Affinity analysis method and application

Abstract

The invention relates to an affinity analysis method which comprises the following steps: a carrier attached with a first substance is placed in a liquid environment, part of the first substance is combined with a second substance through affinity, and part of the first substance is combined with a third substance through affinity; the special ultrasonic device is placed in a liquid environment, and the relative position of the special ultrasonic device and the carrier can be adjusted; driving the special ultrasonic device to generate a pulse type sound beam in the liquid environment, and continuously adjusting the relative position of the special ultrasonic device and the carrier, so that the pulse type sound beam acts on the combination position of each first substance and the second substance or the third substance combined with the first substance; and acquiring a continuous image comprising the first substance, and counting the quantity of the second substance and the third substance respectively separated from the first substance under the set quantity of pulses, so as to compare the affinity of the second substance and the third substance with the first substance. According to the analysis method, damage to cells is small, and affinity analysis is accurate.

Description

Technical Field

[0001] This application relates to the fields of microelectromechanical systems (MEMS), acoustic fluid dynamics, cell biology, and molecular biology, and in particular to an affinity analysis method, a method for separating a second substance bound to a first substance, a method for screening a target second substance, and a cell processing device. Background Technology

[0002] In the fields of cell biology and molecular biology, affinity generally describes the attractive force or binding ability between different molecules or structures. Affinity can occur between a variety of different objects, such as cell-cell affinity, cell-protein affinity (e.g., the binding force between cell surface receptors and ligands), protein-protein affinity, protein-nucleic acid affinity (e.g., DNA or RNA), and cell or protein affinity with small molecules.

[0003] When the affinity between proteins can be quantitatively and qualitatively determined, it can be applied to dissociate the desired portion based on the measured affinity (such as dissociating the target protein bound to the cell), or to analyze which protein has a stronger or weaker affinity for the cell, facilitating the selection of such proteins.

[0004] How to provide a cell-minimally damaging and relatively accurate affinity analysis method is the technical problem to be solved in this application. Summary of the Invention

[0005] In view of the above-mentioned problems of the prior art, this application provides an affinity analysis method and application. The solution provided by this application has minimal cell damage and the affinity analysis is more accurate.

[0006] The first aspect of this application provides a method for analyzing affinity, including:

[0007] A carrier containing a first substance is placed in a liquid environment, wherein a portion of the first substance is bound to a second substance by affinity, and a portion of the first substance is bound to a third substance by affinity.

[0008] The ultrasonic device is placed in a liquid environment, and the relative position of the ultrasonic device and the carrier is adjustable;

[0009] The ultrasonic device is driven to generate a pulsed sound beam in the liquid environment, and the relative position of the ultrasonic device and the carrier is continuously adjusted so that the pulsed sound beam acts on the bonding position of each first substance with the second or third substance it is bonded to.

[0010] Acquire a continuous image including the first substance, and count the amount of the second and third substances detaching from the first substance under a set number of pulses, to compare the affinity of the second and third substances for the first substance.

[0011] The second aspect of this application provides a method for analyzing affinity, including:

[0012] A carrier with a first substance attached is placed in a liquid environment, wherein a second substance is bound to the first substance by affinity.

[0013] The ultrasonic device is placed in a liquid environment, and the pulsed sound beam of the ultrasonic device can act on the junction of a first substance and a second substance.

[0014] The driving ultrasonic device generates a pulsed acoustic beam at the junction of the first and second substances in the liquid environment;

[0015] Acquire a continuous image including the first substance, determine the number of pulses of the pulsed acoustic beam when the second substance detaches from the first substance, and the number of pulses is used to characterize the affinity between the first substance and the second substance.

[0016] A third aspect of this application provides a method for analyzing affinity, including:

[0017] A carrier with a first substance attached is placed in a liquid environment, wherein a second substance is bound to the first substance by affinity.

[0018] The ultrasonic device is placed in a liquid environment, and the relative position of the ultrasonic device and the carrier is adjustable;

[0019] The ultrasonic device is driven to generate a pulsed acoustic beam in the liquid environment, and the relative position of the ultrasonic device and the carrier is continuously adjusted so that the pulsed acoustic beam acts on the bonding position of each first substance and the second substance it is bonded to.

[0020] Acquire a continuous image including the first substance, and count the number of times the second substance detaches from the first substance under a set number of pulses. The value of the set number of pulses divided by the number of times the second substance detaches from the first substance is used to characterize the affinity between the second substance and the first substance.

[0021] As one possible implementation of the first, second, or third aspect, each of the first, second, or third substances is one of the following: a cell, a protein, a nucleic acid, and / or a small molecule.

[0022] As one possible implementation of the first, second, or third aspect, the bonding position toward which the pulsed acoustic beam is directed includes: the position of the carrier corresponding to the second or third substance to which the pulsed acoustic beam is directed.

[0023] As one possible implementation of the first, second, or third aspect, the pulsed acoustic beam is positioned at an acute angle to the carrier, such that the pulsed acoustic beam is directed toward the position of the carrier corresponding to the second or third substance to which it is bonded.

[0024] A fourth aspect of this application provides a method for separating a second substance bound to a first substance, comprising:

[0025] The affinity between the first substance and the second substance is analyzed using any of the methods described in the first, second, or third aspects, wherein the magnitude of the affinity is characterized by the number of pulses in the pulsed acoustic beam.

[0026] The carrier containing the first substance, which is to be separated from the second substance, is placed in a liquid environment;

[0027] The ultrasonic device is placed in a liquid environment and driven to generate a pulsed sound beam in the liquid environment. The number of pulses of the pulsed sound beam is adapted to the affinity between the first substance and the second substance and acts on the bonding position between the target second substance and the first substance to achieve the separation of the target second substance and the first substance.

[0028] The fifth aspect of this application provides a method for screening a target second substance, comprising:

[0029] Using the method described in any of the first, second, or third aspects, characterize the magnitude of the affinity of the second substance and the third substance for the first substance;

[0030] Based on the order of affinity, the second substance in the desired order is selected as the target substance.

[0031] A sixth aspect of this application provides a cell processing apparatus for implementing the method described in any of the first, second, or third aspects, the processing apparatus comprising:

[0032] A liquid environment for placing a carrier, wherein a first substance is attached to the carrier, and at least a portion of the first substance is bound to a second substance by affinity;

[0033] A supersonic device, wherein the supersonic device is driven to generate a pulsed acoustic beam in the liquid environment, and the pulsed acoustic beam acts on the bonding site between a first substance and a second substance to which it is bonded.

[0034] An image acquisition device is used to acquire continuous images including a first substance, for the purpose of counting the number of times the second substance detaches from the first substance under a certain number of pulses.

