CN212528701U

CN212528701U - 3D printing device

Info

Publication number: CN212528701U
Application number: CN202021284913.8U
Authority: CN
Inventors: 林雪霞; 方飞祥
Original assignee: Huaqiao University
Current assignee: Huaqiao University
Priority date: 2020-07-03
Filing date: 2020-07-03
Publication date: 2021-02-12
Anticipated expiration: 2030-07-03

Abstract

The utility model discloses a 3D printing device, which relates to the field of 3D printing, and comprises a three-dimensional mobile platform, wherein a three-axis mechanical arm is arranged on the three-dimensional mobile platform, the three-axis mechanical arm is electrically connected with a controller, a printing nozzle is arranged on the three-axis mechanical arm, the printing nozzle is electrically connected with a driving power supply, the driving power supply is wirelessly connected with a computer, and control software is arranged in the computer; and a receiving platform is fixedly arranged on the three-dimensional moving platform, the receiving platform is positioned below the printing nozzle, and printing ink is filled in the printing nozzle. The utility model discloses utilize piezoelectric type inkjet printing technique to print the liquid drop on receiving platform to realize the accurate control of liquid drop and the device of accurate positioning. The invention has the advantages of simple operation, high printing precision, good repeatability and small pollution probability to the sample in the printing process, and effectively solves the problems of complicated structure, single printing mode and the like of the printing device.

Description

3D printing device

Technical Field

The utility model relates to a 3D prints technical field, especially relates to a 3D printing device.

Background

The 3D printing technology appeared in the last 90 th century and belongs to one of the rapid prototyping technologies. The printer has basically the same working principle as common printers, and the ink bag of the printer is filled with liquid or solid powder and other 'ink' obtained through special treatment. The pattern model which is overlapped layer by layer is printed on the basis of the digital model file through being connected with a computer, so that the composition is converted into a real object. This technology is now used in a number of areas such as aerospace, construction modeling, mold manufacturing, biomedical, garment design, and the like. Compared with the traditional technology, the most prominent advantage of the 3D printing technology is that the printing of any shape can be directly realized according to the model diagram in the computer without using mechanical processing or a mold. This saves time and money greatly, greatly improves the productivity of enterprises and reduces the production cost, fundamentally reduces the waste of materials and saves labor force.

According to the forming mode of the printing material, the 3D printing technology can be divided into three types, namely extrusion forming, powder material forming and photopolymerization forming, and each type also comprises one or more technical routes. A general printing apparatus mainly uses a single-extrusion type or a double-extrusion type printing mode, and is less likely to select the printing mode, and the printing apparatus includes a complicated structure. If layer-by-layer printing of various materials is to be realized, a plurality of extruders are required to be matched together, the printing difficulty is high, the printing efficiency is low, and a complex printing program needs to be designed. Furthermore, this common single printing mode is not satisfactory for printing by most devices.

SUMMERY OF THE UTILITY MODEL

The utility model aims at providing a 3D printing device to solve the problem that above-mentioned prior art exists, make the printing mode diversified, improved printing efficiency.

In order to achieve the above object, the utility model provides a following scheme:

the utility model provides a 3D printing device, including three-dimensional moving platform, install the triaxial arm on the three-dimensional moving platform, triaxial arm electricity is connected with the controller, install the printing shower nozzle on the triaxial arm, printing shower nozzle electricity is connected with drive power supply, drive power supply wireless connection has the computer, the computer embeds has control software; and a receiving platform is fixedly arranged on the three-dimensional moving platform, the receiving platform is positioned below the printing nozzle, and printing ink is filled in the printing nozzle.

Optionally, a plurality of printing channels are uniformly arranged in the printing nozzle, an ink bag is arranged in each printing channel, and the printing ink is contained in the ink bag; the bottom of the ink bag is provided with a nozzle, the side wall of the ink bag is provided with a piezoelectric ceramic piece, and the printing channel is electrically connected with the driving power supply through a wire; and a diaphragm is arranged on the outer side of the piezoelectric ceramic piece.

