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CN-115287186-B - Three-dimensional ultrasonic field device and method for performing 3D arrangement on cells

CN115287186BCN 115287186 BCN115287186 BCN 115287186BCN-115287186-B

Abstract

The invention discloses a three-dimensional ultrasonic sound field device and a method for performing cell 3D arrangement, wherein the three-dimensional ultrasonic sound field device comprises a sound field generating device, a cell culture pond, a device base and a signal generator, wherein the sound field generating device is provided with a cavity for accommodating a cell culture pond, piezoelectric ceramics are respectively arranged on the inner wall and the bottom wall of the periphery of the cavity, two poles of each piezoelectric ceramic are connected with the signal generator, the cell culture pond can be arranged in the cavity in a separated mode and is used for placing experimental samples to be arranged, the device base is used for fixing the sound field generating device, the signal generator is used for generating sine wave signals with different frequencies and amplitudes and respectively and independently applying sine wave signals with different frequencies and amplitudes to each piezoelectric ceramic, and different piezoelectric ceramics generate different interference patterns according to the received sine wave signals with different amplitudes, so that cells in the cell culture pond generate different arrangement modes. The three-dimensional ultrasonic sound field device uses multi-wave interference in the three-dimensional direction, realizes a sound field mode in the three-dimensional space, and can meet various configurations of suspended cells in the space.

Inventors

  • TIAN LIANGFEI
  • JIANG XINGYU
  • ZHAO YUNPENG

Assignees

  • 浙江大学滨江研究院
  • 杭州博日科技股份有限公司

Dates

Publication Date
20260508
Application Date
20220804

Claims (7)

  1. 1.A three-dimensional ultrasonic field device for 3D alignment of cells, comprising: the sound field generating device is provided with a cavity for accommodating the cell culture pond, piezoelectric ceramics are respectively arranged on the inner wall and the bottom wall around the cavity, the two poles of each piezoelectric ceramic are connected with the signal generator, and five piezoelectric ceramics generate coherent ultrasonic beams to generate a three-dimensional ultrasonic sound field in the cavity; The cell culture pond can be arranged in the cavity in a separating way and is used for placing experimental samples to be arranged; The device base is detachably connected with the sound field generating device and is used for fixing the sound field generating device; The signal generator is used for generating sine wave signals with different frequencies and amplitudes according to the arrangement requirement of cells, and respectively and independently applying the sine wave signals to each piezoelectric ceramic, and the different piezoelectric ceramics generate different interference patterns according to the received sine wave signals with different amplitudes so that cells in the cell culture pond generate different arrangement modes; let piezoelectric ceramics on the inner wall around the cavity be P1-P4 respectively, piezoelectric ceramics on the bottom wall of the cavity be P5, then: When the amplitude values of P1-P5 are simultaneously maximum values, the cells are arranged in a three-dimensional lattice manner in the cell culture pond; when the amplitude of P1-P4 is the maximum value and the amplitude of P5 is 0, the cells are arranged in a line shape parallel to the Z-axis direction in the cell culture pond; when the amplitude of P5 is the maximum value and the rest amplitudes are 0, the cells in the cell culture pond are in planar arrangement parallel to the xoy plane, wherein the xoy plane is parallel to the P5; When the amplitude of P1 and P2 is the maximum value and the rest amplitude is 0, the cells in the cell culture pond are arranged in a plane shape in the direction parallel to the angular bisector of the direction of the included angle formed by P1 and P2; when the amplitudes of P1 and P3 are the maximum and the rest are 0, the cells in the cell culture pond can be arranged in a plane parallel to the xoz plane, and the xoz plane is parallel to the P1 and P3 planes.
  2. 2. The three-dimensional ultrasonic sound field device for 3D arrangement of cells according to claim 1, wherein the sound field generating device and the device base are made of resin materials.
  3. 3. The three-dimensional ultrasonic field device for 3D arrangement of cells of claim 1, wherein the cell culture pond is made of transparent material.
  4. 4. A three-dimensional ultrasound field device for 3D alignment of cells according to claim 3, wherein the cell culture pond is made of quartz glass.
  5. 5. The device for three-dimensional ultrasonic field of 3D arrangement of cells according to claim 1, wherein the bottom of the chamber is provided with a limiting groove for limiting the arrangement culture pond.
  6. 6. The three-dimensional ultrasonic field device for 3D alignment of cells of claim 5, wherein the limiting groove comprises at least two L-shaped limiting brackets.
  7. 7. A method for 3D alignment of cells using the three-dimensional ultrasound field apparatus according to any of claims 1-6, comprising the steps of: (1) Placing the sound field generating device on a device base, placing the cell culture pond into a cavity of the sound field generating device, and adding purified water into the cavity; (2) Adding the cell dispersion to a cell culture pond; (3) According to the sound field pattern required by cell arrangement, corresponding amplitude modulation is carried out on corresponding piezoelectric ceramics, so that different interference sound waves are generated in the space in the cell culture pond by the generated sound waves, and the cells are subjected to 3D arrangement.

