阅读说明：本技术 法拉第波多波长合成的细胞组装方法及应用 (Cell assembly method for Faraday wave multi-wavelength synthesis and application ) 是由 陈璞 谷龙军 于 2021-07-30 设计创作，主要内容包括：本发明公开了一种法拉第波多波长合成的生物组装方法及应用,本方法具有系统易搭建、操控简单、图案动态可调以及生物相容性良好等优势。本方法区别于现有声学生物组装方法单一波长条件下的细胞操控原理,通过把不同波长的正弦或余弦信号进行合成,将传统法拉第波频域下单一波长组装模式提升为多波长组装模式,实现液底多尺度细胞复杂、任意图案排列,使其更加适合组织工程及生物制造中对于细胞复杂排列的需求,具有巨大的应用前景和商业化价值。(The invention discloses a biological assembly method for Faraday wave multi-wavelength synthesis and application thereof. The method is different from the cell control principle under the condition of single wavelength in the existing acoustic biological assembly method, and by synthesizing sine or cosine signals with different wavelengths, the single-wavelength assembly mode under the traditional Faraday wave frequency domain is improved into a multi-wavelength assembly mode, so that the complex and random pattern arrangement of multi-scale cells at the liquid bottom is realized, the method is more suitable for the requirement of complex arrangement of cells in tissue engineering and biological manufacturing, and has huge application prospect and commercial value.)

1. A biological assembly method for Faraday wave multi-wavelength synthesis is characterized in that: based on the fact that any complex periodic pattern can be simplified into a series of sine waves or cosine waves by utilizing Fourier series, the method synthesizes sine or cosine signals with different wavelengths, further excites and forms Faraday waves synthesized by multiple wavelengths, and finally realizes the complex and random arrangement of multi-scale cells; the method comprises the following steps:

s1: according to the biological assembly pattern to be constructed, synthesizing sine or cosine signals with different wavelengths in waveform creating and editing tool software, and introducing a synthesized multi-wavelength signal file into an arbitrary waveform/function signal generator;

s2: opening the multi-wavelength signal file synthesized in the step S1 in the arbitrary waveform/function signal generator, outputting a corresponding electric signal through the arbitrary waveform/function signal generator, transferring the electric signal to a power amplifier for power amplification, transferring the amplified fidelity electric signal to a vibration exciter for generating stable and periodic vibration, connecting the assembly chamber with the vibration exciter and carrying out horizontal calibration;

s3: and uniformly adding the cell assembly unit-containing suspension to be assembled into an assembly chamber, and performing Faraday wave multi-wavelength synthesis assembly after the cell assembly unit-containing suspension is settled to the bottom of the assembly chamber.

2. A method of bioassembly of faraday wave multi-wavelength synthesis according to claim 1, characterized in that:

in step S1, the synthesis of the multiple sine or cosine signals specifically includes:

for the synthesis of a two-wavelength faraday wave, the function of the sinusoidal signal written is y ═ a₁*sin(f₁*2*π*x)+A₂*sin(f₂2 x pi x); wherein f is₁At a low driving frequency, f₂At a high driving frequency, A₁Amplitude, A, corresponding to a function of the sinusoidal signal at low drive frequency₂Corresponding amplitude values for the high drive frequency sinusoidal signal function; the function of the written cosine signal is y ═ A₁*cos(f₁*2*π*x)+A₂*cos(f₂2 x pi x); wherein f is₁At a low driving frequency, f₂At a high driving frequency, A₁Amplitude, A, corresponding to a function of the sinusoidal signal at low drive frequency₂Corresponding amplitude values for the high drive frequency sinusoidal signal function; for the synthesis of a multi-wavelength faraday wave, the function of the sinusoidal signal written is y ═ a₁*sin(f₁*2*π*x)+A₂*sin(f₂*2*π*x)+A₃*sin(f₃*2*π*x)+A₄*sin(f₄*2*π*x)+A₅*sin(f₅2 x pi x) + …; the function of the written cosine signal is y ═ A₁*cos(f₁*2*π*x)+A₂*cos(f₂*2*π*x)+A₃*cos(f₃*2*π*x)+A₄*cos(T₄*2*π*x)+A₅*cos(f₅*2*π*x)+...。

3. A method of bioassembly of faraday wave multi-wavelength synthesis according to claim 2, characterized in that: the driving frequency is selected to be between 1-1000 Hz.