Optionally, the triaxial arm include the level fixed set up in Y axle on the three-dimensional moving platform, the slip is provided with the Z axle of vertical installation on the Y axle, Z axle top fixed mounting has the X axle that the level set up, just the horizontal projection of X axle with the horizontal projection of Y axle sets up perpendicularly, be provided with the control end through sprocket structure horizontal slip on the X axle, be provided with the arm through sprocket structure vertical slip on the control end lateral wall, the terminal fixed mounting of arm has the manipulator, print the shower nozzle install in on the manipulator.

Optionally, a connection port is formed on the driving power supply, and the connection port is connected with the printing channel through a wire.

Optionally, five printing channels are uniformly arranged in the printing nozzle, each printing channel is internally provided with an ink bag, and the five ink bags are respectively filled with different printing inks by using 5mL injectors.

Optionally, the driving power supply adopts a parallel circuit, the parallel circuit adopts a control signal to drive the two high-voltage switches, and the high-voltage signal is cut into square waves, so that the adjustment and conversion of the magnitude of the driving voltage are realized; the parallel circuit is provided with a plurality of switches in parallel, and each switch is connected with one printing channel through a lead.

The utility model also provides a 3D printing method, including the method of printing and cultivateing the cell, the step is as follows:

step one; opening the control software of the computer, inputting control parameters, and inputting the coordinates of the printing pattern model in the controller;

step two; cleaning the printing channel; injecting the cell printing suspension into the ink bag, sealing an inlet of the ink bag, and spraying the cell printing suspension onto the glass substrate by using a voltage-driven ink-jet printing head; the cell printing suspension comprises a DMEM complete culture medium, a sodium alginate aqueous solution with the mass percentage content of 0.2% -1.2% and cells to be printed; the volume ratio of the DMEM complete culture medium to the sodium alginate aqueous solution with the mass percentage content of 0.2% -1.2% is 1: 1; the concentration of the cells to be printed in the cell printing suspension was 1X 10⁶～9×10⁶Per mL; covering a polydimethylsiloxane channel on the glass substrate sprayed with the cell printing suspension, and sequentially introducing 0.2-0.3M CaCl from the sample inlet hole by using a micro-injection pump₂Aqueous solution and DMEM complete culture medium, at 35-40 deg.C and 15% -25% CO₂Culturing under the conditions of (1);

step three; starting printing, wherein the controller drives the three-axis mechanical arm to move so as to drive the printing nozzle to move according to a preset pattern coordinate; the driving power supply applies pulse waveform, the piezoelectric ceramic piece generates deformation in the rising and continuous processes of pulse voltage, and printing ink liquid drops in the ink bag are ejected out of the nozzle by utilizing the telescopic deformation of the piezoelectric ceramic piece for printing; the disconnection of the printing channel voltage is controlled and switched by control software, the working voltage during printing is set to be 50-500V, the printing interval is 100-300 mu m, and the number of printing drops is 1-9 drops; until printing is completed. The number of cells in the obtained single gel is less than or equal to 5, which is beneficial to single cell analysis.

Optionally, mixing the glass and trichlorosilane, vacuumizing to-0.8 MPa, and keeping the vacuum state for 0.8-1.2 h under the condition of-0.8 MPa to obtain a glass substrate; the height of the polydimethylsiloxane channel is 20-100 mu m, and the width of the polydimethylsiloxane channel is 0.6-1.2 mm.

The utility model discloses for prior art gain following technological effect:

the utility model discloses print the liquid drop on receiving platform to realize the accurate control and the accurate positioning of liquid drop. According to the cooperation of each element, the patterning printing operation can be efficiently carried out, and the problems of complicated structure, single printing mode and the like of the printing device are effectively solved. In addition, the ink-jet printing technology has high liquid drop generation rate, is suitable for large-scale production of liquid drops, meets the requirements of more high efficiency, high sensitivity and rapidness of sample analysis, and has great potential advantages in various fields. The application range is wide, and the method can be used for research of printing materials or single liquid drops and construction of tissues and organs in tissue engineering.

