probe 23 or the part with the probe end 24 can be replaced as needed. Figure 7 In some embodiments, the probe end 24 as shown in the right part of FIG. 8, wherein the probe 23 can be composed of two detachable parts, one part is used to be assembled on the rotating part 22, and the other part has the probe end 24 and is equipped with the special ultrasonic device, so that the probe 23 or the part with the probe end 24 can be replaced as needed.

[0095] In some embodiments, the stage 1 can move horizontally; wherein the stage 1 can be assembled on a horizontal guide rail, so that it can be driven by a motor to move along the guide rail to adjust the target object on the stage to a desired position, and can be equipped with an electric control box 11, which can have a control part to control the movement of the stage 1.

[0096] In some embodiments, the movement and rotation of the probe 23 can be achieved by the three-axis mechanical arm and the rotating part 22. In some embodiments, the three-axis mechanical arm is an x, y, z axis mechanical arm, which achieves the translation of the probe 23, and the rotating part 22 can be the rotating part along the horizontal axis as described above, which achieves the rotation of the probe 23, so as to adjust the special ultrasonic device at the end of the probe 24 to a desired position. The desired position can make the special ultrasonic device contact or immerse in the liquid environment as described above. And the mechanical arm controller 21 can be provided to control the posture of the mechanical arm.

[0097] In some embodiments, the special ultrasonic device is driven in a first power cycle to generate the acoustic beam flow. For details, refer to the description of the acoustic beam flow, driving cycle and driving device above. In addition, it should be noted that in subsequent experiments, the driving cycle also uses the first stage with a proportion of 100% (which can be understood as continuous driving without forming a pulse mode) under the allowable condition of the temperature of the special ultrasonic device.

[0098] In some embodiments, the liquid environment, for example, the liquid contained in a container placed on the object table 1, and the target object is located in the container. Wherein, the container is also made of transparent material, so as to facilitate the image acquisition of the image acquisition device 3 below the object table 1. In other embodiments, the liquid environment refers to the liquid on the surface of the target object. The liquid can be the liquid carried by the surface of the target object, or the liquid continuously sprayed or drained to the target position of the target object when the special ultrasonic device is working.

[0099] In some embodiments, the target object is fixed relative to the probe. For example, the target object can be attached to the surface of a device (such as a glass slide) or the surface of other tissues (such as organic tissues), and the device or organic tissues can be fixedly placed in the container as described above.

[0100] In some embodiments, the direction of the generated acoustic beam flow can be perpendicular to the surface of the special ultrasonic device. For example, when the special ultrasonic device is a bulk acoustic wave device (BAW), the piezoelectric constant d33 of the piezoelectric layer (such as AlN layer) of the BAW has a direction characteristic along the thickness direction, that is, the vibration direction of the BAW piezoelectric layer, and the acoustic wave transmission direction is perpendicular to the BAW. In other embodiments, the direction of the acoustic beam flow can form an acute angle with the surface of the special ultrasonic device. For example, when the special ultrasonic device is a surface acoustic wave device (SAW), the angle between the acoustic beam flow direction, that is, the acoustic wave transmission direction, and the surface of the SAW is about 30 to 50 degrees.

[0101] In some embodiments, the special ultrasonic device can be a polygon, especially a polygon with an odd number of sides. For example, in one embodiment, it can be a regular pentagon, or an irregular pentagon. In other embodiments, it can be an olive shape, a triangle, an ellipse, a rhombus, a semicircle, or any polygon of any size in any shape combination.

[0102] wherein the area (herein refers to the radial dimension, non-thickness dimension, i.e. the device interface dimension) of the special ultrasonic device is about 10-1000000 μm 2 , preferably about 100-40000 μm 2 , more preferably 1000-10000 μm 2 . Wherein, in order to intuitively understand the size of the special ultrasonic device, Figure 12 the size diagram of the special ultrasonic device as a one-dimensional acoustic wave resonator is shown. Wherein, the size of the special ultrasonic device is negatively correlated with the achievable resonant frequency, so that reducing the size can increase the resonant frequency. In other embodiments, by changing the shape of the device, such as the number of sides of a polygon, the angle between adjacent sides, and the length of each side, the shape formed can also be used to change the resonant frequency of the special ultrasonic device.