[0035] As a possible implementation of the sixth aspect, it further includes: a stage for carrying the liquid environment vessel, which is movable horizontally; or a first robotic arm equipped with a probe having the ultrasonic device, which allows the ultrasonic device to be moved spatially.

[0036] In summary, this application utilizes pulsed acoustic beams to quantitatively or qualitatively measure the affinity between cells or between cells and proteins. The affinity characterized by this method corresponds to minimal cell damage; that is, when the characterized affinity is applied to cells, the cells maintain good viability. Furthermore, this method provides highly accurate affinity analysis, enabling the measurement and analysis of affinity at the single-cell level. Based on the obtained affinity, this application can also be applied to the dissociation of target cells or proteins, or the selection of target cells or proteins (e.g., selecting a protein with the strongest affinity for a cell), for further subsequent research. Attached Figure Description

[0037] Figure 1 These are images and diagrams illustrating jet and secondary flow phenomena.

[0038] Figure 2a and Figure 2b This is a diagram of the acoustic beam generated in a liquid environment according to an embodiment of this application;

[0039] Figure 3 This is a timing diagram of the output signal of the driving device of the ultrasonic device provided in this application embodiment:

[0040] Figure 4 This is a schematic diagram of the driving device for the ultrasonic device provided in the embodiments of this application;

[0041] Figures 5a-5e This is a schematic diagram of the cell processing device provided in the second embodiment of this application;

[0042] Figure 6 This is a schematic diagram of the probe of the replaceable ultrasonic device with different characteristics in this application;

[0043] Figure 7 This is a schematic diagram of the dimensions of a special ultrasonic device;

[0044] Figure 8 This is a data graph plotted using the scheme provided in the embodiments of this application for affinity experiment four.

[0045] Figure 9 This is a data graph plotted using the scheme provided in the embodiments of this application for affinity experiment five.

[0046] The attached figures are labeled as follows: 1-stage, 2-robotic arm, 21-first rotating part, 22-probe, 23-ultrasonic device, 3-second robotic arm, 31-linear motor, 32-second rotating part, 33-pipette, 4-image acquisition device, 41-lens, 42-eyepiece, 5-liquid supply channel, 51-liquid outlet of the liquid supply channel.

[0047] It should be understood that the dimensions and shapes of the blocks in the above structural diagrams are for reference only and should not constitute an exclusive interpretation of the embodiments of the present invention. The relative positions and inclusion relationships between the blocks presented in the structural diagrams are only schematic representations of the structural relationships between the blocks, and are not intended to limit the physical connection methods of the embodiments of the present invention. Detailed Implementation

[0048] The technical solutions provided in this application will be further described below with reference to the accompanying drawings and embodiments. It should be understood that the system architecture and business scenarios provided in the embodiments of this application are mainly for illustrating possible implementations of the technical solutions of this application and should not be construed as the sole limitation on the technical solutions of this application. Those skilled in the art will recognize that the technical solutions provided in this application are equally applicable to similar technical problems as system architectures evolve and new business scenarios emerge.

[0049] It should be understood that the affinity analysis scheme provided in the embodiments of this application includes affinity analysis methods, applications, and cell processing devices. The applications include methods for separating a second substance bound to a first substance and methods for screening target second substances, such as applications for the dissociation of cells and proteins, and applications for screening target cells or proteins. Since these technical solutions solve problems based on the same or similar principles, some repetitions may not be repeated in the following descriptions of specific embodiments. However, it should be considered that these specific embodiments have mutual references and can be mutually referenced and combined.

[0050] 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 this application belongs. In case of any inconsistency, the meaning set forth in this specification or derived from the content described herein shall prevail. Furthermore, the terminology used herein is for the purpose of describing embodiments of this application only and is not intended to limit this application. To accurately describe the technical content of this application and to accurately understand the invention, the following explanations or definitions of the terms used in this specification are provided before describing specific embodiments:

[0051] 1) Affinity: Affinity usually refers to the mutual attraction or binding ability between different molecules or structures. For example, the affinity between cells, the affinity between cells and proteins, the affinity between proteins, the affinity between proteins and nucleic acids (such as DNA or RNA), the affinity between cells or proteins and small molecules, etc.

[0052] For the sake of simplicity in the following description, the affinity analysis protocol of this application will be described using the affinity between a cell and a protein bound to it as an example. It should be understood that the affinity analysis protocol of this application can be applied to various types of affinity, whether listed above or not.

[0053] 2) Cell confluence: This refers to the degree to which cells cover the surface of a culture medium (such as a petri dish or plate), usually expressed as a percentage. Cell confluence reflects the growth and distribution of cells on the culture medium. Generally, 80% confluence is a common standard for cells to be in an optimal growth state, at which point the number of cells is moderate, neither overcrowded nor too sparse, suitable for various experimental operations.

[0054] 3) Ultrasonic Device: A high-frequency resonator, which can be a device that generates mechanical vibration by applying voltage based on the piezoelectric effect. In this application, a piezoelectric resonator that generates ultrasonic waves of not less than 0.5 GHz (GHz is equivalent to gigahertz) during operation is used. Preferably, it is a piezoelectric resonator that generates ultrasonic waves of not less than 1 GHz and not more than 30 GHz during operation, for example, it can be 2 GHz to 2.5 GHz. Such piezoelectric resonators can be, for example, surface acoustic wave (SAW) devices, bulk acoustic wave (BAW) devices, etc. For example, when it is BAW, it can be a thin-film bulk acoustic wave resonator (FBAR), a solid-state assembled resonator (SMR), or a Lamb wave resonator (LWR). For ease of description, the piezoelectric resonator that can generate ultrasonic waves of not less than 0.5 GHz will be referred to as an ultrasonic device from now on.

[0055] 4) Jet phenomenon: This is a phenomenon that occurs when the sound waves from an ultrasonic device act on a liquid. The regional vibration generated at the working interface of the ultrasonic device can form a traveling wave in the liquid and exert a continuous thrust (i.e., volume force) on the local liquid in the liquid environment, causing at least a portion of the liquid to move in a straight line along the direction of sound wave propagation. This phenomenon of straight-line motion is called jet phenomenon.

[0056] Secondary flow phenomena, including eddies and thermal backflow, are another phenomenon generated when ultrasonic devices act on liquids. They include eddies (or micro vortices) caused by local circulation generated by the jet driving the liquid, and thermal backflow generated by the heating of ultrasonic devices.

[0057] The jet phenomenon and secondary flow phenomenon can be found in [reference needed]. Figure 1 The images and diagrams shown. Figure 1 The diagram shown illustrates the use of eddies to capture the first substance.