The utility model discloses use the mixed solution of sodium alginate, DMEM complete medium and cell as print medium, with cell separation, fix on culture apparatus to make the cell continuously divide division to grow in culture apparatus. By adjusting parameters such as printing voltage, drop number and the like, the method can be used for single cell separation research and can realize control of different cell numbers in a single gel. The method of the utility model has the advantages of simple operation, lower cost, high repeatability, good separation effect, good biocompatibility of the used materials, and easy preparation and operation of the culture device. The device and the method of the utility model can realize the long-term culture of cells, and have important significance for the observation and research of single cells and a plurality of cells.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without inventive labor.

Fig. 1 is a schematic structural view of the 3D printing apparatus of the present invention;

fig. 2 is a schematic view of the working state of the ink bag of the 3D printing apparatus of the present invention;

fig. 3 is a schematic view of a three-axis mechanical arm structure of the 3D printing apparatus of the present invention;

description of reference numerals: 1. a computer; 11. control software; 2. a drive power supply; 21. a connection port; 3. a three-dimensional mobile platform; 31. a controller; 32. a three-axis mechanical arm; 4. printing a spray head; 41. an ink bag; 5. a receiving platform; 6. piezoelectric ceramic plates; 7. a diaphragm; 8. printing ink; 9. a nozzle; 10. ink particles; 321. a control end; 322. a sprocket arrangement; 323. a mechanical arm; 324. a robot arm.

Detailed Description

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.

In order to make the above objects, features and advantages of the present invention more comprehensible, the present invention is described in detail with reference to the accompanying drawings and the detailed description.

The utility model provides a 3D printing device, as shown in fig. 1-3, including computer 1, driving power supply 2, three-dimensional moving platform 3, printing shower nozzle 4, receiving platform 5. The computer 1 is wirelessly connected with the driving power supply 2, the computer 1 is internally provided with self-developed control software 11, and the control software 11 can control the disconnection of the switching printing channel voltage and set the working voltage and the printing drop number during printing. The connection port 21 of the driving power supply 2 is connected with the printing nozzle 3 through a wire, and can directly control the disconnection and connection of each printing channel of the printing nozzle 3 to provide proper working voltage for the printing nozzle 3. The printing nozzle 3 is internally provided with 5 independent printing channels, each corresponding printing channel is internally provided with an ink bag 41, each ink bag 41 can be filled with different printing ink 8 materials by a 5mL injector, then the gas in the ink bag 41 is discharged and is packaged by a sealing machine, and the operation steps can realize the independent and simultaneous printing of multiple materials. The three-dimensional moving platform 3 comprises a controller 31 and a three-axis mechanical arm 32, wherein in the three-axis mechanical arm 32, the movement in the directions of an X axis, a Y axis and a Z axis is controlled by a chain wheel structure 322; the X axis is the forward and backward axis of the robot arm 323. The Y-axis is the in and out axis of the robot 323. The Z axis is a rising and falling axis of the robot arm 323. The print head 4 is fixed to a robot arm 324 of the robot arm 323, and coordinates of a print pattern model are input in advance to the controller 31, whereby the movement locus of the print head is controlled, and the patterning process of the liquid droplets on the receiving stage 5 is realized. The choice of the receiving platform 5 needs to be determined according to practical situations, including but not limited to the following materials, such as glass/silicon wafer/metal panel, etc.

The utility model discloses when printing device used, the voltage of printing in-process and single inkjet drop number were controlled by control software 11 in the computer 1. The X-axis, Y-axis, and Z-axis coordinates of the printing model are set in the controller 31 of the three-dimensional moving platform 3, and thus, the patterned printing of the cells can be realized.

The utility model discloses a realize the patterning of cell based on piezoelectric type inkjet printing method deposit cell, this piezoelectric type inkjet printing's principle is: the printing ink 8 liquid drops in the ink bag 41 are sprayed out from the nozzle 9 by utilizing the telescopic deformation behavior of the piezoelectric ceramic piece 6 to form ink particles 10, and the outer side of the piezoelectric ceramic piece 6 is provided with an insulated diaphragm 7; the specific process is as follows: the negative pressure generated by the air pressure controller balances the sprayed ink at the position of the nozzle 9; the driving power supply 2 applies a pulse waveform, and in the rising and continuous processes of pulse voltage, the piezoelectric ceramic piece 6 generates weak deformation, the deformation causes the piezoelectric ceramic piece 6 to contact the glass capillary wall of the nearby ink bag 41 and form a sound wave, so that the solution at the nozzle 9 is extruded and sprayed out; when the voltage is reduced, the piezoelectric ceramic plate 6 is relaxed due to slow deformation, the glass capillary of the ink bag 41 expands, the ink at the nozzle 9 is sunken to cut off the extruded solution, and the extruded solution is gradually gathered to form a single ink particle 10 drop under the action of the surface tension of the ink, so that the jet printing meeting a certain rule is realized, and the piezoelectric ink-jet printing device is wide in application range and high in printing speed.

