Device and method for generating dynamic chemical signal by using piezoelectric diaphragm pump

Abstract

The invention discloses a device and a method for generating a dynamic chemical signal by using a piezoelectric diaphragm pump, and aims to improve the precision and the efficiency of generating a drug dynamic signal in a cell biology research. The device is composed of a dynamic chemical signal generation assembly and a microfluid mixing chip, wherein the dynamic chemical signal generation assembly comprises a piezoelectric diaphragm pump, a piezoelectric diaphragm pump control module and a piezoelectric diaphragm pump control module frame; the piezoelectric diaphragm pump can realize accurate flow control, and supports parallel connection or series connection of multiple pumps to increase the flow; the microfluid mixing chip is provided with a plurality of liquid inlets, and uniform mixing is realized through a plurality of rows of mixing channels; liquid pumping can be controlled in a full-automatic mode, and manual operation is reduced. The cell experiment table is small in overall design, portable, suitable for being used on a table top and capable of providing convenience for cell experiments. According to the invention, a single or complex dynamic chemical signal can be effectively generated, a cell microenvironment can be accurately simulated, and the method has important significance on cell behavior research.

Description

Technical Field

[0001] The invention belongs to the technical field of cell biology, and in particular relates to a device and method for generating dynamic chemical signals by using a piezoelectric diaphragm pump. Background Art

[0002] In the study of cell biology, adding drugs to cells to study cell behavior is a common experimental method. This method can help scientists explore the effects of drugs on cell physiological and pathological processes, including basic life activities such as cell proliferation, differentiation, apoptosis, and movement. Through these studies, scientists can reveal the signaling network inside cells and its mechanism of action, understand how cells respond to external signals, and how these responses affect the fate and function of cells.

[0003] In traditional methods, this process usually requires manual operation, which is inefficient and easily affected by the operator's skill level. At present, there are devices such as syringe pumps for precise drug addition, but the existing injection devices generally have the problem of large size. Ordinary drug addition titration experiments only add a certain amount of drugs at regular intervals to achieve drug dynamic signals, and cannot accurately control the concentration of the cell environment, and the generated signal is not a standard dynamic signal. Moreover, whether it is manual titration drug addition experiments or drug addition experiments using syringe pump equipment, it is often impossible to accurately add trace drugs. However, cells are extremely sensitive to the living environment, and the large difference in drug content added each time may affect cell behavior. Summary of the invention

[0004] In view of the above-mentioned problems, the object of the present invention is to provide a device and method for generating dynamic chemical signals using a piezoelectric diaphragm pump.

[0005] The technical solution of the present invention is: a device for generating dynamic chemical signals using a piezoelectric diaphragm pump, comprising a dynamic chemical signal generating component and a microfluidic mixing chip;

[0006] The dynamic chemical signal generating component comprises a piezoelectric diaphragm pump control module frame and a plurality of piezoelectric diaphragm pump control modules arranged on the piezoelectric diaphragm pump control module frame, each of the piezoelectric diaphragm pump control modules having a piezoelectric diaphragm pump therein;

[0007] The piezoelectric diaphragm pump control module includes a box-shaped shell, a piezoelectric diaphragm pump slot and a piezoelectric diaphragm pump control board arranged in the box-shaped shell, the piezoelectric diaphragm pump is fixedly connected in the piezoelectric diaphragm pump slot and is control-connected to the piezoelectric diaphragm pump control board, and two pipeline connection holes are arranged on the side of the box-shaped shell;

[0008] The piezoelectric diaphragm pump has an input end, an output end and a control interface. The input end is connected to an input pipeline, and the input pipeline passes through one of the pipeline connecting holes to communicate with the external liquid. The output end is connected to an output pipeline, and the output pipeline passes through another of the pipeline connecting holes to connect to the microfluidic mixing chip. The control interface is control-connected to the piezoelectric diaphragm pump control board.