[0058] 5) Acoustic Beam: In this application, the acoustic beam is generated based on an ultrasonic device. In this application, the acoustic beam is a unique type of jet phenomenon, characterized by the fluid moving at high speed along the direction of sound wave propagation and exhibiting a thin cylindrical shape within its path. Before significant attenuation, the acoustic beam and the surrounding liquid are essentially in a laminar flow state, with low mixing. For example... Figure 2a and Figure 2b An image of the resulting acoustic beam, captured by a high-speed camera, is shown. Figure 2a and Figure 2b It can be seen that the acoustic beam has focusing properties and is no longer like... Figure 1 A cluster of jets, and Figure 2a and Figure 2b It is clearly not visible in the middle Figure 1 Secondary flow phenomena in [the context of something].

[0059] The acoustic beam generation conditions in this embodiment are: the ultrasonic device is driven to operate for at least one driving cycle at a first power, and each driving cycle includes a first stage and a second stage (see...). Figure 3 The drive unit outputs a signal portion that, in the first stage, drives the ultrasonic device to generate a 0.5-30 GHz ultrasonic effect in a liquid environment, and in the second stage, stops driving the ultrasonic device. The first power is sufficient to enable the ultrasonic device to generate a focused columnar acoustic beam perpendicular to the solid-liquid interface within the first stage. The duration of the first stage at the first power is less than a threshold, or the duty cycle of the first stage within one drive cycle is less than a threshold, to suppress the temperature of the ultrasonic device within a controllable temperature threshold and to prevent the ultrasonic device from generating secondary flows (including eddies and backflows) in the liquid environment.

[0060] The intensity of the acoustic beam is related to the duration of the first stage (or the duty cycle within a driving cycle) and the intensity of the first power. The duration and power of the first stage determine the amount of energy input to the ultrasonic device within a driving cycle. For example, a shorter first stage duration better suppresses temperature and secondary current, but also results in a lower acoustic beam intensity (a shorter first stage duration means a shorter time for energy accumulation). Conversely, a higher first power leads to a higher acoustic beam intensity and consequently a higher temperature rise in the ultrasonic device. Therefore, applying a higher first power and a shorter first stage not only yields a higher intensity acoustic beam but also effectively suppresses temperature and secondary current. Thus, when real-time control of the acoustic beam intensity is required, the preferred approach is to use a shorter, fixed first stage (e.g., the duty cycle of the first stage within the cycle is constant) and control the intensity of the generated acoustic beam by applying different amounts of first power. The alternative is to apply the same initial power and adjust different initial stage values ​​(such as the duty cycle of the initial stage value within the period) to control the intensity of the generated acoustic beam, or to adjust the initial stage value and the initial power simultaneously to control the intensity of the generated acoustic beam.

[0061] Once the liquid environment is determined and the ultrasonic device is selected, the relationship between the acoustic beam intensity and control parameters (first power and / or the periodic signal composed of the first and second stages) can be calibrated. In one scenario, if the periodic signal is a constant (i.e., the first and second stages are constant), calibrating the acoustic beam intensity and the first power can yield the magnitude of the first power and the acoustic beam intensity.

[0062] 6) Regarding the first stage within the drive cycle: In the first stage of the drive cycle, the drive 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 drive unit will output 1000 signals during this first stage. The power applied during the first stage will be applied to these 1000 signals to drive the ultrasonic device. In the second stage of the drive cycle (corresponding to...) Figure 3 (Switch signal low level) No drive signal output.

[0063] In certain cases, such as when the first phase is executed only once and then the process ends, this is equivalent to or considered as executing only one drive cycle in this application, and therefore this situation is also within the scope of protection of this application.

[0064] 7) Driving device for special ultrasonic devices: such as Figure 4 One embodiment is shown, including a control unit and a drive unit, the drive unit comprising a signal generator and a power amplifier. Its signal output principle can be found in [reference needed]. Figure 3 As shown.

[0065] The signal generator is used to generate high-frequency signals. The frequency of the original high-frequency signal it generates is the same as or approximately the operating frequency (or natural frequency) of the ultrasonic device used as a load. The waveform of the signal generator can be a rectangular wave (e.g.,...). Figure 3 The signals shown include square waves, sawtooth waves, sharp pulses (or triangular waves), stepped waves, sine waves, or half-waves. The output signal of the corresponding drive unit is modulated into sawtooth waves, sharp pulses (or triangular waves), stepped waves, sine waves, or half-waves.

[0066] A power amplifier is used to amplify the signal to be output so that it can drive a high-performance ultrasonic device.

[0067] The control unit can be a switching power supply, which controls the output switching signal. This switching signal can be a periodic signal, with a high level corresponding to the switching on phase (such as the conduction phase of the switching power supply's switching transistor). Figure 3 In the first stage, the switch-off stage corresponds to a low level, which is... Figure 3 The second stage involves controlling the operating voltage or amplification factor input to the power amplifier to achieve different output powers, which is the first power mentioned above. The first power can also be understood as the average energy density input to the ultrasonic device during one drive cycle (which consists of a first stage and a second stage). Since the energy level is related to power and time, the amount of energy input to the ultrasonic device during one drive cycle is related to the level of the first power, the duration of the first stage, or its duty cycle.

[0068] 8) Pulsed Acoustic Beam. When the driving device of an ultrasonic device outputs signals for each driving cycle, due to the viscous properties of the liquid itself, when the duration of the second stage (corresponding to the low-level signal) within the driving cycle is short, below a certain duration, the acoustic beam may still exhibit continuous characteristics. When the second stage exceeds a certain duration (e.g., the second stage accounts for 50%, 70%, etc.), the acoustic beam will exhibit obvious interval characteristics. In this embodiment, an acoustic beam with obvious interval characteristics is referred to as a pulsed acoustic beam. Pulsed acoustic beams are easier to quantitatively control or apply to metrology, etc.

[0069] In some embodiments, pulsed acoustic beams with different driving signal parameters can be generated by adjusting certain driving signal parameters (e.g., set power, set signal duty cycle). These pulsed acoustic beams can be used to measure and analyze affinity and the corresponding cell activity. Pulsed acoustic beams with driving signal parameters that maintain high cell activity can be selected to measure and analyze the affinity between cells and bound proteins.

[0070] The acoustic beam and related technologies for driving ultrasonic devices to generate acoustic beams, as described above, can also be found in the relevant description in Chinese patent application number CN20108322079.

[0071] The affinity analysis scheme provided in this application can control a special ultrasound device to generate pulsed acoustic beams in a liquid. Based on the pulsed acoustic beams acting on cells bound to proteins, it can be used to compare the number of different proteins detached from cells at a set number of pulses for qualitative affinity comparison. Alternatively, it can be used to pulse a specific protein on a single cell, recording the number of pulses of the acoustic beam used when the protein detaches from the cell as a quantitative marker of affinity. This scheme can be applied to the analysis of different affinities of different types of proteins, and can be further applied to the use of pulsed acoustic beams to detach proteins bound to cells, or to the selection of target proteins based on affinity for further research. For example, it can be applied to antibody affinity screening, safety testing of surface coatings for clinical medical materials, drug affinity screening, and validation of genetically modified cells.