4. A method of cell assembly by Faraday wave multi-wavelength synthesis according to claim 3, whereinThe method comprises the following steps: in the synthesis of the dual-wavelength Faraday wave, the low-driving-frequency sine or cosine signal function corresponds to the amplitude A₁Amplitude A corresponding to sine or cosine signal function with high driving frequency₂The ratio range of (A) to (B) is 1:1-1: 5; in the synthesis of multi-wavelength Faraday waves, the drive frequency f₁＜f₂＜f₃＜f₄＜f₅Amplitude A₁≤A₂≤A₃≤A₄≤A₅The ratio of the lower amplitude to the higher amplitude is in the range of 1:1 to 1: 5.

5. A method of cell assembly according to any one of claims 1 to 4 for Faraday wave multi-wavelength synthesis, wherein:

in step S2, the shape of the assembly chamber is: any one of a circle, a square, a rectangle, a triangle, a trapezoid, a diamond, a hexagon or an octagon, wherein the diameter of an outer circle of each shape assembly chamber is 0.5-20 cm; the height is 0.2-10 mm.

6. A Faraday wave multi-wavelength synthesized cell assembly method according to claim 5, wherein:

in the step S3, the cell-containing assembly unit is one or a mixture of several of a single cell, a micro-tissue block, an organoid, a cell microsphere, a cell-containing hydrogel microsphere and a cell-containing carrier particle, and the diameter is 2 to 5000 μm.

7. A method of cell assembly by Faraday wave multi-wavelength synthesis, according to claim 6, wherein:

in the step S3, the number of the cell-containing assembly units is 2-10¹²And (4) respectively.

8. A method of cell assembly by Faraday wave multi-wavelength synthesis, according to claim 7, wherein: in the step S3, the selected cells are any one or a mixture of embryonic stem cells, induced pluripotent stem cells, cancer stem cells and mesenchymal stem cells; or selecting cells from brain, liver, kidney, pancreas, blood vessel, heart, skin, bone marrow, bone, cartilage and muscle tissue organ primary cells, progenitor cells, precursor cells and disease cells.

9. A method of cell assembly by Faraday wave multi-wavelength synthesis, according to claim 8, wherein:

in the step S3, the suspension system is any one of a phosphate buffer, a cell culture medium, a natural hydrogel, a synthetic hydrogel or a mixed hydrogel.

10. Use of a method of cell assembly according to faradaic multi-wavelength synthesis according to any one of claims 1 to 4 or 6 to 9, wherein: the method is applied to the construction of physiological and pathological models of tissues and organs, the research of physiological and pathological mechanisms, the drug screening and the tissue repair.

Technical Field

The invention belongs to the technical field of tissue engineering and biological manufacturing under biomedical engineering, and particularly relates to a biological assembly method for Faraday wave multi-wavelength synthesis and application thereof.

Background

In human tissue organs, cells are interconnected by self-organization and form highly ordered cellular structures. The arrangement mode of cells in the in-vitro reduction structure and the geometric shape of the structure play a key role in simulating the shape of in-vivo tissues and organs on a macroscopic scale and play an important role in regulating the microenvironment of the cells.

In recent years, bio-fabrication has shown great potential in constructing cellular microstructures that are highly reducing the complexity and functionality of human tissue organs. Biomanufacturing is the construction of living cells, biological materials and stimulating factors into biological products with specific structures and functions using bioprinting or bio-assembly followed by tissue maturation processes. Among them, bio-assembly is considered as an important construction strategy for bio-fabrication, which has advantages in that bio-assembly can have better biocompatibility and higher efficiency in constructing cell structures than bio-printing. To date, a series of bio-assembly techniques have utilized the interaction of an external physical field with cells to manipulate the arrangement of cells in space. These physical fields include gravitational, magnetic, and electric fields, among others. However, cell structures constructed by the bio-assembly technology based on the above principle often have only simple geometric shapes such as spherical, strip-shaped, and ring-shaped shapes, and it is difficult to satisfy the requirement of constructing multi-scale, complex, and arbitrary cell structures.

Compared with other biological assembly technologies, the acoustic biological assembly has the unique advantage that the sound field can be dynamically adjusted, so that better diversity of cell structures can be provided. The acoustic holography technology is used for constructing a complex and arbitrary cell structure, but the technology has the problems of complex system, complex operation flow, incapability of realizing real-time adjustment on the complex cell structure and the like. Compared with the technology, the Faraday wave biological assembly technology has the advantages of easy system construction, simple control, dynamically adjustable patterns, good biocompatibility and the like, and has wide application prospect in the field of biological tissue engineering. However, in the patent number issued for public invention (US20160145566a1, WO2016069493a2), the original faraday wave biological assembly technology only performs single-wavelength cell assembly in the frequency domain, and the cell structure limited in a single scale can be constructed, which fails to meet the requirement for constructing multi-scale, complex and arbitrary cell structures.