[0103] Wherein, the diameter of the acoustic beam generated is related to the parameters of the special ultrasonic device itself, the smaller the area (refers to the radial dimension, or the interface dimension) of the special ultrasonic device, the smaller the diameter of the acoustic beam. Smaller microscale also has better acoustic focusing effect. Wherein, under the same first power application, the area of the special ultrasonic device adopts smaller microscale, has stronger acoustic beam, the diameter of the acoustic beam is also smaller, and the response speed (the time of generating the acoustic beam) is also faster.

[0104] In some embodiments, the intensity (or force, speed) of the acoustic beam acting on the target object at a certain position in the liquid is related to the following factors: the liquid properties (such as viscosity) of the liquid environment, the distance from the target object or the target position, the angle of the acoustic beam and the target object or the target position, the controllable parameters for generating the acoustic beam (the first power for driving the special ultrasonic device, or the periodic signal composed of the first stage and the second stage in the driving cycle), the properties of the special ultrasonic device itself (size, resonant frequency), etc. Therefore, when designing the relative position of the special ultrasonic device and the target object, the non-variable factors (such as liquid properties, properties of the special ultrasonic device itself) can be determined first, and then the variable factors (i.e. factors used as control parameters in the specific control process, such as distance, angle, or controllable parameters for generating the acoustic beam) are calibrated (calibration can be done through simulation, experiment or calculation) to determine the force on the target object or the target position under different values of the variable factors, and then based on this, how to control the relative distance, angle or controllable parameters for generating the acoustic beam between the special ultrasonic device and the target position is designed to controllably generate acoustic beams with desired intensity and direction (angle / position) acting on the target object or the target position.

[0105] The third embodiment of the present application provides a method for processing cells based on a special ultrasonic device, using any of the above-mentioned processing devices based on a special ultrasonic device or its optional embodiments, the processing method comprising the following steps:

[0106] S1: placing a target object in a liquid environment on a stage, the target object comprising target cells.

[0107] S2: controlling a mechanical arm to adjust the pose of the special ultrasonic device assembled at the end of the probe, the pose being such that the special ultrasonic device is angled towards the target object and at least a part of the special ultrasonic device is located in the liquid environment.

[0108] S3: causing the special ultrasonic device to generate a sound beam flow in the liquid environment to act on the target object.

[0109] In some embodiments, the method is used for at least one of the following:

[0110] a. Measuring a certain property parameter of the target object by setting the parameter of the sound beam flow to continuously or intermittently impact the target object.

[0111] b. Causing the target object to detach from the surface of another object to which it is attached by causing the sound beam flow to act on the target position or the target object. In some embodiments, the other object can be a glass slide as described above, or an organic tissue.

[0112] c. Causing the target object to generate a hole at the position of the sound beam flow by causing the sound beam flow to act on the target object.

[0113] d. Causing particles in the liquid environment to enter the target object by causing the sound beam flow to act on the target object.

[0114] e. Applying mechanical stimulation to analyze the target object by causing the sound beam flow to act on the target object, the mechanical stimulation comprising at least one of the following: poking the target object, bombarding the target object, shoveling the target object, and cutting the target object.

[0115] In some embodiments, when used for measuring a certain property parameter of the target object as described in a, the property comprises the impact resistance of the target object under a sound beam flow of a set parameter, and the method comprises:

[0116] S1a: placing an article containing the target object, such as a glass slide with adherent cells (the cells being the target objects), in a container with a liquid.

[0117] S2a: adjusting the positions of the mechanical arm and the stage so that the special ultrasonic device at the end of the probe is immersed in the liquid in the container and is directed towards the target object at a set distance from the target object, and controlling the generation of a sound beam flow of a set parameter to continuously or intermittently impact the target object.

[0118] S3a: continuously capture images by the image capturing device below the stage, and analyze the images to determine the time duration or the number of intervals for the target object to produce a state change, which is used as a property parameter for measuring the impact resistance of the target object under the sound beam flow of the set parameter.