[0072] The solutions provided in this application will now be described in detail with reference to the accompanying drawings and embodiments.

[0073] The first embodiment of this application provides a method for analyzing affinity, including:

[0074] S10: The carrier with the first substance attached is placed in a liquid environment, wherein a portion of the first substance is bound to the second substance by affinity, and a portion of the first substance is bound to the third substance by affinity.

[0075] S12: Place the ultrasonic device in a liquid environment and make the relative position of the ultrasonic device and the carrier adjustable;

[0076] S14: Drive the ultrasonic device to generate a pulsed sound beam in the liquid environment, and continuously adjust the relative position of the ultrasonic device and the carrier so that the pulsed sound beam acts on the bonding position of each first substance with the second or third substance it is bonded to.

[0077] S16: Acquire a continuous image including the first substance, and count the number of times the second and third substances detach from the first substance under a set number of pulses, in order to compare the affinity of the second and third substances for the first substance.

[0078] One specific example of the first embodiment involves coating the bottom of a culture dish with a type of cell, then culturing multiple proteins that bind to these cells. These proteins correspond to different affinities. A continuously moving ultrasonic device delivers a pulsed acoustic beam to a specific area at the bottom of the culture dish. After a predetermined number of pulses (or equivalent time), the ultrasonic device is stopped, and the number of different proteins separated from the cells at the bottom of the culture dish is recorded. Based on the ranking of these numbers, it is determined which protein the cells have a high affinity for and which has a low affinity for. A higher number of proteins separated into the suspension indicates a lower affinity for that protein and makes it easier to detach.

[0079] A more specific example is this: the bottom of the culture dish can be coated with a certain type of cancer cell, and the various proteins can be different target proteins. Using the above method, it's possible to analyze which target protein has a stronger affinity for the cancer cell. Furthermore, in biopharmaceutical manufacturing, target proteins with a greater affinity for cancer cells can be selected and subjected to gene testing or drug delivery, ultimately leading to the quantification of cancer drugs based on the selected target proteins.

[0080] The second embodiment of this application provides a method for analyzing affinity, including:

[0081] S20: The carrier with the first substance attached is placed in a liquid environment, wherein the second substance is bound to the first substance by affinity;

[0082] S22: Place the ultrasonic device in a liquid environment and make the pulsed sound beam of the ultrasonic device able to act on the junction of a first substance and a second substance.

[0083] S24: Drive the ultrasonic device to generate a pulsed acoustic beam at the junction of the first and second substances in the liquid environment;

[0084] S26: Acquire a continuous image including the first substance, determine the number of pulses of the pulsed acoustic beam when the second substance detaches from the first substance, the number of pulses being used to characterize the affinity between the first substance and the second substance.

[0085] One specific example of the second embodiment is as follows: A type of cell is coated on the bottom of a culture dish, and then a protein is cultured to bind to the cells. A pulsed beam of ultrasound is used to strike a single cell at the bottom of the culture dish. When the protein is separated from the single cell coated on the bottom of the culture dish, the number of pulses used is recorded to characterize the affinity.

[0086] This allows for the stripping of proteins from each cell based on the determined affinity (number of pulses), yielding a large quantity of the protein for subsequent research, such as protein structure studies in structural biology (requiring multiple studies of the same protein using different tests).

[0087] The third embodiment of this application provides a method for analyzing affinity, including:

[0088] S30: The carrier with the first substance attached is placed in a liquid environment, wherein the second substance is bound to the first substance by affinity;

[0089] S32: Place the ultrasonic device in a liquid environment and make the relative position of the ultrasonic device and the carrier adjustable;

[0090] S34: Drive the ultrasonic device to generate a pulsed sound beam in the liquid environment, and continuously adjust the relative position of the ultrasonic device and the carrier so that the pulsed sound beam acts on the bonding position of each first substance and the second substance it is bonded to.

[0091] S36: Acquire a continuous image including the first substance, and count the number of times the second substance detaches from the first substance under a set number of pulses. The value of the set number of pulses divided by the number of times the second substance detaches from the first substance is used to characterize the affinity between the second substance and the first substance.

[0092] In some embodiments, the first substance, the second substance, or the third substance may each be any of the following: cells, proteins, nucleic acids, and / or small molecules.

[0093] Therefore, the aforementioned affinity can be cell-to-cell affinity, cell-to-protein affinity, protein-to-protein affinity, protein-to-nucleic acid (e.g., DNA or RNA) affinity, cell or protein-to-small molecule affinity, and so on.

[0094] In some embodiments, the bonding position toward which the pulsed acoustic beam is directed includes the position of the carrier corresponding to the second or third substance to be bonded. In some embodiments, the carrier, such as a glass slide or petri dish, may have a first substance coated on its surface through cultivation or incubation. Specifically, the position corresponding to the second or third substance to be bonded, i.e., the target position, is the position facing the carrier.

[0095] In some embodiments, the pulsed acoustic beam is positioned at an acute angle to the carrier, directed towards the location on the carrier corresponding to the bonded second or third material. This angled orientation towards the target location provides a slanted peeling force, causing the second or third material bonded to the first material at the target location to be peeled off (while the first material at the target location remains on the carrier).

[0096] This application also provides applications following affinity analysis, some of which are listed below:

[0097] The fourth embodiment of this application provides a method for separating a second substance bound to a first substance, including:

[0098] S40: The affinity between the first substance and the second substance is analyzed using the above-mentioned affinity analysis method, wherein the magnitude of the affinity is characterized by the number of pulses in the pulsed acoustic beam;

[0099] S42: Place the carrier containing the first substance with the second substance to be separated in a liquid environment; the second substance to be separated is referred to as the target second substance.

[0100] S44: The ultrasonic device is placed in a liquid environment, and the ultrasonic device is driven to generate a pulsed sound beam in the liquid environment. The number of pulses of the pulsed sound beam is adapted to the affinity between the first substance and the second substance, and acts on the bonding position between the target second substance and the first substance to achieve the separation of the target second substance and the first substance.

[0101] The fifth embodiment of this application provides a method for screening out a target second substance. In this embodiment, each second substance is combined with multiple substances, such as a second substance and a third substance. The method includes:

[0102] S50: Using the above-described affinity analysis method, characterize the magnitude of the affinity between the second substance and the third substance and the first substance, respectively;

[0103] S52: Sort the substances according to their affinity and select the second substance in the desired sorting position as the target substance. For example, if the second substance has the highest affinity, the target substance with the highest affinity to the first substance should be selected, i.e., the second substance. Further research and processing can then be carried out based on the second substance.

