WO2024065683A1 - 一种细胞力学特性测量及分选系统、方法、终端及存储介质 - Google Patents

一种细胞力学特性测量及分选系统、方法、终端及存储介质 Download PDF

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WO2024065683A1
WO2024065683A1 PCT/CN2022/123288 CN2022123288W WO2024065683A1 WO 2024065683 A1 WO2024065683 A1 WO 2024065683A1 CN 2022123288 W CN2022123288 W CN 2022123288W WO 2024065683 A1 WO2024065683 A1 WO 2024065683A1
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cell
cells
interdigital transducer
sorting
acoustic field
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English (en)
French (fr)
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孟龙
周伟
郑海荣
牛丽丽
彭本贤
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Shenzhen Institute of Advanced Technology of CAS
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Shenzhen Institute of Advanced Technology of CAS
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Priority to PCT/CN2022/123288 priority Critical patent/WO2024065683A1/zh
Publication of WO2024065683A1 publication Critical patent/WO2024065683A1/zh
Priority to US19/004,464 priority patent/US20250137909A1/en
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1456—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
    • G01N15/1459—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals the analysis being performed on a sample stream
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/02—Investigating particle size or size distribution
    • G01N15/0205—Investigating particle size or size distribution by optical means
    • G01N15/0227—Investigating particle size or size distribution by optical means using imaging; using holography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1404—Handling flow, e.g. hydrodynamic focusing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1425—Optical investigation techniques, e.g. flow cytometry using an analyser being characterised by its control arrangement
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1429—Signal processing
    • G01N15/1433—Signal processing using image recognition
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1456—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/149—Optical investigation techniques, e.g. flow cytometry specially adapted for sorting particles, e.g. by their size or optical properties
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1006—Investigating individual particles for cytology
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1404—Handling flow, e.g. hydrodynamic focusing
    • G01N2015/142—Acoustic or ultrasonic focussing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N2015/1493—Particle size
    • G01N2015/1495—Deformation of particles

Definitions

  • the present application relates to neural regulation technology, and in particular to a cell mechanical property measurement and sorting system, method, terminal and storage medium.
  • cell mechanical properties have such an important influence on cell biological functions, it is crucial to accurately measure the mechanical properties of cells, whether to use cell mechanical properties to better understand their biological functions or to use external stimuli to change mechanical properties and thus change biological functions.
  • Cell mechanics has been proven to be a biophysical fingerprint that can distinguish cell phenotypes, reveal the process of aging or disease, and even detect and diagnose cell lesions.
  • the elastic modulus of healthy breast cells is more than twice that of breast cancer cells
  • the elastic modulus of benign breast tumor cells is 1.4-1.8 times that of malignant breast cancer cells. The decrease in cell elastic modulus can be used for cancer diagnosis.
  • red blood cells indicate cell aging and lesions.
  • the viscoelasticity and deformation ability of red blood cells can be used to distinguish between diseased and aged cells and healthy and young red blood cells, which puts high demands on the accurate measurement of cell elastic modulus. It can be seen that the cell Young's modulus is an inherent mechanical parameter of the cell.
  • One of the purposes of the present application is to provide a cell mechanical property measurement and sorting system, comprising: a first interdigital transducer, a second interdigital transducer, a cell elastic modulus calculation unit, a cell unit unit and a cell sorting unit, wherein:
  • the cells to be sorted are arranged in a straight line after passing through the standing wave acoustic field generated by the first interdigital transducer; the cells arranged in a straight line flow through the focused acoustic field generated by the second interdigital transducer, and the focused acoustic field generates a radiation force on the cells to cause the cells to deform; the cell elastic modulus calculation unit calculates the elastic modulus of the cells based on the information of the radiation force and the deformation; the cell positioning unit is used to determine the position of the cells, and the cell positioning unit is also used to selectively drive the interdigital transducer in the cell sorting unit according to the moving speed of the cells and the distance from the sorting unit with a designed delay time; the cell sorting unit triggers the corresponding interdigital transducer according to the elastic modulus to generate a planar acoustic field to sort the cells.
  • the cell elastic modulus calculation unit includes: a recording module and a computer, the recording module includes a high-speed camera or a microscope, the deformation of the cell is recorded by the recording module, and the computer calculates the elastic modulus of the cell based on the radiation force and the deformation information.
  • the cell localization unit comprises a photosensitive electrode.
  • the cell sorting unit comprises an array of interdigital transducers.
  • the second object of the present application is to provide a method for measuring and sorting cell mechanical properties, comprising the following steps:
  • the cells to be sorted are arranged in a straight line after passing through the standing wave acoustic field generated by the first interdigital transducer;
  • the cells arranged in a straight line flow through a focused acoustic field generated by a second interdigital transducer, and the focused acoustic field generates a radiation force on the cells to cause the cells to deform;
  • a delay time is designed to selectively drive the interdigital transducer
  • the elastic modulus triggers the corresponding interdigital transducer to generate a planar acoustic field to sort the cells.
  • the IDT includes the following preparation method: preparing the IDT and forming a PDMS cavity on the IDT.
  • the steps of preparing the interdigital transducer specifically include the following steps:
  • the photoresist is peeled off from the piezoelectric substrate on which the electrode layer is grown, thereby completing the preparation of the interdigital transducer.
  • the step of forming a PDMS cavity on the interdigital transducer specifically includes the following steps:
  • the upper surface of the IDT after exposure and development is bonded to the PDMS cavity for bonding.
  • the PDMS cavity is prepared by the following method:
  • the PDMS is injected into the mold, and the air bubbles in the PDMS are removed by vacuuming, and then the PDMS is cured by heating; the PDMS is peeled off from the cavity of the mold to obtain the PDMS cavity.
  • the focused acoustic field in the step where the cells arranged in a straight line flow through the focused acoustic field generated by the second interdigital transducer, the focused acoustic field generates a radiation force on the cells to cause the cells to deform, and the deformation of the cells is recorded by a high-speed camera or a microscope.
  • the step of calculating the elastic modulus of the cell according to the radiation force and the deformation information specifically includes the following steps:
  • the cell elastic modulus is characterized based on the simulation results.
  • the third object of the present application is to provide a terminal, the terminal comprising a processor and a memory coupled to the processor, wherein:
  • the memory stores program instructions for implementing the cell mechanical property measurement and sorting method
  • the processor is used to execute the program instructions stored in the memory to control cell sorting.