Therefore, the biological assembly method capable of meeting the requirement of realizing complex and random arrangement of multi-scale cells in biological tissue engineering research has important application prospect and technical popularization and application value.

Disclosure of Invention

The invention aims to solve the problem of providing a cell assembly method for Faraday wave multi-wavelength synthesis and application aiming at the defects in the existing biological assembly technology. The method aims to synthesize a plurality of sine or cosine signals with different wavelengths, further excite and form the multi-wavelength synthesized Faraday wave, expand a single-wavelength assembly mode under the frequency domain of the traditional Faraday wave into a multi-wavelength assembly mode, realize the complex and random pattern arrangement of liquid-bottom multi-scale cells, and further enable the Faraday wave biological assembly to have more technical popularization and application values.

To achieve the above object, the working principle of the present invention is as follows:

establishing the acoustic biological assembly theory of multi-wavelength synthesis. The cells at the liquid bottom are subjected to the combined action of acoustic pressure, buoyancy and gravity in a Faraday wave sound field, wherein the acoustic pressure plays a decisive role in the balance position of the cells, and the cells are finally balanced to the position with the lowest potential energy in the sound pressure field, namely the potential well. Any complex acoustic pressure field can be reduced to a superposition of a series of sine or cosine acoustic pressure fields using a fourier series. Therefore, a complex waveform can be formed by synthesizing sine or cosine signals with a plurality of different wavelengths, so that the requirement of complex and arbitrary arrangement of multi-scale cells is met. The multi-wavelength synthesis multi-scale complex structure is shown in fig. 1. For example, the liver is an important organ with a unique multi-scale structure, the liver of an adult is composed of 50-100 ten thousand hexagonal hepatic lobules, and the periodic hepatic lobule structural units and the multi-scale micro-physiological structure of the liver tissue are shown in fig. 2. Thus, the complex liver tissue structure can be simplified to a spatially repetitive hepatic lobular structure. Through multi-wavelength synthesis, complex acoustic pressure field distribution is realized in space, and then the construction of a multi-scale liver cell structure is realized. In addition, the human body also contains a large number of tissues and organs containing specific multi-scale structural units, for example, alveoli are hemispherical vesicles mainly composed of a monolayer of epithelial cells; the renal tubules are mainly self-organized by renal tubular epithelial cells to form hollow tubular structures. The specific tissue organ physiological structures can be constructed in vitro by Faraday wave multi-wavelength synthesis and assembly.

In order to achieve the purpose, the technical scheme of the invention is as follows:

in a first aspect, the present invention provides a cell assembly method for faraday wave multi-wavelength synthesis, comprising the steps of:

s1: according to the cell pattern of the target assembly arrangement, the synthesis of Faraday waves with different wavelengths is realized in waveform creation and editing tool software, and the frequency is selected to be between 1 Hz and 1000 Hz. E.g. a synthesis of a two-wavelength faraday wave, the function of the written sinusoidal signal being y ═ a₁*sin(f₁*2*π*x)+A₂*sin(f₂2 x pi x); the function of the written cosine signal is y ═ A₁*cos(f₁*2*π*x)+A₂*cos(f₂2 x pi x). Wherein f is₁At a low driving frequency, f₂For high driving frequency, low driving frequency sine signal function corresponds to amplitude A₁Amplitude A corresponding to high driving frequency sinusoidal signal function₂In the range of 1:1 to 1: 5. For the synthesis of a multi-wavelength faraday wave, the function of the sinusoidal signal written is y ═ a₁*sin(f₁*2*π*x)+A₂*sin(f₂*2*π*x)+A₃*sin(f₃*2*π*x)+A₄*sin(f₄*2*π*x)+A₅*sin(f₅2 x pi x) + …; the function of the written cosine signal is y ═ A₁*cos(f₁*2*π*x)+A₂*cos(f₂*2*π*x)+A₃*cos(f₃*2*π*x)+A₄*cos(f₄*2*π*x)+A₅*cos(f₅2 x pi x) + …. Wherein, the frequency f₁＜f₂＜f₃＜f₄＜f₅(ii) a Amplitude A₁≤A₂≤A₃≤A₄≤A₅The ratio of the lower amplitude to the higher amplitude is in the range of 1:1 to 1: 5. The synthesized multi-wavelength signal file is directed to an arbitrary waveform/function signal generator.