[0104] Accordingly, this application also provides a cell processing device for implementing the above-mentioned affinity analysis method or its application, the processing device comprising:

[0105] A liquid environment for placing a carrier, wherein a first substance is attached to the carrier, and at least a portion of the first substance is bound to a second substance by affinity;

[0106] A supersonic device, wherein the supersonic device is driven to generate a pulsed acoustic beam in the liquid environment, and the pulsed acoustic beam acts on the bonding site between a first substance and a second substance to which it is bonded.

[0107] An image acquisition device is used to acquire continuous images including a first substance, for the purpose of counting the number of times the second substance detaches from the first substance under a certain number of pulses.

[0108] In some embodiments, it further includes: a stage for carrying the liquid environment vessel, which is movable horizontally; or a first robotic arm equipped with a probe having the ultrasonic device, which allows the ultrasonic device to be moved spatially.

[0109] For example, by adjusting the first robotic arm or the horizontal two-axis displacement stage where the stage is located, the position of the pulsed sound beam generated by the ultrasonic device can be adjusted so that the first substances on the carrier can move continuously, thereby achieving regional stripping of the second or third substances bonded to the first substances.

[0110] In some embodiments, the liquid environment may be a container such as a cell culture dish containing liquid. In some embodiments, the carrier of the first substance may be the inner wall (including the sidewall or bottom) of a container such as a cell culture dish. In other embodiments, the carrier may be a carrier such as a glass slide, which can be placed in a container such as a cell culture dish containing liquid.

[0111] In some embodiments, a specific implementation of the cell processing device may be as follows: Figures 5a-5e As shown, specifically, the processing device may include:

[0112] Stage 1, on which cell culture dishes and other containers can be placed, the inner surface of the container can be coated with a first substance as a carrier, or a glass slide or other carrier coated with the first substance can be placed inside the container, and a second substance or a third substance can be attached to the first substance.

[0113] The first robotic arm 2 has a probe 22 mounted on its movable end. The probe 22 is equipped with a special ultrasonic device 23. The posture of the probe 22 above the stage 1 can be adjusted by the first robotic arm 2, thereby adjusting the posture of the special ultrasonic device 23.

[0114] The ultrasonic device 23 can extend into the liquid environment and generate a pulsed acoustic beam that peels off a second or third substance bound to the first substance from a carrier coated with a first substance in the liquid environment.

[0115] In some embodiments, the first robotic arm 2 may be a three-axis robotic arm, which may be disposed beside the stage 1. In one embodiment, the movement of the three axes may be manually adjustable; in another embodiment, each axis of the three-axis robotic arm is equipped with a motor, which drives the movement of the movable end of the first robotic arm 2. In some embodiments, the movable end of the first robotic arm 2 is further equipped with a first rotating part 21, on which the probe 22 is mounted. In some embodiments, the first rotating part 21 may rotate along a horizontal axis. In some embodiments, the first rotating part 21 has a clamping part for clamping the probe 22. The first rotating part 21 may have its rotation angle manually adjustable, or its angle may be adjusted by a motor-driven rotation of the first rotating part 21. By combining the three-axis robotic arm with the first rotating part 21, the movement and rotation of the probe 22 are realized, thereby adjusting the ultrasonic device 23 at the end of the probe 22 to the desired position and desired orientation, so that the ultrasonic device 23 generates a pulsed sound beam at a desired angle in the liquid environment.

[0116] In some embodiments, a second robotic arm 3 may be included, the movable end of which is equipped with a pipette 33. The pipette 33 includes a pipette tube and a pipette tip located at the end of the pipette tube. The posture of the pipette 33 can be adjusted by the second robotic arm 3, thereby adjusting the posture of the pipette tip. The pipette tip can extend into the liquid environment to aspirate the detached suspension.

[0117] In some embodiments, the second robotic arm 3 may be a three-axis robotic arm. In one embodiment, the movement of the three axes can be manually adjusted; in another embodiment, each axis of the three-axis robotic arm is equipped with a motor, which drives the movement of the movable end of the second robotic arm 3. In some embodiments, the movable end of the second robotic arm 3 is also equipped with a linear motor 31 (or a T-axis motor), and the movable end of the linear motor 31 is equipped with a clamping part that clamps the body of the pipette 33. The linear motor 31 can extend or retract the pipette tip toward the target cell. In some embodiments, the movable end of the second robotic arm 3 is also equipped with a second rotating part 32, on which the linear motor 31 is mounted. Through the combination of the three-axis robotic arm, the second rotating part 32, and the linear motor 31, the movement of the pipette 33 is achieved, thereby adjusting the pipette tip at the end of the pipette 33 to the desired position.

[0118] In some embodiments, the stage 1 is horizontally movable. The stage 1 can be mounted on a horizontal guide rail, allowing it to be driven by a motor to move along the rail. For example, it can be mounted on a horizontal two-axis displacement stage, the movement of which can be manually adjusted, or each axis can be equipped with a corresponding motor for motor-driven movement. Thus, containers such as cell culture dishes on the stage 1 can be adjusted to the desired position.

[0119] In some embodiments, such as Figure 6 The probe 22 shown can be composed of two detachable parts: one part is for mounting to the rotating part 23 (e.g., to the clamping part of the rotating part 23), and the other part has the probe 22 end and is fitted with the special ultrasonic device 23. This allows for replacement of the probe 22 or the part with the probe 22 end as needed. Figure 6 The right-hand ultrasonic device 23 is smaller than the left-hand device. Under the same conditions, the diameter of the acoustic beam generated by the right-hand ultrasonic device 23 is much smaller than that generated by the left-hand ultrasonic device 23, which means it is more focused and can process a smaller unit area.

[0120] In some embodiments, the ultrasonic device 23 mounted on the end of the probe 22 can be located on both sides of the end of the probe 22, so that the probe 22 can be rotated 180 degrees along its axis and then clamped and fixed at the first rotating part 21, thereby realizing quick selection of the ultrasonic device 23 for use.

[0121] In some embodiments, such as Figure 5c As shown, at least a portion of the stage 1 is made of transparent material or has a hollowed-out design, such as... Figure 5d As shown, an image acquisition device 4 is disposed below the stage 1. The image acquisition device 4 includes at least one lens 41, which can be adjusted to face the stage 1. Thus, the image acquisition device 4 below can acquire images (since video is composed of frames, the images here include video) of the process and results of the pulsed acoustic beam, for analysis, or to control the working state of the ultrasonic device 23 (such as the number of pulses in the pulsed acoustic beam) and the position of the robotic arm's moving end based on the obtained image data or analyzed data.