[0119] The state change includes at least one of the following: a change in the integrity of the target object, a change in the position or posture of the target object, and a change in the shape of the target object.

[0120] In some embodiments, when the target object is detached from the surface of another object in b above, the method includes:

[0121] S1b: Place an article containing a target object, such as a glass slide with adherent cells (the cells are the target objects), in a container with a liquid.

[0122] S2b: Adjust the position of the mechanical arm and the stage so that the tip of the probe with the ultrasonic device extends into the liquid in the container and is directed towards the target object at a set distance, and controls the sound beam flow to produce a set parameter to impact the target position continuously or at fixed intervals to act on the target object. The target position can be the position that causes the target object to detach from the surface of another object.

[0123] S3b: Continuously capture images by the image capturing device below the stage, and analyze the images to determine whether the target object is detached from another object (such as a glass slide). Based on the images, at least one parameter in S2b can be automatically or manually adjusted to adjust the impact process or the range of the action area.

[0124] In other embodiments, different target objects have different binding forces (or adhesion forces) with another object, which can be a glass slide or an organic tissue (such as a cultured cell wall), etc. By controlling at least one parameter in S2b to a different value, the desired target object can be selectively detached from a plurality of target objects. For example, a glass slide has target objects with different binding forces. By controlling one or more parameters in S2b, when the sound beam flow reaches a certain value, the target objects with binding forces less than or equal to the value are detached from the glass slide.

[0125] In some embodiments, different target objects can be modified with different amounts / types of antibodies, and the different amounts / types of antibodies can be achieved by the antibodies having different binding forces with another object, which can be achieved by modifying the antibodies with a certain substance, including different substances modified with different amounts / types of antibodies in a liquid, forming different target objects, and then binding the antibodies modified substances to the surface of the target object (i.e. the above-mentioned another object, such as a glass slide) to form a coated substance, wherein the substance is bound to the surface of the target object by the modified antibodies, and the different amounts / types of antibodies modified on the substance exhibit different binding forces of the substance with the target object (i.e. different target objects have different binding forces with another object).

[0126] In some embodiments, when applied to the above-mentioned b, the target object is separated from the surface of the coated another object, and a suction inlet is also provided towards the target object or target position, which is in communication with a negative pressure device, and the peeled target object is collected through the suction inlet. The suction inlet can be in the extension direction of the target object or target position after the acoustic beam flows on the target object or target position. In some embodiments, the suction inlet is located near the position of the captured substance of the second special ultrasonic device described below and is arranged towards this position. In some embodiments, the suction inlet is movable, for example, a suction inlet of a hose which can be moved to the corresponding position by hand.

[0127] In some embodiments, a filter screen is also provided at the suction inlet, and the mesh size of the filter screen matches the size of the target object. In some embodiments, the filter screen is replaceable and can be replaced with a filter screen of a desired mesh size.

[0128] In some embodiments, a channel in communication with the suction inlet is also included, and the channel is provided with at least one of the following:

[0129] 1) A queue device for arranging the flowing target objects in a queue, which includes a sheath flow structure or another special ultrasonic device including a tip (such as a blade or a shuttle). For example, the sheath flow structure can be a sheath flow introduced from the side of the channel, can be a three-dimensional structure such as a sheath flow structure surrounding the channel, or can be a two-dimensional structure such as a sheath flow structure located on both sides of the channel or on one side of the channel. The tip of the special ultrasonic device is located on the downstream side, and the target object is captured by the edge of the tip and released through the tip.

[0130] 2) A detection device for detecting the flowing target objects. It can be a laser detection device, an image detection device, a fluorescence detection device, an electrical detection device, a magnetic detection device (such as used when the substance is combined with a magnetic bead), etc.

[0131] 3) An image acquisition device for acquiring images of the flowing target objects. It can be used for image analysis of the target objects, and can be combined with AI technology such as neural network technology for image analysis.