S2: the cell-containing assembly unit is used for assembly, and the selected cells are one or a mixture of more of embryonic stem cells, induced pluripotent stem cells, cancer stem cells and mesenchymal stem cells; alternatively, the cells are selected from brain, liver, kidney, pancreas, blood vessel, heart, skin, bone marrow, bone, cartilage and muscleThe tissue organ primary cells, progenitor cells, precursor cells and disease cells thereof. The cell assembly unit is one or more of single cell, micro tissue block, organoid, cell microsphere, hydrogel microsphere containing cell and carrier particle containing cell, and has diameter of 2-5000 μm. The number of cell-containing assembly units is 2-10¹²And (4) respectively. The cell-containing assembly units are uniformly dispersed in phosphate buffer, cell culture medium, natural hydrogel, synthetic hydrogel and mixed hydrogel for standby.

S3: opening the synthesized multi-wavelength signal file, outputting a specific electric signal through an arbitrary waveform/function signal generator, transferring the electric signal to a power amplifier for power amplification, and transferring the amplified fidelity electric signal to a vibration exciter to generate stable and periodic vibration. And connecting the assembly chamber with the vibration exciter and carrying out horizontal calibration. The assembly chamber can be round, square, rectangular, triangular, trapezoidal, rhombic, hexagonal or octagonal; the diameter of the circumscribed circle of each shape assembly cavity is 0.5-20 cm; the height is 0.2-10 mm.

S4: and uniformly adding the cell assembly unit-containing suspension to be assembled into an assembly chamber, and performing Faraday wave multi-wavelength synthesis assembly after the cell assembly unit-containing suspension is settled to the bottom of the assembly chamber.

In a second aspect, the present invention provides an application of the above method for cell assembly by faraday wave multi-wavelength synthesis, wherein the method comprises: the method is applied to the construction of physiological and pathological models of tissues and organs, the research of physiological and pathological mechanisms, the drug screening and the tissue repair.

The advantages and the beneficial effects of the invention are as follows:

the cell assembly method for Faraday wave multi-wavelength synthesis has the advantages of easy system construction, simple operation and control, dynamically adjustable patterns, good biocompatibility and the like. The method is different from the cell control principle under the condition of single wavelength in the existing acoustic biological assembly method, synthesizes sine or cosine signals with a plurality of different wavelengths, further excites and forms a multi-wavelength synthesized Faraday wave, expands the single-wavelength assembly mode under the frequency domain of the traditional Faraday wave into a multi-wavelength assembly mode, realizes the complex and random pattern arrangement of multi-scale cells on the liquid bottom, makes the cell complex arrangement requirements in tissue engineering and biological manufacturing more suitable, and has huge application prospect and commercial transformation potential.

Drawings

FIG. 1 is a schematic diagram of a multi-wavelength synthesis multi-scale complex structure;

FIG. 2 is a schematic diagram of a periodic structural unit of liver tissue and a multi-scale micro-physiological structure;

FIG. 3 is a schematic diagram of a single-wavelength, dual-wavelength, and three-wavelength polystyrene microsphere pattern obtained by liquid-bottom assembly in example 1 of the present invention;

FIG. 4 is a schematic diagram of the microsphere pattern of single-wavelength and dual-wavelength HepG2 cells obtained by liquid-bottom assembly in example 2 of the present invention.

Detailed Description

The scheme of the invention will be explained with reference to the examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples, where specific techniques or conditions are not indicated, are to be construed according to the techniques or conditions described in the literature in the art or according to the product specifications.

Example 1: arrangement of Faraday wave multi-wavelength synthesis assembly for polystyrene microspheres

Polystyrene microspheres were added to 0.01M phosphate buffer containing 0.05% tween 20 and dispersed thoroughly. The polystyrene microspheres are red in color, 100 mu m in diameter and 1.05g/cm in density³The physical properties are similar to those of the assembled unit containing cells, and the concentration of the polystyrene microspheres is 30 mg/mL. In this example, polystyrene microspheres were used to mimic cell-containing assembled units.