[0122] In some embodiments, the image acquisition device 4 may be a microscope with multiple lenses 41, which are mounted on a rotating component to switch the working lens so that the working lens faces upward. The image acquisition device 4 may also include an eyepiece 42 or a display for observing the image acquired by the lens 41.

[0123] In some embodiments, the image acquisition device 4 may also be disposed beside or above the stage 1 (e.g., diagonally above), and the lens 41 of the image acquisition device 4 may be adjusted to face the stage 1 for image acquisition. In some embodiments, the image acquisition device 4 may also include multiple lenses 41 disposed in different positions (e.g., below, beside, or diagonally above the stage 1).

[0124] In some embodiments, such as Figure 5e As shown, it also includes a housing that at least accommodates the aforementioned stage 1, the first robotic arm 2 holding the probe 22, and the second robotic arm 3 holding the pipette 33, forming a relatively isolated space. This housing can be used as an incubator. The housing has an openable door for user operation (e.g., placing or removing containers from the stage 1), and the housing or door may have a transparent window for user observation of the interior.

[0125] In some embodiments, the space of the housing may also accommodate at least the lens 41 portion of the image acquisition device 4, so that the lens 41 is placed in a relatively clean housing.

[0126] In some embodiments, the interior of the enclosure further includes partitions to house necessary electrical equipment and equipment required for experiments. The electrical equipment may include at least one of the following: a power supply device, a drive device (such as a signal generator and power amplifier) ​​for driving the ultrasonic device 23, a pump and reservoir used in conjunction with the pipette 33, a drive device for driving the motors in the robotic arm or displacement stage, a control device (such as an industrial computer or computer), lighting equipment, etc. The control device is signal-connected to the drive device and other equipment, for example, to acquire images acquired by the image acquisition device 4, perform analysis and processing, and control the execution of corresponding components.

[0127] In some embodiments, the control device is further connected to an external display device and a human-machine interface (e.g., a keyboard or touchscreen). The control device receives user operation commands through the human-machine interface and controls the actions of other devices based on the commands, or reads a preset program based on the user's operation commands to automatically control other devices to cooperate in performing a series of operations (i.e., achieving automation). In some embodiments, the display device and human-machine interface may be located on the housing or exist independently of the housing. In some embodiments, a prominent position on the housing (e.g., ...) Figure 5e An emergency stop button can be installed on the upper right corner of the enclosure (as shown), which can be used to trigger a shutdown in an emergency.

[0128] In some embodiments, the robotic arm or displacement table may also be connected to a control device for direct operation, which may be mounted on the surface of the housing or on the outside of the housing.

[0129] In some embodiments, the ultrasonic device 23 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 a scalene pentagon. In other embodiments, it can be an oval, triangular, elliptical, rhomboid, semicircular, or any combination of shapes of any size polygon.

[0130] The area of ​​the ultra-ultrasonic device 23 (referring to the radial dimension, non-thickness dimension, i.e., the device interface dimension) is approximately 10-1,000,000 μm², preferably approximately 100-40,000 μm², and more preferably 1,000-10,000 μm². To provide a clearer understanding of the dimensions of the ultra-ultrasonic device 23, Figure 7 A dimensional diagram of the ultrasonic device 23 as an integrated acoustic resonator is shown. The dimensions of the ultrasonic device 23 are negatively correlated with the achievable resonant frequency; therefore, reducing the size can increase the resonant frequency. In other embodiments, the resonant frequency of the ultrasonic device 23 can also be changed by altering the shape of the device, such as the number of polygonal elements, the angles between adjacent sides, and the length of each side.

[0131] The diameter of the generated acoustic beam is related to the parameters of the ultrasonic device 23. The smaller the area (radial dimension or interface dimension) of the ultrasonic device 23, the smaller the diameter of the acoustic beam. Smaller microscales also result in better sound wave focusing. Specifically, under the same initial power application, using a smaller microscale for the ultrasonic device 23 results in a stronger acoustic beam, a smaller diameter of the acoustic beam, and a faster response speed (time to generate the acoustic beam).

[0132] The following describes the experiments performed using the above method for affinity analysis to illustrate the feasibility of this application.

[0133] The experimental materials used in this experiment are as follows:

[0134] 10kDa HA: This refers to HA molecules with a molecular weight of 10 kilodaltons. HA molecules (Hyaluronic Acid) are also known as hyaluronic acid. They are a type of polysaccharide molecule. In cell biology research, HA molecules are often used to mimic the extracellular matrix and promote cell adhesion and growth.

[0135] CD44 blocking antibody: CD44 is a biomarker on the membrane surface of tumor stem cells;

[0136] MDA-MB-231 cells: a type of human breast cancer cell;

[0137] PBS buffer: Buffer solutions are generally used as solvents. PBS buffer can be made from phosphate buffer.

[0138] The steps of this experiment are as follows:

[0139] S10: Coat cell culture dishes with 0.1 mg / mL HA molecules dissolved in PBS and incubate overnight at 4°C. Discard the waste liquid the next day for later use.

[0140] This step specifically includes: dissolving 0.1 mg / mL HA molecules in PBS, spreading the dissolved HA solution evenly on the surface of a cell culture dish; placing the coated culture dish at 4°C overnight (the low temperature of 4°C is to slow down the degradation of HA molecules and ensure that HA molecules can be uniformly and firmly fixed on the surface of the culture dish) to allow HA molecules to be fully fixed on the surface of the culture dish, completing the coating; the next day, discarding the waste liquid and preparing the culture dish for subsequent experiments.

[0141] The role of coating HA molecules is as follows: HA molecules can mimic the extracellular matrix, providing a surface conducive to cell adhesion and growth, making it easier for cells to adhere to the culture dish surface for growth and proliferation, such as the facilitating growth and proliferation of MDA-MB-231 cells described later on HA molecules. HA molecules can also bind specifically to specific cell surface receptors (such as the CD44 blocking antibody described later) in subsequent steps to study cell-HA molecule interactions.

[0142] S20: MDA-MB-231 cell extraction: When the confluence of MDA-MB-231 cells reaches 80% in another culture dish, calcein is added to the culture medium for staining for 20 minutes.

[0143] This step specifically includes: culturing and incubating MDA-MB-231 cells in culture medium in a culture dish, observing the cells using imaging equipment such as a microscope, assessing cell confluence, and culturing until the confluence reaches 80%. Then, adding calcein to the culture medium for staining for 20 minutes, thereby labeling the MDA-MB-231 cells for subsequent observation and analysis.

[0144] S30: After staining with MDA-MB-231, the cells were digested with trypsin and centrifuged at 1000 r / min for 5 min at room temperature.