The three-axis robot arm 32 is of a rectangular coordinate type, and the arm is composed of three orthogonal pairs of movements. The X-axis and the Z-axis are connected into a whole through a control end 321, the other end of the control end 321 is fixed with the printing nozzle 4 through a manipulator 324, and the three axes are controlled to move linearly by a chain wheel transmission belt of a chain wheel structure 322 under the driving of a motor. The coordinate of the X/Z axis is accurately input into the controller 31 of the three-dimensional moving platform 3, so that the up-down and left-right movement of the printing nozzle can be controlled, the Y axis controls the front-back movement of the printing platform, and the control of the printing track and the patterning of ink are achieved through the cooperation of the three axes. The part has simple structure and high precision of movement position.

The parallel circuit of the driving power supply 2 mainly drives the two high-voltage switches by the control signal, and then cuts the high-voltage signal into square waves to realize the adjustment and conversion of the driving voltage. In addition, 5 switches in the parallel circuit are respectively connected with 5 leads at the tail part of the printing nozzle, and each lead is connected with one printing channel, so that independent or cooperative work of each printing channel can be realized.

The utility model discloses utilize piezoelectric type inkjet printing technique to print the liquid drop on receiving platform 5 to realize the accurate control of liquid drop and the device of accurate positioning. The method has the advantages of simple operation, high printing precision, good repeatability and small pollution probability to the sample in the printing process, and effectively solves the problems of complicated structure, single printing mode and the like of the printing device.

The utility model discloses based on above-mentioned scheme the device provides a method of printing and cultivateing the cell, including following step:

cleaning the printing channel; injecting the cell printing suspension into the ink bag, sealing an inlet of the ink bag, and spraying the cell printing suspension onto the glass substrate by using a voltage-driven ink-jet printing head; the cell printing suspension comprises DMEM complete medium and substanceSodium alginate aqueous solution with the weight percentage content of 0.2 percent to 1.2 percent and cells to be printed; the volume ratio of the DMEM complete culture medium to the sodium alginate aqueous solution with the mass percentage content of 0.2% -1.2% is 1: 1; the concentration of the cells to be printed in the cell printing suspension was 1X 10⁶～9×10⁶one/mL.

The printing voltage is 50-500V, the printing interval is 100-300 mu m, and the number of printing drops is 1-9 drops; covering a polydimethylsiloxane channel on the glass substrate sprayed with the cell printing suspension, and sequentially introducing 0.2-0.3M CaCl from the sample inlet hole by using a micro-injection pump₂Aqueous solution and DMEM complete culture medium, at 35-40 deg.C and 15% -25% CO₂Culturing under the conditions of (1).

The utility model firstly cleans the printing channel; before the printing operation is started, the utility model adds deionized water, ethanol and PBS buffer solution into the ink sac cavity in sequence to flush the printing channel and remove air bubbles, thereby preventing the blockage during the printing; the adding mode is preferably injected by a syringe; the PBS buffer is preferably 1 XPBS buffer; the pH value of the PBS buffer solution is preferably 7.4; the function of the PBS buffer was to rinse off the residual ethanol in the printing channels.

Cleaning the printing channel, the utility model injects the cell printing suspension into the ink bag 41, seals the inlet of the ink bag 41, and the voltage drives the ink-jet printing head to spray the cell printing suspension on the glass substrate; the cell printing suspension comprises a DMEM complete culture medium, a sodium alginate aqueous solution with the mass percentage content of 0.2% -1.2% and cells to be printed; the volume ratio of the DMEM complete culture medium to the sodium alginate aqueous solution with the mass percentage content of 0.2% -1.2% is 1: 1; the concentration of the cells to be printed in the cell printing suspension is 1 x 10⁶～9×10⁶Per mL; the mass percentage content of the sodium alginate aqueous solution is preferably 0.6-1%, and more preferably 0.8%; the printing voltage is 50-500V, preferably 100-400V, more preferably 200-300V, the printing interval is 100-300 μm, preferably 200 μm, the number of printing drops is 1-9 drops, preferably 2-6 drops, more preferably 3-5 drops.