Description

Three-dimensional ultrasonic field device and method for performing 3D arrangement on cells Technical Field The invention relates to the technical field of tissue engineering, in particular to a three-dimensional ultrasonic sound field device and a method for performing 3D arrangement on cells. Background The alignment of cells and other biological samples into desired patterns plays an important role in many biological and biomedical research studies, such as cell-cell interactions, tissue engineering, and regenerative medicine. Cells are fixed in the acoustic potential well through the mutual interference of a plurality of ultrasonic waves, and the arrangement mode of the cells is changed according to the change of a sound field model, so that the method has become a relatively common cell control mode. For example, chinese patent publication No. CN110643486a discloses an ultrasonic field device and a method for manufacturing a digital PCR droplet array chip, in which a sine wave is applied to a piezoelectric ceramic device having the same resonance frequency by a signal generator, and the radiation force generated by the sound wave can capture the droplets in the emulsion, so that two-dimensional arrangement is generated at a specific position. For example, a chinese patent document with publication No. CN202010061217.9 discloses a hexagonal surface wave acoustic tweezer chip for cell arrangement and assembly, the chip comprising a hexagonal surface wave acoustic tweezer and a microfluidic cavity, both of which are connected together by plasma bonding, wherein the microfluidic cavity is configured to accommodate a cell solution, a microfluidic channel is disposed at the bottom of the microfluidic cavity, at least one cell solution sample inlet and at least one cell solution sample outlet are further disposed on the microfluidic cavity, the hexagonal surface wave acoustic tweezer is configured to generate coherent beams, and the hexagonal surface wave acoustic tweezer is disposed around the microfluidic cavity, the hexagonal surface acoustic tweezer comprises a Z-cut lithium niobate substrate, and six interdigital transducers disposed on the Z-cut lithium niobate substrate, two poles of each interdigital transducer are led out by wires, the wires are sequentially connected with a power amplifier and a signal source, when cell arrangement is required, the signal source generates radio frequency signals, the radio frequency signals are amplified to specific powers by the power amplifier and then applied to the specific interdigital transducers, the different transducers are activated or modulated in phase according to different signal configurations, each transducer generates acoustic wave beams, and the acoustic wave generated by the transducers are transmitted to the microfluidic cavity and generate interference patterns in the cell arrangement pattern, and the acoustic wave patterns are subjected to cell arrangement, thereby the cell arrangement is realized. The traditional acoustic tweezers for cell manipulation mostly adopt a surface wave mode, only can control cells on a substrate in a limited way, two pairs of piezoelectric ceramics which are perpendicular to each other are used for generating a standing wave field, and the mode of forming a tetragonal lattice sound field through standing wave length is used for controlling cells or other particles, which are limited on the surface of the substrate, and the method does not involve arranging and assembling the cells in a three-dimensional space, so that the cells are used for a certain configuration in the dimension of the cell space, and the requirement of arranging suspended cells in the three-dimensional space is difficult to meet. Disclosure of Invention Aiming at the defect that the existing ultrasonic sound field device cannot realize the arrangement of cells in a three-dimensional space, the invention provides a three-dimensional ultrasonic sound field device for 3D arrangement of cells, which not only can enable the cells to be arranged in the space, but also can enable the cells to have different arrangement patterns by modulating amplitude values of different piezoelectric ceramics. The technical scheme of the invention is as follows: A three-dimensional ultrasonic field device for 3D alignment of cells, comprising: the sound field generating device is provided with a cavity for accommodating the cell culture pond, piezoelectric ceramics are respectively arranged on the inner wall and the bottom wall around the cavity, the two poles of each piezoelectric ceramic are connected with the signal generator, and five piezoelectric ceramics generate coherent ultrasonic beams to generate a three-dimensional ultrasonic sound field in the cavity; The cell culture pond can be arranged in the cavity in a separating way and is used for placing experimental samples to be arranged; The device base is detachably connected with the sound field generating dev