[0009] Furthermore, the microfluidic mixing chip comprises:

[0010] Carrier board;

[0011] A plurality of chip liquid inlets are arranged at one end of the carrier plate, and the output pipelines are respectively connected to the chip liquid inlets correspondingly;

[0012] A merging cavity, disposed on the carrier plate, connected to the output ports of the plurality of chip liquid inlets;

[0013] A liquid mixing channel, wherein multiple rows of plates are arranged on the carrier plate, one end of the liquid mixing channel is connected to the confluence cavity, and the other end is provided with a drainage port, and the drainage port is connected to an external cell culture device through a drainage pipeline;

[0014] The observation chamber is arranged on the liquid mixing channel near the liquid discharge port.

[0015] Furthermore, the piezoelectric diaphragm pump control module frame includes:

[0016] A partition fixing frame, on which a plurality of the piezoelectric diaphragm pump control modules are arranged at intervals, and the number of the piezoelectric diaphragm pump control modules is 1-4;

[0017] Rubber shock absorber, a plurality of rubber shock absorbers are arranged below the partition fixing frame.

[0018] Furthermore, the box-shaped housing comprises:

[0019] A control board fixing hole is arranged at the bottom of the box-shaped housing, and the piezoelectric diaphragm pump control board is fixed on the control board fixing hole;

[0020] The cover plate fixing hole is arranged at the edge of the open end of the box-shaped shell;

[0021] A transparent cover plate, which is disposed on the box-shaped shell and is fixedly connected to the box-shaped shell through the fixing holes of the cover plate;

[0022] The power supply hole and the communication interface are arranged on the side of the box-shaped shell, and the power supply hole and the communication interface are respectively connected to the piezoelectric diaphragm pump control board.

[0023] Further,

[0024] The box-shaped shell is made of any one of photosensitive resin, aluminum alloy or ABS plastic material;

[0025] The transparent cover is made of ABS plastic material;

[0026] The piezoelectric diaphragm pump control module frame is made of any one of photosensitive resin, aluminum alloy or ABS plastic material;

[0027] The microfluid mixing chip is made of either PDMS polydimethylsiloxane or ABS plastic material.

[0028] The working method of the device for generating dynamic chemical signals using a piezoelectric diaphragm pump comprises the following steps:

[0029] The input end of the piezoelectric diaphragm pump is connected to the external liquid through the input pipeline, the output end is connected to the chip liquid inlet of the microfluidic mixing chip through the output pipeline, the power supply is connected through the power supply hole, and the communication interface is connected to the external computer control device;

[0030] The number of piezoelectric diaphragm pumps to be operated is selected and numbered through an external computer control device, and corresponding instructions are sent to the corresponding piezoelectric diaphragm pumps, so that the input line pipe and the output line pipe connected to the corresponding piezoelectric diaphragm pump are filled with the above-mentioned liquid;

[0031] The operation mode of each piezoelectric diaphragm pump is set by an external computer control device: and corresponding parameters are input to the piezoelectric diaphragm pump according to the selected mode;

[0032] Each piezoelectric diaphragm pump is operated to pass the liquid into the merging cavity through the liquid inlet of the chip respectively. After the liquid is merged in the merging cavity, it enters the liquid mixing channel for mixing. The evenly mixed liquid flows into the external cell culture device through the liquid outlet of the chip;

[0033] By setting the operating mode of the piezoelectric diaphragm pump, any fluid output can be achieved, including but not limited to:

[0034] Liquid sinusoidal output mode: The parameters input in this mode include the initial concentration of the drug controlled by the piezoelectric diaphragm pump, the peak value of the required target concentration waveform, the cycle time, the initial phase and the total running time of the piezoelectric diaphragm pump, which can achieve sinusoidal output of the drug concentration in the liquid;

[0035] Liquid constant concentration output mode: The parameters input in this mode include the initial concentration of the drug controlled by the piezoelectric diaphragm pump, the required target concentration and the total running time of the piezoelectric diaphragm pump, which can achieve constant concentration output of the drug in the liquid;