The utility model discloses in, add in the medium and have sodium alginate aquogel, because there is the aperture of certain size in the gel granule, so allow the required nutrient substance of cell culture to pass through, provide abundant nutrient and good physiological environment for the cell.

The utility model discloses in, because the utility model discloses the separation principle of well cell is based on piezoelectric ink-jet printing, so printing voltage is different, the diameter variation in size of sodium alginate gel. In addition, the number of printed drops not only directly determines the diameter and volume of a single sodium alginate hydrogel, but also has a great influence on the environment of cell culture and the occupancy rate of cells. As the diameter of a single Hela cell is 10-20 μm, the diameter of a single gel particle is too large, and a plurality of cells can be contained, thus being not beneficial to single cell separation. Therefore, the size of the gel diameter under different voltages needs to be considered so as to provide a basis for setting the printing parameters of subsequent experiments.

In the utility model, the cells to be printed are preferably cultured by the following method: and (3) after the coverage rate of the cultured cells reaches over 80%, performing enzymolysis on the cells from the culture dish by using a trypsin aqueous solution, washing by using a culture medium, and performing centrifugal collection to obtain the cells to be printed.

In the present invention, the preparation of the glass substrate preferably includes: mixing glass and trichlorosilane, vacuumizing to-0.8 MPa, and keeping the vacuum state for 0.8-1.2 h, preferably 1h under the condition of-0.8 MPa to obtain the glass substrate.

Before mixing the glass and the trichlorosilane, the utility model preferably comprises the steps of washing the glass by deionized water and alcohol in sequence and drying; the drying mode is preferably blow-drying by nitrogen; the device for mixing the glass and the trichlorosilane is preferably a glass dryer; the vacuum state is kept for 0.8-1.2 h under the condition of-0.8 MPa, Si-OH groups on the surface are removed, the hydrogen bonding capability of the glass is reduced, the surface of the glass is hydrophobic, the three-dimensional shape of the sodium alginate gel is ensured when the sodium alginate gel is attached to the glass substrate, and the subsequent single cell research is facilitated. Meanwhile, the printed sodium alginate liquid drops are high in resolution ratio and not easy to displace when the vacuum state is kept for 0.8-1.2 h under the condition of-0.8 MPa, and formed gel liquid drops are regular.

The utility model discloses cover the polydimethylsiloxane passageway on the glass substrate that the spraying has cell to print suspension, utilize the micro-injection pump to let in 0.2 ~ 0.3M's CaCl from advancing the appearance hole in proper order₂Aqueous solution and DMEM complete culture medium, at 35-40 deg.C and 15% -25% CO₂Culturing under the conditions of (1); the polydimethylsiloxane channel preferably has a height of 20 to 100 μm, more preferably 35 to 50 μm, and a width of 0.6 to 1.2mm, preferably 0.8 to 1 mm.

The utility model discloses a print the adjustment of dropping number isoparametric, can control the number of the cell in the single gel of printing. The printed single cells still keep good cell activity and continuously grow in the culture device. The device and the cell separation method realize the long-term culture of the single cells. In the sodium alginate gel cell culture chamber, the cells are wrapped by the sodium alginate gel, and the three-dimensional environment that the cells are wrapped by extracellular matrix in vivo is also well simulated.

The utility model also provides an utilize micro-fluidic chip device and luminescent probe survey cell damage prosthetic method. The utility model uses the luminescent probe to detect the cell reproduction rate, the cell apoptosis and the change of the Reactive Oxygen Species (ROS) in the cell; the luminescent probe is a probe capable of respectively and specifically recognizing reactive oxygen species in cells of cell reproduction and cell apoptosis. Wherein the content change of the active oxygen can indirectly indicate cell damage and repair. Cell proliferation, apoptosis and changes in intracellular reactive oxygen species levels are several of the common indicators for detecting cellular activity and drug toxicity, and a decrease in intracellular reactive oxygen species indicates repair of cellular damage or a decrease in drug toxicity.