  • the fourth object of the present application is to provide a storage medium storing program instructions executable by a processor, wherein the program instructions are used to execute the cell mechanical property measurement and sorting method.
  • the cell mechanical properties measurement and sorting system, method, terminal and storage medium provided by the present application are arranged in a straight line after passing through the standing wave sound field generated by the first interdigital transducer; the cells arranged in a straight line flow through the focused sound field generated by the second interdigital transducer, and the focused sound field generates a radiation force on the cells to cause the cells to deform; the cell elastic modulus calculation unit calculates the elastic modulus of the cells based on the information of the radiation force and the deformation; the cell sorting unit triggers the corresponding interdigital transducer to generate a planar sound field to sort the cells according to the elastic modulus.
  • the cell mechanical properties measurement and sorting system, method, terminal and storage medium provided by the present application realize cell elasticity measurement and sorting through ultrasonic non-contact, thereby reducing damage to the cells.
  • FIG1 is a schematic diagram of the structure of the cell mechanical properties measurement and sorting system provided in Example 1 of the present application.
  • FIG2 is a flowchart of the steps of the method for measuring and sorting cell mechanical properties provided in Example 2 of the present application.
  • FIG3 is a flow chart of the steps for preparing an IDT provided in Example 2 of the present application.
  • FIG. 4 is a schematic diagram of a process for preparing an IDT provided in Example 2 of the present application.
  • FIG. 5 is a flow chart of the steps of forming a PDMS cavity on the IDT provided in Example 2 of the present application.
  • FIG6 is a schematic diagram of a cell being squeezed from a spherical shape to an ellipsoidal shape under the action of the ultrasonic radiation force generated by a standing wave acoustic field provided in Example 2 of the present application.
  • FIG. 7 is a schematic diagram of the terminal structure provided in Example 3 of the present application.
  • FIG8 is a schematic diagram of the structure of the storage medium provided in Example 4 of the present application.
  • first and second are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, a feature defined as “first” or “second” may explicitly or implicitly include one or more of the features. In the description of this application, the meaning of “plurality” is two or more, unless otherwise clearly and specifically defined.
  • Figure 1 is a structural schematic diagram of a cell mechanical property measurement and sorting system provided in Example 1 of the present application, including a first interdigital transducer 110, a second interdigital transducer 120, a cell elastic modulus calculation unit 130, a cell positioning unit 140 and a cell sorting unit 150.
  • the interdigital transducer given in the embodiment of the present application is formed by plating interdigital electrodes on a piezoelectric substrate.
  • a sine wave signal of a corresponding frequency is input, a surface wave signal of a corresponding frequency will be generated. Due to the characteristics of high frequency, miniaturization, and low energy loss, it can be applied to the manipulation of microscopic particles or cells.
  • the surface wave acoustic field can be regulated by regulating the morphology of the interdigital transducer to achieve the generation of a focused or planar acoustic field.
  • the cells can be deformed by the focused acoustic field to measure the mechanical properties of the cells.
  • the planar acoustic field can be selectively triggered by the mechanical property measurement structure to sort the cells.
  • the cell mechanical properties measurement and sorting system provided in this application works as follows:
  • Step S110 the cells to be sorted are arranged in a straight line after passing through the standing wave acoustic field generated by the first IDT 110. It can be understood that the surface wave acoustic field is regulated by regulating the IDT morphology, and the standing wave acoustic field generated makes the cells to be sorted arranged in a straight line.
  • Step S120 The cells arranged in a straight line flow through the focused acoustic field generated by the second IDT 120, and the focused acoustic field generates radiation force on the cells to deform the cells. It can be understood that by regulating the surface wave acoustic field by regulating the IDT morphology, the high-intensity focused acoustic field generated by the IDT 120 will generate radiation force on the cells, thereby achieving squeezing and deformation of the cells.
  • Step S130 the cell elastic modulus calculation unit 130 calculates the elastic modulus of the cell according to the radiation force and the deformation information.
  • the cell elastic modulus calculation unit 130 includes: a recording module 131 and a computer 132, the recording module 131 includes a high-speed camera or a microscope, the deformation of the cell is recorded by the recording module, and the computer 132 calculates the elastic modulus of the cell based on the radiation force and the deformation information.
  • Step S140 The cell positioning unit 140 can determine the position of the cell, and selectively drive the interdigital transducer in the cell sorting unit with a designed delay time according to the moving speed of the cell and the distance between the cell and the sorting unit.
  • the cell positioning unit 140 includes a photosensitive electrode 141 .
  • Step S150 the cell sorting unit 150 triggers the corresponding interdigital transducer to generate a planar acoustic field to sort the cells according to the elastic modulus.
  • the cell sorting unit 150 includes an array of interdigital transducers 151 .
  • the cell elastic modulus calculation unit 130 calculates the elastic modulus of the cells based on the radiation force and the deformation information, and the cell sorting unit 150 triggers the planar ultrasonic transducer to generate sound waves based on the elastic modulus.
  • the radiation force generated by the sound waves will push the soft cells to the upper end of the cavity; on the contrary, when hard cells flow through, the cell sorting unit 150 will not trigger the corresponding interdigital transducer based on the elastic modulus, and the hard cells will still flow in the original direction, thereby realizing cell sorting based on cell elasticity.
  • the cell mechanical property measurement and sorting system provided in the present application realizes cell elasticity measurement and sorting through ultrasonic non-contact, thereby reducing damage to cells.
  • Figure 2 is a step flow chart of the cell mechanical property measurement and sorting method provided in Example 2 of the present application, including the following steps S210 to S250. The implementation method of each step is described in detail below.
  • Step S210 the cells to be sorted are arranged into a straight line after passing through the standing wave acoustic field generated by the first interdigital transducer.
  • the preparation of the interdigital transducer provided in the present application is mainly made by plating interdigital electrodes and recording electrodes on a piezoelectric substrate.
  • 128° YX or X-cut double-sided polished lithium niobate is selected as the piezoelectric substrate.
  • the process of making the interdigital transducer mainly includes processes such as glue coating, photolithography, coating, and stripping.
  • the specific preparation method includes: preparing the interdigital transducer and forming a PDMS (polydimethylsiloxane) cavity on the interdigital transducer.
  • the steps of preparing the IDT specifically include the following steps S310 to S340 .
  • the implementation method of each step is described in detail below.