[0132] In some embodiments, a second special ultrasound device is also included to capture the target object after the peeling; the working surface of the second special ultrasound device is also in the liquid environment, and forms a vortex effect to capture the target object in the liquid environment after the peeling.

[0133] In some embodiments, for the captured target object, other operations can also be performed, for example, when the target object is a cell, at least one of the following can be achieved in cooperation with other microfluidic instruments: cell transfection, cell picking, cell measurement, cell interaction with other substances, cell assembly, cell fusion, cell analysis (gene analysis, protein analysis, cell omics analysis).

[0134] In other embodiments, the binding force between the target object and another object can also be measured and calibrated based on the principles of the above S1b-S3b steps.

[0135] In some embodiments, when the particles carried by the acoustic beam flow in the liquid environment are used to enter the target object in the above d, the method comprises:

[0136] S1d: Place an article containing a target object, such as a glass slide with adherent cells (the cells are target objects), in a container with liquid.

[0137] S2d: Adjust the position of the mechanical arm and the stage so that the special ultrasound device at one end of the probe extends into the liquid in the container and is directed towards the target object at a set distance from the target object. Control the acoustic beam flow with set parameters to continuously or fixedly intermittently impact the target object.

[0138] And the route of the acoustic beam flow in the liquid environment, for example, through a microfluidic channel, a conduit, inject the particles into the route of the acoustic beam flow, or near the special ultrasound device, to be acquired by the acoustic beam flow.

[0139] Based on the acoustic beam flow, the particles carried by the acoustic beam flow are impacted into the target object.

[0140] S3d: Continuously capture images by the image acquisition device below the stage, and analyze whether the particles enter the target object based on the images. Wherein, at least one parameter in S2d can also be automatically or manually adjusted based on image analysis to adjust the impact process.

[0141] In some embodiments, for each of the above embodiments, the acoustic beam flow is controlled by at least one of the following: the driving power of the acoustic beam flow, the number of driving cycles of the acoustic beam flow, the proportion of the first stage in the driving cycle of the acoustic beam flow; wherein the acoustic beam flow acting on the area of the target object is positively correlated with at least one of the following: the driving power of the acoustic beam flow, the continuous number of driving cycles of the acoustic beam flow, the proportion of the first stage in the driving cycle of the acoustic beam flow.

[0142] In order to better understand the present application, the present application also provides a fourth embodiment of the present application, which is the detachment of adherent cells by the generated acoustic beam flow, wherein reference can be made to Figure 8 - Figure 12 . Among them, Figure 8 shows the relative position of the special ultrasonic device and the target object, the generated acoustic beam flow and the target position, and the Figure 8 embodiment uses the acoustic beam flow to detach adherent cells (adherent cells are target objects) from the glass slide, so Figure 8 the acoustic beam flow in Figure 8 the glass slide with the target object is placed horizontally, and the direction of the acoustic beam flow is inclined. In order to compare the pulsed acoustic beam flow with the continuous acoustic beam flow (i.e. the first stage proportion is 100%, that is, the device is always on), first use the continuous acoustic beam flow to act on the adherent cells, and obtain the action range, then use the pulsed acoustic beam flow, and compare and illustrate that reducing the first stage proportion can further have a high-concentration action range.

[0143] Figure 9 shows a diagram of the different sizes of the action area of the acoustic beam flow of different intensities generated by the special ultrasonic device under different first power driving in the always-on state (i.e. the first stage proportion in the driving cycle is 100%), and Figure 9 the cross in each figure shows the widest and longest part of the action area, Figure 10 is Figure 9 the parameter list of the corresponding figure. As can be seen, the smaller the first power, the smaller the acoustic beam flow action area, that is, the more focused. From Figure 10 the always-on acoustic beam flow, it can be inferred that the trend relationship (here referring to positive correlation or negative correlation) between the parameters of the pulsed acoustic beam flow is similar.