1. The polystyrene microsphere is arranged under the condition of single wavelength, and the method comprises the following steps:

after the waveform is set to be a continuous sine wave, the driving frequency is set to be 20Hz, 28Hz and 58Hz, the signal amplitude is set to be 60-100mVpp, a specific electric signal is output by adjusting an arbitrary waveform/function signal generator (Tecktronix, AFG3052C), the electric signal is transferred to a power amplifier (DAYTONAUDIO, DTA-120) through a BNC female-to-RCA male line for power amplification, and the amplified fidelity electric signal is transferred to a vibration exciter (Seisano-tin-Aao science and technology Limited, SA-JZ005T) through an RCA revolution four-core socket to generate stable and periodic vibration. After a circular assembly chamber (Φ ═ 3cm) was rigidly connected to the exciter by M5 × 30, a 360 ° horizontal calibration was performed by a bubble disk level. And finally, uniformly adding the polystyrene particle suspension to be assembled into the circular assembly chamber, and performing Faraday wave assembly after the polystyrene particles are settled to the bottom of the assembly chamber. The pattern obtained by assembling the polystyrene microspheres under the condition of single wavelength is only capable of forming a relatively simple divergent, petal-shaped and concentric circular ring pattern as shown in a in fig. 3.

2. The arrangement of polystyrene microspheres under the condition of dual-wavelength synthesis comprises the following steps:

the synthesis of two sinusoidal signals of different wavelengths is realized by means of the software ArbExpress. The wave function of the synthetic sinusoidal signal at the drive frequencies of 20Hz and 28Hz corresponding wavelengths is y-1 h5 x sin (20 x 2 x pi x) + sin (28 x 2 pi x); the wave function of the synthetic sinusoidal signal at the drive frequencies of 20Hz and 58Hz corresponding wavelengths is y 1.2 sin (20 x 2 pi x) + sin (58 x 2 pi x); the wave function of the resulting sinusoidal signal at the drive frequencies of 28Hz and 28Hz corresponding wavelengths is y-sin (20 x 2 x pi x) + sin (28 x 2 x pi x). The synthesized two-wavelength sinusoidal signal is saved as a "TFW" format file and introduced into an arbitrary waveform/function signal generator (Tecktronix, AFG 3052C).

After the waveform is set to be an arbitrary waveform, a synthesized dual-wavelength sinusoidal signal file is sequentially opened, the signal amplitude is 140-160mVpp, a specific electric signal is output through an arbitrary waveform/function signal generator, the electric signal is transferred to a power amplifier (DAYTONAUDIO, DTA-120) through a BNC female-to-RCA male line for power amplification, and the amplified fidelity electric signal is transferred to a vibration exciter (SA-JZ 005T, Wuxi Aoshi technology Co., Ltd.) through an RCA revolution four-core socket for generating stable and periodic vibration. After a circular assembly chamber (Φ ═ 3cm) was rigidly connected to the exciter by M5 × 30, a 360 ° horizontal calibration was performed by a bubble disk level. And finally, uniformly adding the polystyrene particle suspension to be assembled into the circular assembly chamber, and performing Faraday wave assembly after the polystyrene particles are settled to the bottom of the assembly chamber. The pattern obtained by assembling the polystyrene microspheres under the condition of double wavelengths is shown as b in fig. 3, so that the corresponding patterns under the condition of single wavelength are superposed to obtain the patterns of the divergent shape, the petal shape and the concentric circular ring shape which are mutually superposed in pairs.

3. The polystyrene microsphere is arranged under the condition of three-wavelength synthesis, and the steps are as follows:

the synthesis of three sinusoidal signals of different wavelengths is realized by means of the software ArbExpress. The drive frequency was 20Hz and the wave function of the resulting sinusoidal signal at wavelengths corresponding to 28Hz and 58Hz was y 1.4 x sin (20 x 2 x pi x) + sin (28 x 2 pi x) + sin (58 x 2 pi x). The synthesized three-wavelength sinusoidal signal is saved as a "TFW" format file and introduced into an arbitrary waveform/function signal generator (Tecktronix, AFG 3052C).