  • Step S310 coating a positive photoresist on the surface of the piezoelectric substrate.
  • the positive photoresist AZ5214 was spin-coated at 3000 rpm for 30 seconds on the surface of the completely cleaned piezoelectric substrate, and the piezoelectric substrate was placed on a 65°C hot plate and baked for 3 minutes.
  • the thickness of the photoresist was tested using a step profiler, and the thickness of the photoresist was approximately 1.5 ⁇ m, as shown in FIG4 (a).
  • Step S320 exposing and developing the piezoelectric substrate coated with the positive photoresist.
  • the prepared film is then covered on the surface of Figure 4 (a) for exposure.
  • the patterned portion is opaque, the non-patterned portion is translucent, and the portion through which light is transmitted will solidify.
  • the solidified portion is dissolved, and the non-solidified portion will not be dissolved.
  • the development forms a pattern as shown in Figure 4 (b).
  • Step S330 performing magnetron sputtering on the piezoelectric substrate after the exposure and development processes to grow a metal electrode layer or a photosensitive electrode layer on the surface of the piezoelectric substrate.
  • magnetron sputtering is performed on the substrate after the pattern transfer is completed, so as to grow a metal electrode or photosensitive electrode layer with a thickness of about 200 nm, as shown in FIG. 4 (c).
  • Step S340 stripping the photoresist from the piezoelectric substrate on which the electrode layer is grown, thereby completing the preparation of the interdigital transducer.
  • the substrate on which the electrode is grown is placed in an acetone solution, and the photoresist is stripped off by ultrasonic vibration of an ultrasonic cleaning machine to complete the fabrication of the interdigital transducer, as shown in FIG4(d).
  • the step of forming a PDMS cavity on the IDT specifically includes the following steps S410 to S440 .
  • the implementation method of each step is described in detail below.
  • Step S410 pre-processing the IDT.
  • Step S420 coating a negative photoresist on the pre-treated surface of the IDT and drying the photoresist.
  • a coating machine is used to spin-coat SU-8 (50) negative photoresist on the surface of the pretreated IDT, wherein the coating parameters are as follows: 3000 rpm, 30 s, and the thickness of SU-8 (50) is about 50 ⁇ m; after coating, the IDT is horizontally placed on a 95° C. heating plate for 45 min to allow the solvent in the photoresist to evaporate, so as to enhance the adhesion between the photoresist and the IDT, thereby obtaining the pattern shown in FIG4 (e).
  • Step S430 performing exposure and development processing on the dried IDT.
  • a film with a pattern made is placed on an IDT with photoresist spin-coated, and the photoresist is exposed by an exposure machine with an exposure dose of 200 mJ/ cm2 . After exposure, the film is heated at 65°C for 1 min; and at 95°C for 3-5 min. The exposed IDT is soaked in a developer, the photoresist in the unexposed area is dissolved, and the photoresist in the exposed area is retained, and the pattern in Figure 4 (h) is obtained.
  • Step S440 bonding the upper surface of the IDT after the exposure and development process to the PDMS cavity for bonding.
  • the PDMS cavity is prepared by the following method: injecting PDMS into a mold, vacuuming to remove bubbles in the PDMS, and then heating to solidify the PDMS; peeling the PDMS from the cavity of the mold to obtain the PDMS cavity.
  • PDMS glue A and glue B are mixed in a mass ratio of 10:1, poured into a mold, and vacuum is applied to remove bubbles in the PDMS. Finally, the culture dish is placed in an oven at 80°C for 30 minutes to solidify the PDMS, as shown in Figure 4 (g); the PDMS cavity is peeled off from the mold to obtain the result shown in Figure 4 (h).
  • the fabricated IDT and PDMS cavity were subjected to plasma treatment at a power of 150 W for 70 seconds, and then the PDMS cavity was bonded to the IDT and baked in an oven at 80° C. for 20 minutes, thereby obtaining the fabricated IDT for experiment as shown in FIG4 (I).
  • Step S220 the cells arranged in a straight line flow through a focused acoustic field generated by a second interdigital transducer, and the focused acoustic field generates a radiation force on the cells to cause the cells to deform.
  • the focused acoustic field in the step where the cells arranged in a straight line flow through the focused acoustic field generated by the second interdigital transducer, the focused acoustic field generates a radiation force on the cells to cause the cells to deform, and the deformation of the cells is recorded by a high-speed camera or a microscope.
  • Step S230 Calculate the elastic modulus of the cell according to the radiation force and the deformation information.
  • the step of calculating the elastic modulus of the cell based on the radiation force and the deformation information specifically includes the following steps S231 to S233, and the specific implementation method of each step is described in detail below.
  • Step S231 Obtain cell deformation information.
  • a low-intensity standing wave acoustic field is first used to arrange the cells at the acoustic wave node position, and then a high-intensity focused standing wave ultrasound is used to act on the cells.
  • the ultrasonic radiation forces on both ends of the cell membrane are in opposite directions, thereby squeezing the cells and causing deformation.
  • the cell deformation mainly depends on the external force and the cell elastic modulus.
  • the external force is the main factor of cell deformation and is proportional to the cell deformation.
  • the cell under the action of the ultrasonic radiation force generated by the standing wave acoustic field, the cell is squeezed from a sphere to an ellipsoid.
  • the contour of the cell can be extracted and analyzed based on the image processing algorithm.
  • Step S232 Simulate the ultrasonic radiation force exerted on the cells.
  • the ultrasonic radiation force exerted on the cells is simulated based on the existing measurement results of the sound wave energy and information such as the cell sound velocity and density.
  • the incident standing wave acoustic field can be expressed as:
  • p a is the sound pressure amplitude
  • k o is the wave beam in the external medium
  • ⁇ 2 ⁇ f is the angular frequency
  • f is the ultrasonic frequency.
  • the sound wave propagation equation for the sound pressure p 1 is the Helmholtz equation:
  • ⁇ is the Laplace operator
  • c is the sound velocity of the medium
  • ultrasonic particle velocity can be expressed by the Euler formula:
  • ⁇ is the density of the medium.
  • i represents the average tensor in two-dimensional space
  • ⁇ > represents the time average within the sound wave cycle
  • Step S233 Characterize the cell elastic modulus based on the simulation results.
  • the neo-Hookean law is usually used.
  • the actual three-dimensional cell subjected to in-plane tensile force can be described by a two-dimensional model of the cell.