[0145] This step specifically includes: adding an appropriate amount of trypsin solution to break down the junction proteins between cells, causing MDA-MB-231 cells to detach from the surface of their culture dish and form a single-cell suspension, thus restoring the previously adherent MDA-MB-231 cells to a suspended state. Then, the digested cell suspension can be transferred to a centrifuge tube, and the centrifuge parameters can be set to 1000 rpm for 5 minutes. After centrifugation, an MDA-MB-231 cell pellet will form at the bottom of the centrifuge tube.

[0146] S40: After centrifugation, discard the supernatant (mainly digestion solution and PBS buffer), add PBS buffer to resuspend the cells, and adjust the cell density by adding PBS buffer to dilute the MDA-MB-231 cell density to 1×10⁵ / well.

[0147] S50: At this point, the experimental group and the control group are set up as follows:

[0148] Experimental group: The MDA-MB-231 cell suspension at the above concentration was added to a cell culture dish coated with HA molecules, and MDA-MB-231 cells were incubated on the HA molecules. The incubated MDA-MB-231 cells bound to the HA molecules.

[0149] Control group: This group consisted of CD44 (0.5 mg / mL - 200 μL) blocking antibodies. 500 μL of a 10 μg / mL CD44 blocking antibody was added to an MDA-MB-231 cell suspension at the above concentration, and then the mixture was added to HA-coated cell culture dishes for MDA-MB-231 cell incubation. The CD44 blocking antibody blocks the specific adhesion of CD44 to HA; that is, the CD44 blocking antibody can specifically bind to HA, thereby preventing MDA-MB-231 cells from readily binding to HA.

[0150] S60: After incubation, a pulsed acoustic beam was generated using a special ultrasound device and applied to MDA-MB-231 cells that bound HA molecules. The affinity between MDA-MB-231 cells and HA was analyzed. The driving signal (power, signal period, and duty cycle were set to default values) corresponding to the pulsed acoustic beam was set to a constant value, and the number of pulses in the pulsed acoustic beam under this constant driving signal was used as the representation of affinity. The specific experimental details are as follows:

[0151] Experiment 1: Usability analysis of the experimental group: After incubation of the control group and the experimental group, a region of each culture dish was taken and a region peeling test (peeling against affinity) was performed using pulsed acoustic beam. Based on the experimental results (the number of remaining cells in the region of the culture dish: the experimental group was much larger than the control group), it was verified that the MDA-MB-231 cells in the experimental group were firmly bound to the HA molecules covering the cell culture dish, and the experimental group can be used for affinity analysis in this application.

[0152] Experiment 2, focusing on single-cell manipulation, tested affinity: The pulsed acoustic beam of the ultrasound device was tilted towards the surface of a single MDA-MB-231 cell bound to HA on the culture dish, and images were acquired using an electron microscope. The number of pulses of the pulsed acoustic beam used was recorded when the MDA-MB-231 cell detached from HA (HA still covered the surface of the culture dish). This number of pulses was used as a parameter to calibrate the affinity between MDA-MB-231 cells and HA.

[0153] Furthermore, by adjusting the relative positions of the movable ultrasound device and the culture dish, the above process was used again to calibrate the affinity parameters (number of pulses) between another MDA-MB-231 cell and HA. This process was repeated to measure the affinity parameters (number of pulses) between n MDA-MB-231 cells and HA, and the average value was taken as the final calibrated affinity parameter.

[0154] Experiment 3, focusing on a specific region, tested the affinity: The pulsed acoustic beam of the ultrasound device was tilted towards the surface of an MDA-MB-231 cell bound to HA on a culture dish. The ultrasound device or the culture dish was moved within the designated area so that the pulsed acoustic beam sequentially acted on each position within that area. The number of pulses used to complete the treatment of that area was recorded, and images before and after the test were obtained using an electron microscope. The reduction in the number of MDA-MB-231 cells bound to HA on the culture dish (i.e., the amount of detachment) was determined. The number of pulses corresponding to a single cell was calculated by dividing the number of pulses by the amount of cell reduction, which can be used as a parameter of the affinity between MDA-MB-231 cells and HA (i.e., the calculated number of pulses).

[0155] Experiment 4, not targeting a specific area, tests the affinity: The pulsed acoustic beam of the ultrasound device is tilted towards the surface of an MDA-MB-231 cell bound to HA on a culture dish. The ultrasound device or the culture dish is moved so that the pulsed acoustic beam acts sequentially on the surface of the cell bound to HA at different locations. The number of n pulses generated by the pulsed acoustic beam is recorded. Images before and after the test are obtained using an electron microscope to determine the reduction (i.e., detachment) of MDA-MB-231 cells bound to HA on the surface of the culture dish. The number of pulses corresponding to a single cell is calculated by dividing the number of pulses n by the reduction in the number of cells. This number can be used as a parameter of the affinity between MDA-MB-231 cells and HA (i.e., the calculated number of pulses).

[0156] Among them, such as Figure 8 The figure shows the relationship between the number of pulses in the pulsed acoustic beam and the remaining MDA-MB-231 cells on the surface of the culture dish, plotted using data obtained from Experiment 4. The two lines in the figure represent the fitting results using different algorithms.

[0157] In the above experiment, the method for determining the reduction (i.e., detachment) of HA-bound MDA-MB-231 cells on the culture dish based on images can be as follows: the cell image is identified and analyzed using fluorescence counting software based on the vibration image, the number of cells is counted, and the reduction (detachment into the suspension) of cells is calculated.

[0158] This can be a graph showing the relationship between the number of pulses and the remaining cell count under a set driving signal (power, signal period, and duty cycle are set values).

[0159] In addition, another experiment was conducted, which can be called Experiment Five: Three types of cells, such as granulocytes, monocytes, and lymphocytes, were treated according to steps S20-S30 above, but with different fluorescent staining. They were then mixed and added to cell culture dishes coated with HA molecules obtained in step S10 above for incubation (refer to step S50 above). Afterwards, Experiment Four in S60 was performed, and the number of remaining cells on the surface of the culture dish was collected after different pulse counts, thus allowing the calculation of the amount of cells detached after n pulses. Since the three types of cells have different detachment amounts (corresponding to detached areas) after n pulse counts, the order of the affinity of different cells for HA can be compared. After n pulse counts, the smallest detached area indicates the fewest detached cells and the greatest affinity of the cell for HA, and vice versa. Therefore, the desired (maximum or minimum) cells can be selected based on the binding force. Figure 9 The graph shown is plotted based on the data obtained in Experiment 5, depicting the relationship between the number of pulses in the pulsed acoustic beam and the area of ​​cells stripped from the surface of the culture dish.