And (3) detecting cell indexes in a cell culture period, introducing an ethylene diamine tetraacetic acid (EDTA-2Na) solution into a channel to dissolve sodium alginate gel, collecting and centrifuging the solution to obtain cells, dyeing the cells, incubating for a period of time, detecting by using an enzyme-labeling instrument or imaging and photographing under a fluorescence microscope, and analyzing.

The utility model discloses an ink jet printing head is through voltage drive cell liquid drop injection on the glass substrate. After covering the PDMS layer, the sample inlet hole of the PDMS isSequentially and slowly injecting 0.2M CaCl by using a micro-injection pump₂Solution and cell high-sugar medium DMEM to achieve continuous long-term culture of cells.

The cell culture device comprises two parts, namely a glass (or silicon wafer) substrate and a PDMS layer. Firstly, a glass substrate with proper hydrophobicity is obtained by chemical treatment, and the main steps are as follows: the untreated glass sheets were cleaned with deionized water and ethanol, then blow dried with nitrogen, placed in a glass desiccator, 20 μ L of trichlorosilane was added, sealed and evacuated to about-0.8 MPa, and the vacuum was maintained for various periods of time. The effects of the above treatment times on gel spreading and adhesion and print resolution were compared. The printed sodium alginate liquid drops have high resolution ratio and are not easy to displace, and the formed gel liquid drops are regular.

Because the utility model discloses the separation principle of well cell is based on piezoelectricity inkjet printing, so printing voltage is different, the diameter variation in size of sodium alginate gel. In addition, the number of printed drops not only directly determines the diameter and volume of a single sodium alginate hydrogel, but also has a great influence on the environment of cell culture and the occupancy rate of cells. As the diameter of a single Hela cell is 10-20 μm, the diameter of a single gel particle is too large, and a plurality of cells can be contained, thus being not beneficial to single cell separation. Therefore, the size of the gel diameter under different voltages needs to be considered so as to provide a basis for setting the printing parameters of subsequent experiments. When the printing voltage is 50-500V and the number of drops is less than 10, the diameter of the printed single gel is relatively stable on the whole and is within 300 mu m.

Digesting the cells overgrowing in the cell culture bottle by pancreatin, adding fresh culture solution DMEM, blowing and beating the adherent cells by a liquid transfer gun, centrifuging the cell suspension for 3min under the condition of 1000r/min, removing the supernatant, and staining for 10min by using 2 mu M calcein-AM. Finally, the cells were resuspended in mixed culture medium to a concentration of 106 cells/mL. The mixed culture medium is formed by mixing a sodium alginate solution and a cell high-sugar culture medium DMEM in equal volume, and the final concentration of the sodium alginate is 0.4%. And after printing is finished each time, placing the glass sheet under a fluorescence microscope for observation and photographing, and counting the cell occupancy rate.

Hoechst 33342 is a blue fluorescent dye that can penetrate cell membranes, has low toxicity to cells, and is generally used for apoptosis detection. We observed apoptosis of the printed cells by staining the cells in advance with a fluorescence microscope. The results indicated that printed cells exhibited blue fluorescence, indicating that cells may be damaged during printing and have a tendency to undergo apoptosis. After a period of culture, the blue fluorescence value decreases, indicating that the cell damage is repaired to a certain extent.

And (3) detecting the cell proliferation rate by using a cck-8 apoptosis detection kit when culturing for 6h, 18h and 24h respectively, and measuring and analyzing the result by using an enzyme-labeling instrument. It can be seen that the cells had good proliferation activity within 6h of printing. After 18h, the cell proliferation rate is reduced, and at 24h, the cell proliferation rate is greatly improved, which shows that the activity of the cells is gradually recovered in the process, the adaptability of the cells to the environment is improved, the apoptosis number is reduced, and the cell proliferation rate is improved.