  • the elastic modulus formula derived from the neo-Hookean law is:
  • Es represents the elastic modulus of the cell membrane
  • ds0 and ds0 represent the infinitesimal inline unit of the cell membrane before and after deformation, respectively.
  • the elastic modulus of the cell can be calculated based on the radiation force and the deformation information.
  • Step S240 Designing a delay time to selectively drive the IDT according to the moving speed of the cell and the distance between the cell and the sorting unit.
  • the present application designs a cell positioning unit to determine the position of the cell, and designs a delay time to selectively drive the interdigital transducer in the cell sorting unit according to the movement speed of the cell and the distance between the cell and the sorting unit.
  • Step S250 The elastic modulus triggers the corresponding interdigital transducer to generate a planar acoustic field to sort the cells.
  • the cell elastic modulus calculation unit 130 calculates the elastic modulus of the cells based on the radiation force and the deformation information, and the cell sorting unit 150 triggers the planar ultrasonic transducer to generate sound waves based on the elastic modulus.
  • the radiation force generated by the sound waves will push the soft cells to the upper end of the cavity; on the contrary, when hard cells flow through, the cell sorting unit 150 will not trigger the corresponding interdigital transducer based on the elastic modulus, and the hard cells will still flow in the original direction, thereby realizing cell sorting based on cell elasticity.
  • the cell mechanical property measurement and sorting method provided in the present application realizes cell elasticity measurement and sorting through ultrasonic non-contact, thereby reducing damage to cells.
  • FIG7 is a schematic diagram of the terminal structure of Embodiment 3 of the present application.
  • the terminal 50 includes a processor 51 and a memory 52 coupled to the processor 51 .
  • the memory 52 stores program instructions for implementing the above-mentioned CT pancreatic tumor automatic segmentation method.
  • the processor 51 is used to execute program instructions stored in the memory 52 to control the automatic segmentation of CT pancreatic tumors.
  • the processor 51 may also be referred to as a CPU (Central Processing Unit).
  • the processor 51 may be an integrated circuit chip having signal processing capabilities.
  • the processor 51 may also be a general-purpose processor, a digital signal processor (DSP), an application-specific integrated circuit (ASIC), a field-programmable gate array (FPGA) or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components.
  • DSP digital signal processor
  • ASIC application-specific integrated circuit
  • FPGA field-programmable gate array
  • the general-purpose processor may be a microprocessor or the processor may also be any conventional processor, etc.