[0144] Figure 11 shows a diagram of the action area under different proportions of the first stage in the driving cycle, the first power is 2W, the left side figure the first stage proportion (100us / 1000us)%, the right side figure the first stage proportion 100% (i.e. always on), it can be seen that the left side figure the acoustic beam flow generated by the pulsed driving special ultrasonic device has a smaller action range Figure 11The size of the area (shown by the circle in the image) is significantly reduced due to the relative tilt of the ultrasonic device and the stage (see reference...). Figure 8 (as shown in the positional relationship), therefore, Figure 11 The area indicated by the circle in the image (acquired by the image acquisition device) is offset from the location of the ultrasonic device. Among them, Figure 11 The area size of the Zhongte ultrasonic device is 2k square micrometers, see [reference needed]. Figure 13 A schematic diagram of the dimensions of an ultrasonic device is shown to facilitate understanding of the size of the ultrasonic device of this application. Furthermore, experiments have shown that by reducing the proportion of the first stage and increasing the first power, a high-intensity but relatively small-range acoustic beam can still be obtained.

[0145] Figure 12 This is a graph showing the relationship between the driving power of a special ultrasonic device and the velocity or force of the acoustic beam at different distances between the device and the target object. Figure 12 The 400µm and 1000µm values ​​represent the distance between the ultrasonic device and the target object. It can be seen that the force or velocity of the acoustic beam reaching the target object decreases with increasing distance.

[0146] Figure 14 An image showing the experimental results of applying the method described in this application to single cells is presented. Specifically, the image is a frame from a video taken using a downward-facing microscope. The experimental environment was as follows: 2 mL of PBS buffer and 10 μL of PI (propidium iodide) reagent were added to a 3.5 cm diameter culture dish. The cells used were MCF-7 human breast cancer cells (seeded at a density of 10⁴ and cultured in a dedicated cell culture incubator for 48 h). Before the experiment, the cells were stained with calcein (a cell viability assay reagent) in the dark for 30 min to make them appear green (indicating viability). During the experiment, the culture dish was placed in the processing device based on a supersonic device (e.g., [example device]) provided in the embodiments of this application. Figure 5 On the stage of the device shown, the ultrasonic device of the processing device is adjusted to be vertically downward and directly facing the target cell. The ultrasonic device is adjusted to be 500 micrometers away from the target cell. The driving parameters of the ultrasonic device are: power 1W, signal period 200ms, high-level percentage (i.e., first stage percentage) 25%. The ultrasonic device-based processing device provided in this embodiment can achieve automatic control after inputting parameters. Specifically, it automatically focuses the microscope onto the target object and automatically controls the movement of the robotic arm and / or stage to adjust the attitude and position of the ultrasonic device to be directly facing the target cell at a distance of 500 micrometers.

[0147] Once the ultrasonic device is in place, the above driving parameters drive the device to generate an acoustic beam, which is then captured by an image acquisition device located below the stage for observation. The obtained... Figure 14In the image (i.e. the image of the experimental result) in the figure, the green color is a small tissue on the slide (or indicated by A in the figure), and the green color indicates that the small tissue remains active after the experimental treatment. The red color shows a single cell (or indicated by B in the figure) on the green tissue, which is the target cell directly opposite the sound beam flow, and the single cell turns red due to the introduction of the drug during the experiment. Among them, the observation is carried out by the collected video, specifically: about 2 seconds after the sound beam flow is generated from the special ultrasonic device, the single cell at the target position turns red, indicating that the drug contained in the liquid in the vessel is introduced into the single cell.

[0148] The possible way of introducing the drug into the single cell is as follows:

[0149] Firstly, it is speculated that since the drug is introduced into the single cell 2 seconds after observation (the color of the single cell changes 2 seconds later), it is speculated that the intensity of the sound beam flow controlled in this experiment does not directly penetrate the cell membrane in a single pulse. The intensity makes the single cell be impacted by high frequency, and the effect of the high frequency impact is closer to the pulsed beating (or pounding), so that the cell is stimulated to open the mechanical channel (at least one mechanical channel), and the drug in the liquid environment enters the cell through the mechanical channel (exchange between inside and outside the cell).

[0150] Secondly, in addition to the opening of the mechanical channel, it is also possible that the impact of the sound beam flow provides power for the drug in the liquid to pass through the mechanical channel and enter the cell.