The fluorescence of the active oxygen of the cell which is just printed is stronger, which indicates that the level of the active oxygen in the cell is higher at the moment. The reason for this analysis may be that the printing process causes some damage to the cells, and apoptosis of the cells is caused by the high concentration of reactive oxygen species in the cells. This is also consistent with the results of Hoechst 33342 staining described above. After 12h of culture, the fluorescence of the active oxygen is greatly weakened, which shows that after the culture, the damage caused by cell printing is repaired, and the activity of the cells is recovered, so that the level of the active oxygen of the cells is greatly reduced; after 24h, the fluorescence intensity of the active oxygen detection is slightly reduced, which indicates that the active oxygen level in the cell begins to tend to be stable, namely the cell is adapted to the surrounding environment. Our analysis of cell damage and repair studies in this process indicates that the method can be used for the study of cell damage caused by membrane potential changes. The application of voltage causes a change in the potential of the outer mitochondrial membrane within the cell, which activates voltage-dependent anion channels on the outer mitochondrial membrane, forming hydrophilic channels on the membrane. Breaks the intracellular metabolic balance and calcium homeostasis, generates precursor superoxide anion (O2-) synthesized by ROS, sharply increases the intracellular ROS content, promotes the Ca2+ influx, thereby up-regulating the expression of bax, MPTP opening and Caspase activation, and leading to apoptosis. Thereby causing the cell to have a tendency to undergo apoptosis. In the subsequent culture process, the cells start a self-repair mechanism, the intracellular steady state is gradually recovered under a good growth environment, the intracellular ROS content is reduced, and the cells gradually recover the activity.

The utility model discloses a concrete example is applied to explain the principle and the implementation mode of the utility model, and the explanation of the above example is only used to help understand the method and the core idea of the utility model; meanwhile, for the general technical personnel in the field, according to the idea of the present invention, there are changes in the concrete implementation and the application scope. In summary, the content of the present specification should not be construed as a limitation of the present invention.

Claims

1. The utility model provides a 3D printing device which characterized in that: the three-dimensional mobile platform is provided with a three-axis mechanical arm, the three-axis mechanical arm is electrically connected with a controller, a printing nozzle is arranged on the three-axis mechanical arm, the printing nozzle is electrically connected with a driving power supply, the driving power supply is wirelessly connected with a computer, and control software is arranged in the computer; and a receiving platform is fixedly arranged on the three-dimensional moving platform, the receiving platform is positioned below the printing nozzle, and printing ink is filled in the printing nozzle.

2. The 3D printing device according to claim 1, characterized in that: a plurality of printing channels are uniformly arranged in the printing nozzle, an ink bag is arranged in each printing channel, and the printing ink is contained in the ink bag; the bottom of the ink bag is provided with a nozzle, the side wall of the ink bag is provided with a piezoelectric ceramic piece, and the printing channel is electrically connected with the driving power supply through a wire; and a diaphragm is arranged on the outer side of the piezoelectric ceramic piece.

3. The 3D printing device according to claim 1, characterized in that: triaxial arm include the level fixed set up in Y axle on the three-dimensional moving platform, the epaxial Z axle that slides and is provided with vertical installation of Y, Z axle top fixed mounting has the X axle that the level set up, just the horizontal projection of X axle with the horizontal projection of Y axle sets up perpendicularly, be provided with the control end through sprocket structure horizontal slip on the X axle, be provided with the arm through the vertical slip of sprocket structure on the control end lateral wall, the terminal fixed mounting of arm has the manipulator, print the shower nozzle install in on the manipulator.

4. The 3D printing device according to claim 2, characterized in that: the driving power supply is provided with a connecting port, and the connecting port is connected with the printing channel through a lead.

5. The 3D printing device according to claim 2, characterized in that: five printing channels are uniformly arranged in the printing nozzle, each printing channel is internally provided with an ink bag, and five ink bags are respectively filled with different printing inks by using 5mL injectors.

6. The 3D printing device according to claim 4, characterized in that: the driving power supply adopts a parallel circuit, the parallel circuit adopts a control signal to drive two high-voltage switches, and the high-voltage signals are cut into square waves to realize the adjustment and conversion of the magnitude of the driving voltage; the parallel circuit is provided with a plurality of switches in parallel, and each switch is connected with one printing channel through a lead.