  • FIG 8 is a schematic diagram of the structure of the storage medium of Example 4 of the present application.
  • the storage medium of the embodiment of the present application stores a program file 61 that can implement all the above methods, wherein the program file 61 can be stored in the above storage medium in the form of a software product, including a number of instructions to enable a computer device (which can be a personal computer, server, or network device, etc.) or a processor (processor) to execute all or part of the steps of each implementation method of the present application.
  • a computer device which can be a personal computer, server, or network device, etc.
  • processor processor
  • the aforementioned storage medium includes: U disk, mobile hard disk, read-only memory (ROM, Read-Only Memory), random access memory (RAM, Random Access Memory), disk or optical disk and other media that can store program codes, or terminal devices such as computers, servers, mobile phones, tablets, etc.

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Abstract

本申请提供的细胞力学特性测量及分选系统、方法、终端及存储介质，待分选细胞经第一叉指换能器产生的驻波声场后排列成直线；呈直线排列的细胞流经第二叉指换能器产生的聚焦声场，聚焦声场对细胞产生辐射力以使细胞产生形变；细胞弹性模量计算单元根据辐射力和形变的信息计算得到细胞的弹性模量；细胞定位单元确定细胞的位置，并根据细胞的移动速度及与分选单元之间的距离，以设计延迟时间选择驱动细胞分选单元中的叉指换能器，细胞分选单元根据所述弹性模量触发对应的叉指换能器产生平面声场对所述细胞进行分选，上述分选系统、方法、终端及存储介质，通过超声非接触实现对细胞弹性测量以及分选，减少对细胞的损伤。

Description

一种细胞力学特性测量及分选系统、方法、终端及存储介质 技术领域
本申请涉及神经调控技术，特别涉及一种细胞力学特性测量及分选系统、方法、终端及存储介质。
背景技术
细胞力学特性对细胞生物学功能影响的研究，充分展示出力学因素的重要影响。由于细胞力学特性对细胞生物学功能有如此重要的影响，无论是利用细胞力学特性更好的理解其生物学功能，还是利用外界刺激改变力学特性从而改变生物学功能，准确地测量出细胞的力学特性都是至关重要的。细胞力学已被证明是一种生物物理指纹，能够区分细胞表型，揭示衰老或疾病的过程，甚至检测和诊断细胞病变。例如健康乳腺细胞的弹性模量是乳腺癌细胞弹性模量的2倍以上，良性乳腺肿瘤细胞的弹性模量是恶性乳腺癌细胞弹性模量的1.4-1.8倍。细胞弹性模量的下降可以用于癌症的诊断。红细胞较低的粘弹性和较差的变形能力预示着细胞的衰老和病变，可以根据红细胞的粘弹性和变形能力区分病变衰老的细胞和健康年轻的红细胞，这就对准确测量细胞弹性模量提出了很高的要求。可见，细胞杨氏模量作为细胞固有的力学参数。
现有的原子力显微镜、微管吸吮、磁镊技术等方法通量低、需要标记，限制了其在细胞力学测量方面的应用。声镊技术是利用声场中的物体对声波产生的反射、折射、吸收等效应引起的动量在声波与物体之间交换，从而产生力的作用，即超声辐射力(acoustic radiation force)。由于声镊具有无标记、非接触、高通量和普遍适用性等特点，在卵细胞活性评估方面具有重大的应用前景。
现有的原子力显微镜、微管吸吮、磁镊技术等方法通量低、需要标记，限 制了其在细胞力学测量方面的应用。
发明内容
鉴于此，有必要针对现有技术中存在的缺陷提供采用超声非接触的对细胞弹性测量且可实现对不同弹性模量细胞进行分选的分选系统及分选方法。
为解决上述问题，本申请采用下述技术方案：
本申请目的之一，提供了一种细胞力学特性测量及分选系统，包括：第一叉指换能器、第二叉指换能器、细胞弹性模量计算单元、细胞单位单元及细胞分选单元，其中：
待分选细胞经所述第一叉指换能器产生的驻波声场后排列成直线；呈直线排列的所述细胞流经所述第二叉指换能器产生的聚焦声场，所述聚焦声场对所述细胞产生辐射力以使所述细胞产生形变；所述细胞弹性模量计算单元根据所述辐射力和所述形变的信息计算得到细胞的弹性模量；所述细胞定位单元用于确定所述细胞的位置，所述细胞定位单元还用于根据所述细胞的移动速度及与所述分选单元之间的距离，以设计延迟时间选择驱动所述细胞分选单元中的叉指换能器；所述细胞分选单元根据所述弹性模量触发对应的叉指换能器产生平面声场对所述细胞进行分选。
在其中一些实施例中，所述细胞弹性模量计算单元包括：记录模块及计算机，所述记录模块包括高速相机或者显微镜，所述细胞产生形变由所述记录模块记录，所述计算机根据所述辐射力和所述形变的信息计算得到细胞的弹性模量。
在其中一些实施例中，所述细胞定位单元包括光敏电极。
在其中一些实施例中，所述细胞分选单元包括阵列的叉指换能器。
本申请目的之二，提供了一种细胞力学特性测量及分选方法，包括下述步骤：
待分选细胞经第一叉指换能器产生的驻波声场后排列成直线；
呈直线排列的所述细胞流经第二叉指换能器产生的聚焦声场，所述聚焦声场对所述细胞产生辐射力以使所述细胞产生形变；
根据所述辐射力和所述形变的信息计算得到细胞的弹性模量；
根据所述细胞的移动速度及与所述分选单元之间的距离，设计延迟时间选择驱动叉指换能器；
所述弹性模量触发对应的叉指换能器产生平面声场对所述细胞进行分选。
在其中一些实施例中，所述叉指换能器包括下述制备方法：制备叉指换能器及在所述叉指换能器上形成PDMS腔道。
在其中一些实施例中，在制备叉指换能器的步骤中，具体包括下述步骤：
在压电基底的表面涂覆正光刻胶；
将涂覆有正光刻胶的压电基底进行曝光和显影处理；
对曝光和显影处理后的压电基底进行磁控溅射，使其表面生长金属电极层或光敏电极层；