[0160] In the several embodiments provided in this application, it should be understood that the disclosed apparatus and methods are not limited to the above embodiments and can be implemented in other ways. For example, the apparatus embodiments described above are merely illustrative. For instance, the division of units is only a logical functional division, and in actual implementation, there may be other division methods. For example, multiple units or components may be combined or integrated into another system, or some features may be ignored or not executed. Furthermore, the coupling or direct coupling or communication connection shown or discussed may be through some interfaces; the indirect coupling or communication connection between apparatuses or units may be electrical, mechanical, or other forms.

[0161] The units described as separate components may or may not be physically separate. The components shown as units may or may not be physical units; that is, they may be located in one place or distributed across multiple network units. Some or all of the units can be selected to achieve the purpose of this embodiment according to actual needs.

[0162] Furthermore, the terms "first, second, third, etc." or similar terms such as module A, module B, and module C used in the specification and claims are only used to distinguish similar objects and do not represent a specific ordering of objects. It is understood that, where permissible, a specific order or sequence may be interchanged so that the embodiments of this application described herein can be implemented in an order other than that illustrated or described herein.

[0163] In the above description, the labels of the steps involved, such as S10, S20, etc., do not mean that the steps will necessarily be executed. The order of the steps can be interchanged or executed simultaneously if permitted.

[0164] The term "comprising" as used in the specification and claims should not be construed as limiting itself to what follows; it does not exclude other elements or steps. Therefore, it should be interpreted as specifying the presence of the mentioned feature, integral, step, or component, but does not exclude the presence or addition of one or more other features, integrals, steps, or components, or groups thereof. Thus, the statement "device comprising means A and B" should not be limited to a device consisting solely of components A and B.

[0165] The terms "an embodiment" or "an embodiment" as used in this specification mean that a particular feature, structure, or characteristic described in conjunction with that embodiment is included in at least one embodiment of this application. Therefore, the terms "in one embodiment" or "in an embodiment" appearing throughout this specification do not necessarily refer to the same embodiment, but may refer to the same embodiment. Furthermore, in one or more embodiments, the particular features, structures, or characteristics can be combined in any suitable manner, as will be apparent to those skilled in the art from this disclosure.

[0166] Note that the above are merely preferred embodiments and the technical principles employed in this application. Those skilled in the art will understand that this application is not limited to the specific embodiments described herein, and various obvious changes, readjustments, and substitutions can be made without departing from the scope of protection of this application. Therefore, although this application has been described in detail through the above embodiments, this application is not limited to the above embodiments, and may include many other equivalent embodiments without departing from the concept of this application, all of which fall within the scope of protection of this application.

Claims

1. A method for analyzing affinity, characterized in that, include: A carrier containing a first substance is placed in a liquid environment, wherein a portion of the first substance is bound to a second substance by affinity, and a portion of the first substance is bound to a third substance by affinity. The ultrasonic device is placed in a liquid environment, and the relative position of the ultrasonic device and the carrier is adjustable; The ultrasonic device is driven to generate a pulsed sound beam in the liquid environment, and the relative position of the ultrasonic device and the carrier is continuously adjusted so that the pulsed sound beam acts on the bonding position of each first substance with the second or third substance it is bonded to. Acquire a continuous image including the first substance, and count the amount of the second and third substances detaching from the first substance under a set number of pulses, to compare the affinity of the second and third substances for the first substance.

2. A method for analyzing affinity, characterized in that, include: A carrier with a first substance attached is placed in a liquid environment, wherein a second substance is bound to the first substance by affinity. The ultrasonic device is placed in a liquid environment, and the pulsed sound beam of the ultrasonic device can act on the junction of a first substance and a second substance. The driving ultrasonic device generates a pulsed acoustic beam at the junction of the first and second substances in the liquid environment; Acquire a continuous image including the first substance, determine the number of pulses of the pulsed acoustic beam when the second substance detaches from the first substance, and the number of pulses is used to characterize the affinity between the first substance and the second substance.

3. A method for analyzing affinity, characterized in that, include: A carrier with a first substance attached is placed in a liquid environment, wherein a second substance is bound to the first substance by affinity. The ultrasonic device is placed in a liquid environment, and the relative position of the ultrasonic device and the carrier is adjustable; The ultrasonic device is driven to generate a pulsed acoustic beam in the liquid environment, and the relative position of the ultrasonic device and the carrier is continuously adjusted so that the pulsed acoustic beam acts on the bonding position of each first substance and the second substance it is bonded to. Acquire a continuous image including the first substance, and count the number of times the second substance detaches from the first substance under a set number of pulses. The value of the set number of pulses divided by the number of times the second substance detaches from the first substance is used to characterize the affinity between the second substance and the first substance.

4. The method according to any one of claims 1-3, characterized in that, The first substance, the second substance, or the third substance are each of the following: Cells, proteins, nucleic acids, and / or small molecules.

5. The method according to any one of claims 1-3, characterized in that, The bonding position of the pulsed acoustic beam includes the position of the carrier corresponding to the second or third substance to be bonded.

6. The method according to claim 5, characterized in that, The pulsed acoustic beam is positioned at an acute angle to the carrier, so that the pulsed acoustic beam is directed toward the position of the carrier corresponding to the second or third substance to which it is bonded.

7. A method for separating a second substance bound to a first substance, characterized in that: The affinity between the first substance and the second substance is analyzed using the method described in any one of claims 1-6, wherein the magnitude of the affinity is characterized by the number of pulses in the pulsed acoustic beam. The carrier containing the first substance, which is to be separated from the second substance, is placed in a liquid environment; The ultrasonic device is placed in a liquid environment and driven to generate a pulsed sound beam in the liquid environment. The number of pulses of the pulsed sound beam is adapted to the affinity between the first substance and the second substance and acts on the bonding position between the target second substance and the first substance to achieve the separation of the target second substance and the first substance.

8. A method for screening out a target second substance, characterized in that, include: Using the method described in any one of claims 1-6, characterize the magnitude of the affinity between the second substance and the third substance and the first substance, respectively; Based on the order of affinity, the second substance in the desired order is selected as the target substance.

9. A cell processing device for implementing the method according to any one of claims 1-6, characterized in that, The processing device includes: A liquid environment for placing a carrier, wherein a first substance is attached to the carrier, and at least a portion of the first substance is bound to a second substance by affinity; A supersonic device, wherein the supersonic device is driven to generate a pulsed acoustic beam in the liquid environment, and the pulsed acoustic beam acts on the bonding site between a first substance and a second substance to which it is bonded. An image acquisition device is used to acquire continuous images including a first substance, for the purpose of counting the number of times the second substance detaches from the first substance under a certain number of pulses.

10. The apparatus according to claim 9, characterized in that, Also includes: The stage that supports the liquid environment vessel is movable horizontally. or A first robotic arm equipped with a probe having the aforementioned ultrasonic device, which allows the ultrasonic device to move spatially.

Images