After the waveform is set to be any waveform, a synthesized three-wavelength sinusoidal signal file is opened, the signal amplitude is 200mVpp, a specific electric signal is output through an arbitrary waveform/function signal generator, the electric signal is transferred to a power amplifier (DAYTONAUDIO, DTA-120) for power amplification through a BNC female-to-RCA male line, and the amplified fidelity electric signal is transferred to a vibration exciter (Seisaless Aoch technology Co., Ltd., SA-JZ005T) through an RCA revolution four-core socket to generate stable and periodic vibration. After a circular assembly chamber (Φ ═ 3cm) was rigidly connected to the exciter by M5 × 30, a 360 ° horizontal calibration was performed by a bubble disk level. And finally, uniformly adding the polystyrene particle suspension to be assembled into the circular assembly chamber, and performing Faraday wave assembly after the polystyrene particles are settled to the bottom of the assembly chamber. The pattern obtained by assembling the polystyrene microspheres under the condition of three wavelengths is shown as c in fig. 3, so that the complicated and random pattern arrangement of the polystyrene particles under the condition of multiple scales is further realized, and the assembled pattern has the characteristics of divergent patterns, petal-shaped patterns and concentric circular patterns.

Example 2: faraday wave multi-wavelength synthesis assembly for arranging HepG2 cell microspheres

The HepG2 cell microspheres are arranged under the condition of single wavelength, and the steps are as follows:

HepG2 cells were added to a six-well low-adhesion plate (Corning, 34)71) 2 x 10 per well⁶Cells were plated in 2mL DMEM complete medium. The six well low adhesion plates were placed on a plate shaker at 75rpm and cultured for three days to pellet. One well of HepG2 cell microspheres was collected and added to 0.01M phosphate buffer for use.

After the waveform is set to be a continuous sine wave, the driving frequency is set to be 42Hz and 45Hz, the signal amplitude is set to be 60-80mVpp, a specific electric signal is output by adjusting an arbitrary waveform/function signal generator (Tecktronix, AFG3052C), the electric signal is transferred to a power amplifier (DAYTONAUDIO, DTA-120) through a BNC female-to-RCA male line for power amplification, and the amplified fidelity electric signal is transferred to a vibration exciter (005T) through an RCA revolution four-core socket to generate stable and periodic vibration. After a black background circular assembly chamber (Φ ═ 2cm) was rigidly connected to the exciter via M5 × 30, a 360 ° horizontal calibration was performed by means of a bubble disk level. And finally, uniformly adding the HepG2 cell microsphere suspension to be assembled into the circular assembly chamber, and performing Faraday wave assembly after the HepG2 cell microspheres are settled to the bottom of the assembly chamber. The pattern of the assembled HepG2 cell microspheres under single wavelength conditions is shown as a in FIG. 4, and only a simple divergent, four-lobed petaloid pattern can be formed.

The arrangement of HepG2 cell microspheres under the condition of double wavelength comprises the following steps:

the synthesis of two sinusoidal signals of different wavelengths is realized by means of the software ArbExpress. The wave function of the resulting sinusoidal signal at the respective wavelengths at the drive frequencies 42Hz and 45Hz is y 2.1 sin (42 x 2 pi x) + sin (45 x 2 pi x). The synthesized two-wavelength sinusoidal signal is saved as a "TFW" format file and introduced into an arbitrary waveform/function signal generator (Tecktronix, AFG 3052C).

After the waveform is set to be any waveform, a synthesized dual-wavelength sinusoidal signal file is opened, the signal amplitude is 140-160mVpp, HepG2 cell microsphere assembly under the dual-wavelength condition is completed through the Faraday wave acoustic biological assembly system, the pattern obtained by HepG2 cell microsphere assembly under the dual-wavelength condition is shown as b in figure 4, the corresponding patterns under the single-wavelength condition are overlapped, the HepG2 cell microspheres are arranged in a complex and random pattern under the multi-scale condition, and the assembled pattern has the characteristics of a divergent and a four-leaf petal-shaped pattern.

In summary, the invention innovatively synthesizes sine or cosine signals with different wavelengths, further excites and forms a multi-wavelength synthesized Faraday wave, and realizes a complex and arbitrary cell arrangement pattern under a multi-scale condition. The method has the advantages of easy system construction, simple operation and control, dynamically adjustable pattern and better biocompatibility. More importantly, the method is different from the cell control principle under the condition of single wavelength in the existing acoustic biological assembly method, the Faraday wave synthesized by multiple wavelengths is formed by excitation, the single wavelength assembly mode under the frequency domain of the traditional Faraday wave is expanded into the multi-wavelength assembly mode, the complex and random pattern arrangement of the multi-scale cells at the liquid bottom is realized, and an innovative solution is provided for the requirements of the complex and random arrangement of the multi-scale cells in tissue engineering and biological manufacturing.