将生长电极层的压电基底剥离光刻胶，完成叉指换能器的制备。
在其中一些实施例中，在所述叉指换能器上形成PDMS腔道的步骤中，具体包括下述步骤：
对所述叉指换能器进行预处理；
在预处理后的所述叉指换能器的表面涂覆负光刻胶并烘干；
对烘干后的所述叉指换能器进行曝光和显影处理；
在曝光和显影处理后的叉指换能器的上表面贴合PDMS腔道进行键合处理。
在其中一些实施例中，所述PDMS腔道通过下述方法制备得到：
将PDMS注入模具中，并抽真空除以去PDMS中的气泡后加热使所述PDMS固化；将所述PDMS从模具的腔道中剥离出来，得到所述PDMS腔道。
在其中一些实施例中，在呈直线排列的所述细胞流经第二叉指换能器产生的聚焦声场，所述聚焦声场对所述细胞产生辐射力以使所述细胞产生形变的步骤中，所述细胞产生形变由高速相机或者显微镜记录。
在其中一些实施例中，在根据所述辐射力和所述形变的信息计算得到细胞的弹性模量的步骤中，具体包括下述步骤：
获取细胞形变信息；
对细胞受到的超声辐射力进行仿真；
基于仿真结果实现对细胞弹性模量的表征。
本申请目的之三，提供了一种终端，所述终端包括处理器、与所述处理器耦接的存储器，其中，
所述存储器存储有用于实现所述的细胞力学特性测量及分选方法的程序指令；
所述处理器用于执行所述存储器存储的所述程序指令以控制细胞分选。
本申请目的之四，提供了一种存储介质，存储有处理器可运行的程序指令，所述程序指令用于执行所述细胞力学特性测量及分选方法。
本申请采用上述技术方案，其有益效果如下：
本申请提供的细胞力学特性测量及分选系统、方法、终端及存储介质，待分选细胞经所述第一叉指换能器产生的驻波声场后排列成直线；呈直线排列的所述细胞流经所述第二叉指换能器产生的聚焦声场，所述聚焦声场对所述细胞产生辐射力以使所述细胞产生形变；所述细胞弹性模量计算单元根据所述辐射力和所述形变的信息计算得到细胞的弹性模量；所述细胞分选单元根据所述弹性模量触发对应的叉指换能器产生平面声场对所述细胞进行分选，本申请提供的细胞力学特性测量及分选系统、方法、终端及存储介质，通过超声非接触实 现对细胞弹性测量以及分选，减少对细胞的损伤。
附图说明
为了更清楚地说明本申请实施例的技术方案，下面将对本申请实施例或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面所描述的附图仅仅是本申请的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1为本申请实施例1提供的细胞力学特性测量及分选系统的结构示意图。
图2为本申请实施例2提供的细胞力学特性测量及分选方法的步骤流程图。
图3为本申请实施例2提供的制备叉指换能器的步骤流程图。
图4为本申请实施例2提供的制备叉指换能器的工艺示意图。
图5为本申请实施例2提供的在所述叉指换能器上形成PDMS腔道的步骤流程图。
图6为本申请实施例2提供的在驻波声场产生的超声辐射力的作用下细胞由球形被挤压为椭球形的示意图。
图7为本申请实施例3提供的终端结构示意图。
图8为本申请实施例4提供的存储介质的结构示意图。
具体实施方式
下面详细描述本申请的实施例，所述实施例的示例在附图中示出，其中自始至终相同或类似的标号表示相同或类似的元件或具有相同或类似功能的元件。下面通过参考附图描述的实施例是示例性的，旨在用于解释本申请，而不能理解为对本申请的限制。
在本申请的描述中，需要理解的是，术语“上”、“下”、“水平”、“内”、“外” 等指示的方位或位置关系为基于附图所示的方位或位置关系，仅是为了便于描述本申请和简化描述，而不是指示或暗示所指的装置或元件必须具有特定的方位、以特定的方位构造和操作，因此不能理解为对本申请的限制。
此外，术语“第一”、“第二”仅用于描述目的，而不能理解为指示或暗示相对重要性或者隐含指明所指示的技术特征的数量。由此，限定有“第一”、“第二”的特征可以明示或者隐含地包括一个或者更多个该特征。在本申请的描述中，“多个”的含义是两个或两个以上，除非另有明确具体的限定。
为了使本申请的目的、技术方案及优点更加清楚明白，以下结合附图及实施例，对本申请进行进一步详细说明。
实施例1
请参阅图1，本申请实施例1提供的一种细胞力学特性测量及分选系统的结构示意图，包括第一叉指换能器110、第二叉指换能器120、细胞弹性模量计算单元130、细胞定位单元140及细胞分选单元150。
需要说明的是：本申请实施例中给出的叉指换能器，是由在压电基底上镀叉指电极形成，在输入相应频率的正弦波信号时，会产生相应频率的表面波信号。由于具有高频率、微型化、低能量损耗的特点，可应用于对微观粒子或细胞的操控，同时，可以通过调控叉指换能器形态调控表面波声场，实现聚焦或平面声场的产生，通过聚焦声场形变细胞，测量细胞力学特性，通过力学特性测量结构选择性触发产生平面声场对细胞进行分选。
本申请提供的细胞力学特性测量及分选系统，其工作方式如下：
步骤S110：待分选细胞经所述第一叉指换能器110产生的驻波声场后排列成直线。可以理解，通过调控叉指换能器形态调控表面波声场，产生的驻波声场使得经过的待分选细胞排列成直线。
步骤S120：呈直线排列的所述细胞流经所述第二叉指换能器120产生的聚焦声场，所述聚焦声场对所述细胞产生辐射力以使所述细胞产生形变。可以理解，通过调控叉指换能器形态调控表面波声场，该叉指换能器120产生的高 强度聚焦声场会对细胞产生辐射力的作用，从而实现对细胞的挤压产生形变。
步骤S130：所述细胞弹性模量计算单元130根据所述辐射力和所述形变的信息计算得到细胞的弹性模量。
在其中一些实施例中，所述细胞弹性模量计算单元130包括：记录模块131及计算机132，所述记录模块131包括高速相机或者显微镜，所述细胞产生形变由所述记录模块记录，所述计算机132根据所述辐射力和所述形变的信息计算得到细胞的弹性模量。
步骤S140：所述细胞定位单元140可确定所述细胞的位置，并根据所述细胞的移动速度及与所述分选单元之间的距离，以设计延迟时间选择驱动所述细胞分选单元中的叉指换能器。
步骤在其中一些实施例中，所述细胞定位单元140包括光敏电极141。
步骤S150：所述细胞分选单元150根据所述弹性模量触发对应的叉指换能器产生平面声场对所述细胞进行分选。
在其中一些实施例中，所述细胞分选单元150包括阵列的叉指换能器151。
可以理解，当需要分选出软细胞时，所述细胞弹性模量计算单元130根据所述辐射力和所述形变的信息计算得到细胞的弹性模量，所述细胞分选单元150根据所述弹性模量触发平面超声换能器产生声波，该声波产生的辐射力会将软细胞推向腔道的上端；相反，硬细胞流经时，所述细胞分选单元150根据所述弹性模量不会触发对应的叉指换能器，硬细胞依旧沿着原方向流动，从而实现了依据细胞弹性的细胞分选。
本申请提供的细胞力学特性测量及分选系统，通过超声非接触实现对细胞弹性测量以及分选，减少对细胞的损伤。
实施例2
请参阅图2，为本申请实施例2提供的细胞力学特性测量及分选方法的步骤流程图，包括下述步骤S210至S250，以下详细说明各个步骤的实现方式。
步骤S210：待分选细胞经第一叉指换能器产生的驻波声场后排列成直线。