[0151] Thirdly, it is possible that after the cell membrane is loosened by high frequency impact, the drug is directly injected into the cell through the loosened cell membrane by the sound beam flow. For example, in one case, the granular drug in the liquid environment is accelerated by the sound beam flow and injected into the cell. Among them, the granular drug can be located near the surface of the special ultrasonic device, or guided to the surface of the special ultrasonic device through the pipeline, so that when the special ultrasonic device works, the granular drug flows to the root of the sound beam flow and is carried and accelerated by the sound beam flow to inject into the cell.

[0152] The experimental result reflects the processing method based on the special ultrasonic device provided in the embodiments of the present application, and the action precision (which can be understood as the diameter or cross section of the sound beam flow) can reach a single cell. Among them, the intensity (pressure) of the action on the single cell is related to the distance between the special ultrasonic device and the target single cell, the intensity of the sound beam flow generated by the special ultrasonic device, the action mode (intermittent or continuous) of the sound beam flow, and the action time.

[0153] In several embodiments provided in the present application, it should be understood that the disclosed apparatus and method are not limited to the above embodiments, and can also be implemented in other manners. For example, the above-described apparatus embodiments are merely schematic. For example, the division of the units is only a logical function division, and there can be another division manner in actual implementation. For example, a plurality of units or components can be combined or integrated into another system, or some features can be ignored or not executed. In addition, the displayed or discussed coupling or direct coupling or communication connection between the units can be indirect coupling or communication connection through some interfaces, devices or units, and can be electrical, mechanical or in other forms.

[0154] The units described as separate components may or can not be physically separate, and the components shown as units may or can not be physical units, i.e., they can be located in one place or distributed on a plurality of network units. Some or all of the units can be selected according to actual needs to achieve the purpose of the embodiments.

[0155] In addition, the words "first", "second", "third" or "module A", "module B", "module C" and the like similar terms in the specification and claims are only used to distinguish similar objects, and do not represent a specific order or sequence of the objects. It can be understood that the specific order or sequence can be interchanged as permitted, so that the application described herein can be implemented in an order other than that illustrated or described herein.

[0156] In the above description, the labels indicating the steps, such as S10, S20, and the like, do not necessarily mean that the steps are necessarily performed in this order, and the order of the steps can be interchanged or performed simultaneously as permitted.

[0157] The term "comprising" used in the specification and claims should not be interpreted as limited to the listed elements; it does not exclude other elements or steps. It should therefore be interpreted as specifying the presence of the stated features, integers, steps or components as referred to, but does not preclude the presence or addition of one or more other features, integers, steps or components thereof. Thus, the expression "a device comprising means A and B" should not be construed to mean that the device only consists of the means A and B.

[0158] The term "one embodiment" or "an embodiment" in the specification means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the application. Therefore, the appearance of the phrase "in one embodiment" or "in an embodiment" in various places in the specification does not necessarily all refer to the same embodiment, but can refer to the same embodiment. In addition, in one or more embodiments, each particular feature, structure or characteristic can be combined in any appropriate manner as apparent to those skilled in the art from the disclosure.

[0159] It is to be understood that the above description is merely a preferred embodiment of the application and the applied technical principles. Those skilled in the art will understand that the application is not limited to the specific embodiments described herein, and that various obvious changes, modifications and substitutions can be made without departing from the scope of the application. Therefore, although the application has been described in detail through the above embodiments, the application is not limited to the above embodiments, and more other equivalent embodiments can be included without departing from the concept of the application, and all belong to the protection scope of the application.

Claims

1. A method for processing cells based on a special ultrasound device, characterized in that, include: The ultrasonic device and the target cell are in a relatively fixed position, and the ultrasonic device is at least in a liquid environment, and the target cell is within the effective range of the ultrasonic device's acoustic beam. Driving the ultrasonic device to generate an acoustic beam in the liquid environment includes: driving the ultrasonic device to operate at a first power for at least one driving cycle, each driving cycle including a first phase and a second phase, wherein in the first phase the ultrasonic device is driven to generate ultrasonic waves of 0.5-30 GHz acting on the liquid environment, and in the second phase the driving of the ultrasonic device is stopped; wherein the magnitude of the first power is sufficient to enable the ultrasonic device to generate a focused columnar acoustic beam perpendicular to the solid-liquid interface in the first phase; The acoustic beam acts on the target location of the target cell to change the state of the target cell.