需要说明的是：本申请提供的叉指换能器的制备主要是通过在压电基底上镀入叉指电极和记录电极制作而成的，为了获得较大的机电耦合系数，选用128°YX或X切双面抛光的铌酸锂作为压电基底。在制作叉指换能器的过程中主要包括涂胶、光刻、镀膜、剥离等工艺，具体的制备方法包括：制备叉指换能器及在所述叉指换能器上形成PDMS(聚二甲基硅氧烷，(Polydimethylsiloxane))腔道。
请参阅图3，在制备叉指换能器的步骤中，具体包括下述步骤S310至S340，以下详细说明各个步骤的实现方式。
步骤S310：在压电基底的表面涂覆正光刻胶。
具体地，在完全清晰清洗干净的压电基底的表面，将正光刻胶AZ5214以3000rpm旋涂30s，将压电基底放置在65℃加热板上烘烤3min。利用台阶仪对光刻胶的厚度进行测试，光刻胶的厚度大概为1.5μm，如图4中(a)。
步骤S320：将涂覆有正光刻胶的压电基底进行曝光和显影处理。
具体地，然后将制作好的菲林片覆盖在上面图4中(a)上面进行曝光，有图案部分不透光，无图案部分透光，有光透过的部分会固化，在采用mif300进行显影的时候固化部分被溶解，非固化部分不会被溶解，显影形成如图4中(b)所示的图形。
步骤S330：对曝光和显影处理后的压电基底进行磁控溅射，使其表面生长金属电极层或光敏电极层。
具体地，对已完成图形转移的基底进行磁控溅射，使其生长厚度约为200nm的金属电极或光敏电极层，如图4中(c)。
步骤S340：将生长电极层的压电基底剥离光刻胶，完成叉指换能器的制备。
具体地，将生长电极的基底放在丙酮溶液中，利用超声清洗机的超声波震 动剥离光刻胶，完成叉指换能器的制作，如图4中(d)。
请参阅图5，在所述叉指换能器上形成PDMS腔道的步骤中，具体包括下述步骤S410至步骤S440，以下详细说明各个步骤的实现方式。
步骤S410：对所述叉指换能器进行预处理。
具体地，通过酸洗、醇洗和水洗等方法除去所述叉指换能器表面残留杂质，如灰尘和有机吸附物等，最后将所述叉指换能器置于洁净处晾干。
步骤S420：在预处理后的所述叉指换能器的表面涂覆负光刻胶并烘干。
具体地，利用涂胶机在预处理后的所述叉指换能器的表面旋凃SU-8(50)负光刻胶，其中旋凃参数如下：3000rpm，30s，SU-8(50)的厚度大约为50μm；涂胶完后，将所述叉指换能器水平放置在95℃加热板45min，让光刻胶中的溶剂挥发，以增强光刻胶与所述叉指换能器之间的黏附力，得到图4中(e)的图形。
步骤S430：对烘干后的所述叉指换能器进行曝光和显影处理。
具体地，将已经制作好图形的菲林片放置在已经旋涂好光刻胶的叉指换能器上，通过曝光机对光刻胶进行曝光，曝光剂量为200mJ/cm 2。曝光后65℃加热1min；95℃加热3-5min；用显影液浸泡曝光过的叉指换能器，未曝光区域光刻胶被溶解，曝光区域光刻胶继续保留，得到图4中(h)中图形。
步骤S440：在曝光和显影处理后的叉指换能器的上表面贴合PDMS腔道进行键合处理。
在其中一些实施例中，所述PDMS腔道通过下述方法制备得到：将PDMS注入模具中，并抽真空除以去PDMS中的气泡后加热使所述PDMS固化；将所述PDMS从模具的腔道中剥离出来，得到所述PDMS腔道。
具体地，PDMS的A胶与B胶按质量比10:1进行配比，混合均匀，到入模具中，抽真空除去PDMS中的气泡，最后将培养皿放在80℃烘箱内30min，使PDMS固化，如图4中(g)所示；将PDMS腔道从模具中剥离出来，得到 如图4中(h)所示。
进一步地，将已经制作好的叉指换能器和PDMS腔道进行等离子处理，等离子处理的功率为150W，持续时间70s，然后将PDMS腔道贴合在叉指换能器上进行键合，80℃烘箱中烘烤20min。得到图4中(I)所示制作好的用于实验的叉指换能器。
步骤S220：呈直线排列的所述细胞流经第二叉指换能器产生的聚焦声场，所述聚焦声场对所述细胞产生辐射力以使所述细胞产生形变。
在其中一些实施例中，在呈直线排列的所述细胞流经第二叉指换能器产生的聚焦声场，所述聚焦声场对所述细胞产生辐射力以使所述细胞产生形变的步骤中，所述细胞产生形变由高速相机或者显微镜记录。
步骤S230：根据所述辐射力和所述形变的信息计算得到细胞的弹性模量。
在其中一些实施例中，在根据所述辐射力和所述形变的信息计算得到细胞的弹性模量的步骤中，具体包括下述步骤S231至步骤S233，以下详细说明各个步骤的具体实现方式。
步骤S231：获取细胞形变信息。
可以理解，在细胞流经声场区域时，首先使用低强度驻波声场将细胞排列在声波节点位置，随后使用高强度聚焦的驻波超声对细胞进行作用，此时细胞膜两端受到的超声辐射力方向相反，从而对细胞形成挤压，产生形变。细胞形变量主要取决于外力和细胞弹性模量，外力是细胞形变的主要因素，与细胞形变量成正比。如图6所示，在驻波声场产生的超声辐射力的作用下细胞由球形被挤压为椭球形，基于图像处理算法可以对细胞的轮廓进行提取和分析。
步骤S232：对细胞受到的超声辐射力进行仿真。
具体地，基于已有声波能量的测量结果及细胞声速及密度等信息对细胞受到的超声辐射力进行仿真。
可以理解，为了获得细胞受到的超声辐射力，需要对细胞所处的声场信息 进行表征，然后应用声辐射应力张量理论进行计算。入射驻波声场可以表示为：
p in＝p asin(k oy)e -iωt
其中：p a为声压幅度，k o为外在介质中的波束，ω＝2πf为角频率，f为超声频率。在细胞内外的液体介质中，声压p 1的声波传播方程为亥姆霍兹方程：
Figure PCTCN2022123288-appb-000001
其中：Δ为拉普拉斯算子，c为介质的声速，超声粒子速度可以用欧拉公式表示为：
Figure PCTCN2022123288-appb-000002
其中：ρ为介质的密度。基于以上超声声场信息，可以得到超声辐射应力张量(T)：
Figure PCTCN2022123288-appb-000003
其中：i表示二维空间的平均张量，<·>代表声波周期内的时间平均。
步骤S233：基于仿真结果实现对细胞弹性模量的表征。
可以理解，在获得细胞形变和细胞受到的力学信息后，需要搭建力、细胞形变和弹性模量模型，从而计算细胞弹性模量。
为了模拟细胞膜的平面内弹性，通常采用neo-Hookean定律，实际的三维细胞在面内受到拉伸力可以用细胞的二维模型来描述。在这种情况下，由neo-Hookean定律推导出弹性模量公式为：
Figure PCTCN2022123288-appb-000004
其中E s表示细胞膜弹性模量，λ＝ds/ds 0代表细胞膜的弯曲拉伸。ds 0和ds分别表示形变前后细胞膜的无限小内联单元。
通过上述步骤S231至步骤233可以完成根据所述辐射力和所述形变的信息计算得到细胞的弹性模量。
步骤S240：根据所述细胞的移动速度及与所述分选单元之间的距离，设计延迟时间选择驱动叉指换能器。
在本实施例中，本申请通过设计细胞定位单元以确定所述细胞的位置，并根据所述细胞的移动速度及与所述分选单元之间的距离，以设计延迟时间选择驱动所述细胞分选单元中的叉指换能器。
步骤S250：所述弹性模量触发对应的叉指换能器产生平面声场对所述细胞进行分选。
可以理解，当需要分选出软细胞时，所述细胞弹性模量计算单元130根据所述辐射力和所述形变的信息计算得到细胞的弹性模量，所述细胞分选单元150根据所述弹性模量触发平面超声换能器产生声波，该声波产生的辐射力会将软细胞推向腔道的上端；相反，硬细胞流经时，所述细胞分选单元150根据所述弹性模量不会触发对应的叉指换能器，硬细胞依旧沿着原方向流动，从而实现了依据细胞弹性的细胞分选。
本申请提供的细胞力学特性测量及分选方法，通过超声非接触实现对细胞弹性测量以及分选，减少对细胞的损伤。
实施例3
请参阅图7，为本申请实施例3的终端结构示意图。该终端50包括处理器51、与处理器51耦接的存储器52。
存储器52存储有用于实现上述CT胰腺肿瘤自动分割方法的程序指令。
处理器51用于执行存储器52存储的程序指令以控制CT胰腺肿瘤自动分割。
其中，处理器51还可以称为CPU(Central Processing Unit，中央处理单元)。处理器51可能是一种集成电路芯片，具有信号的处理能力。处理器51还可以是通用处理器、数字信号处理器(DSP)、专用集成电路(ASIC)、现成可编程门阵列(FPGA)或者其他可编程逻辑器件、分立门或者晶体管逻辑器件、分立硬件组件。通用处理器可以是微处理器或者该处理器也可以是任 何常规的处理器等。
实施例4
请参阅图8，为本申请实施例4的存储介质的结构示意图。本申请实施例的存储介质存储有能够实现上述所有方法的程序文件61，其中，该程序文件61可以以软件产品的形式存储在上述存储介质中，包括若干指令用以使得一台计算机设备(可以是个人计算机，服务器，或者网络设备等)或处理器(processor)执行本申请各个实施方式方法的全部或部分步骤。而前述的存储介质包括：U盘、移动硬盘、只读存储器(ROM，Read-Only Memory)、随机存取存储器(RAM，Random Access Memory)、磁碟或者光盘等各种可以存储程序代码的介质，或者是计算机、服务器、手机、平板等终端设备。
可以理解，以上所述实施例的各技术特征可以进行任意的组合，为使描述简洁，未对上述实施例中的各个技术特征所有可能的组合都进行描述，然而，只要这些技术特征的组合不存在矛盾，都应当认为是本说明书记载的范围。
以上仅为本申请的较佳实施例而已，仅具体描述了本申请的技术原理，这些描述只是为了解释本申请的原理，不能以任何方式解释为对本申请保护范围的限制。基于此处解释，凡在本申请的精神和原则之内所作的任何修改、等同替换和改进，及本领域的技术人员不需要付出创造性的劳动即可联想到本申请的其他具体实施方式，均应包含在本申请的保护范围之内。

Claims (13)

  1. 一种细胞力学特性测量及分选系统，其特征在于，包括：第一叉指换能器、第二叉指换能器、细胞弹性模量计算单元、细胞定位单元及细胞分选单元，其中：
    待分选细胞经所述第一叉指换能器产生的驻波声场后排列成直线；呈直线排列的所述细胞流经所述第二叉指换能器产生的聚焦声场，所述聚焦声场对所述细胞产生辐射力以使所述细胞产生形变；所述细胞弹性模量计算单元根据所述辐射力和所述形变的信息计算得到细胞的弹性模量；所述细胞定位单元用于确定所述细胞的位置，所述细胞单位单元还用于根据所述细胞的移动速度及与所述分选单元之间的距离，以设计延迟时间选择驱动所述细胞分选单元中的叉指换能器；所述细胞分选单元根据所述弹性模量触发对应的叉指换能器产生平面声场对所述细胞进行分选。
  2. 如权利要求1所述的细胞力学特性测量及分选系统，其特征在于，所述细胞弹性模量计算单元包括：记录模块及计算机，所述记录模块包括高速相机或者显微镜，所述细胞产生形变由所述记录模块记录，所述计算机根据所述辐射力和所述形变的信息计算得到细胞的弹性模量。
  3. 如权利要求1所述的细胞力学特性测量及分选系统，其特征在于，所述细胞定位单元包括光敏电极。
  4. 如权利要求1所述的细胞力学特性测量及分选系统，其特征在于，所述细胞分选单元包括阵列的叉指换能器。
  5. 一种细胞力学特性测量及分选方法，其特征在于，包括下述步骤：
    待分选细胞经第一叉指换能器产生的驻波声场后排列成直线；
    呈直线排列的所述细胞流经第二叉指换能器产生的聚焦声场，所述聚焦声场对所述细胞产生辐射力以使所述细胞产生形变；
    根据所述辐射力和所述形变的信息计算得到所述细胞的弹性模量；
    根据所述细胞的移动速度及与所述分选单元之间的距离，设计延迟时间选择驱动叉指换能器；
    所述弹性模量触发对应的叉指换能器产生平面声场对所述细胞进行分选。
  6. 如权利要求5所述的细胞力学特性测量及分选方法，其特征在于，所述叉指换能器包括下述制备方法：制备叉指换能器及在所述叉指换能器上形成PDMS腔道。
  7. 如权利要求6所述的细胞力学特性测量及分选方法，其特征在于，在制备叉指换能器的步骤中，具体包括下述步骤：
    在压电基底的表面涂覆正光刻胶；
    将涂覆有正光刻胶的压电基底进行曝光和显影处理；
    对曝光和显影处理后的压电基底进行磁控溅射，使其表面生长金属电极层或光敏电极层；
    将生长电极层的压电基底剥离光刻胶，完成叉指换能器的制备。
  8. 如权利要求6所述的细胞力学特性测量及分选方法，其特征在于，在所述叉指换能器上形成PDMS腔道的步骤中，具体包括下述步骤：
    对所述叉指换能器进行预处理；
    在预处理后的所述叉指换能器的表面涂覆负光刻胶并烘干；
    对烘干后的所述叉指换能器进行曝光和显影处理；
    在曝光和显影处理后的叉指换能器的上表面贴合PDMS腔道进行键合处理。
  9. 如权利要求8所述的细胞力学特性测量及分选方法，其特征在于，所述PDMS腔道通过下述方法制备得到：
    将PDMS注入模具中，并抽真空除以去PDMS中的气泡后加热使所述PDMS固化；将所述PDMS从模具的腔道中剥离出来，得到所述PDMS腔道。
  10. 如权利要求5所述的细胞力学特性测量及分选方法，其特征在于，在呈 直线排列的所述细胞流经第二叉指换能器产生的聚焦声场，所述聚焦声场对所述细胞产生辐射力以使所述细胞产生形变的步骤中，所述细胞产生形变由高速相机或者显微镜记录。
  11. 如权利要求5所述的细胞力学特性测量及分选方法，其特征在于，在根据所述辐射力和所述形变的信息计算得到细胞的弹性模量的步骤中，具体包括下述步骤：
    获取细胞形变信息；
    对细胞受到的超声辐射力进行仿真；
    基于仿真结果实现对细胞弹性模量的表征。
  12. 一种终端，其特征在于，所述终端包括处理器、与所述处理器耦接的存储器，其中，
    所述存储器存储有用于实现权利要求4-11任一项所述的细胞力学特性测量及分选方法的程序指令；
    所述处理器用于执行所述存储器存储的所述程序指令以控制细胞分选。
  13. 一种存储介质，其特征在于，存储有处理器可运行的程序指令，所述程序指令用于执行权利要求4至11任一项所述细胞力学特性测量及分选方法。
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