2. The method according to claim 1, characterized in that, The method of driving the ultrasonic device to generate an acoustic beam in the liquid environment, the acoustic beam acting on the target location of the target cell to change the state of the target cell, includes: The ultrasonic device is periodically driven to generate a pulsed acoustic beam in the liquid environment. The acoustic beam acts on a first position on the surface of the target cell, causing the first position on the surface of the target cell to be impacted by the pulsed acoustic beam, resulting in at least one of the following changes: deformation of the target cell surface, formation of pores on the target cell surface, formation of pores at the first position, being cut at the first position, or entering a corresponding state in response to the stimulus. Among them, the target cell entering the corresponding state in response to the stimulus includes at least one of the following: activating the cell's mechanogate channels, reorganizing the cytoskeleton, activating signal transduction pathways, and altering secretory activity.

3. The method according to claim 2, characterized in that, It also includes at least one of the following: A substance in the liquid environment is allowed to enter the target cell through the pores or the mechanical gate channel; A substance in the liquid environment is transmitted through the acoustic beam, through the pores or the mechanical gate channel, and enters the target cell.

4. The method according to claim 2, characterized in that, The first position on the surface of the target cell includes the location of the portion of the target cell surface that is perpendicular to the acoustic beam.

5. The method according to claim 1, characterized in that, The method of driving the ultrasonic device to generate an acoustic beam in the liquid environment, the acoustic beam acting on the target location of the target cell to change the state of the target cell, includes: The ultrasonic device is periodically driven to generate a pulsed acoustic beam in the liquid environment. The acoustic beam acts on a second position on the surface of the target cell, which is located at the junction of the target cell and the object, so that the second position is impacted by the pulsed acoustic beam and detached from the object.

6. The method according to claim 5, characterized in that, Also includes: The target cells that have detached from the binding object are collected.

7. The method according to any one of claims 1-6, characterized in that, The target cell is located on the binding object, and the portion of the binding object containing the target cell is in the liquid environment, or... The target cell is located on the bonding object, the portion of the bonding object containing the target cell is not in the liquid environment, and the portion of the bonding object containing the target cell is in a position that can be reached by the acoustic beam exiting the liquid environment.

8. The method according to claim 1, characterized in that, The magnitude of the first power causes the ultrasonic device to generate the acoustic beam of the desired intensity during the first phase; the duration of the first phase is less than a threshold or the duty cycle of the first phase within a drive cycle is less than a threshold, in order to suppress secondary flow generated by the ultrasonic device in the liquid environment and / or suppress the heat generated by the ultrasonic device.

9. A processing device based on a supersonic device, characterized in that, include: A stage on which target cells in a liquid environment can be placed; A robotic arm, the movable end of which is equipped with a probe, the probe tip of which is equipped with a special ultrasound device, the robotic arm allows the special ultrasound device at the probe tip to move above the stage and extend into the liquid environment, so that the target cell is within the effective range of the sound beam of the special ultrasound device; The ultrasonic device is driven to generate an acoustic beam in the liquid environment, the acoustic beam acting on a target location of the target cell to change the state of the target cell; wherein the ultrasonic device is driven to operate at a first power for at least one driving cycle, each driving cycle including a first stage and a second stage, wherein the ultrasonic device is driven to operate in the first stage to generate ultrasonic waves of 0.5-30 GHz acting on the liquid environment, and the driving of the ultrasonic device is stopped in the second stage; wherein the magnitude of the first power is sufficient to enable the ultrasonic device to generate a focused columnar acoustic beam perpendicular to the solid-liquid interface in the first stage.

10. The apparatus according to claim 9, characterized in that, The stage is made of transparent material; An image acquisition device is provided below the stage, and the image acquisition device includes at least one lens, which can be adjusted to face the stage.

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