[0036] Liquid pulse output mode: The parameters input in this mode include the initial concentration of the drug controlled by the piezoelectric diaphragm pump, the required target concentration, cycle time, duration within the waveform cycle and the total running time of the piezoelectric diaphragm pump, which can achieve constant concentration pulse output of drugs in liquids;

[0037] After inputting corresponding parameters to each piezoelectric diaphragm pump according to the selected liquid output mode, the total flow of all piezoelectric diaphragm pumps is input into the external computer control device to run the calculation to see whether the input parameters reach the target parameters;

[0038] If not, a pop-up window will inform you that the input parameters cannot meet all target parameters at the same time, and ask whether to calculate the initial drug concentrations required to achieve the target parameters. If you select yes, the initial drug concentrations required to achieve the target parameters will be calculated. After the calculation is completed, a pop-up window will prompt the required initial concentrations of each drug. If you select no, exit the pop-up window;

[0039] If so, an image of the flow rate variation over time of the selected piezoelectric diaphragm pump and an image of the target concentration of each drug variation over time are popped up.

[0040] Compared with the prior art, the present invention has the following beneficial effects:

[0041] (1) In the device for generating dynamic chemical signals using a piezoelectric diaphragm pump of the present invention, the flow is driven by a piezoelectric diaphragm pump instead of a syringe or an injection pump; taking water as an example, when driven by a voltage of 250V, the flow rate of a single pump ranges from 200μL to 5000μL; the device can also use a series or parallel piezoelectric diaphragm pump to increase the flow rate, which facilitates the regulation of the cell microenvironment.

[0042] (2) In the device for generating dynamic chemical signals using a piezoelectric diaphragm pump of the present invention, the flow rate is driven by a piezoelectric diaphragm pump to pump the liquid. Under the condition that the voltage of the piezoelectric diaphragm pump and the duty cycle of the PWM wave of the signal output by the piezoelectric diaphragm pump control board remain unchanged, the flow rate pumped by the piezoelectric diaphragm pump each time is the same and controllable. The flow rate can be changed at any time by changing the voltage of the piezoelectric diaphragm pump, the duty cycle of the PWM wave signal output by the piezoelectric diaphragm pump control board, and the frequency of the PWM wave signal output by the piezoelectric diaphragm pump control board to achieve dynamic changes in the output of drugs in the liquid. The three typical modes are: sinusoidal output, constant concentration output, and pulse output mode. The dynamic output of drugs (chemical signals) of any concentration in the liquid through a single mode or a combination of multiple modes provides convenient conditions for the generation of dynamic chemical signals in biological research.

[0043] (3) In the device of the present invention for generating dynamic chemical signals using a piezoelectric diaphragm pump, the piezoelectric diaphragm pump can be automatically controlled to pump liquid, thereby reducing manual operations and increasing convenience during use; and the device can be added with no more than seven types of drugs, thereby increasing the diversity and complexity of the experiment and providing the possibility of simulating the cellular microenvironment in the human body.

[0044] (4) In the device for generating dynamic chemical signals using a piezoelectric diaphragm pump of the present invention, the overall device is relatively small compared to other injection devices on the market. A single piezoelectric diaphragm pump control module is 110 mm long, 90 mm wide, and 30 mm high, and the overall device is 130 mm long, 110 mm wide, and 110 mm high. The overall device is simple and portable and can be used as a desktop device, providing convenience for cell experiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 It is a schematic diagram of the overall structure of a device for generating dynamic chemical signals using a piezoelectric diaphragm pump according to the present invention;

[0046] Figure 2 It is a schematic diagram of the structure of the piezoelectric diaphragm pump control module in the present invention;

[0047] Figure 3 It is a schematic diagram of the structure of the piezoelectric diaphragm pump control module frame in the present invention;

[0048] Figure 4 is a schematic diagram of a piezoelectric diaphragm pump in the present invention;

[0049] Figure 5 is a schematic diagram of the microfluidic mixing chip of the present invention;

[0050] Figure 6 is the change in fluorescence intensity within one cycle in Case 1 of the present invention;

[0051] Figure 7 is the normalized fluorescence intensity change during the total duration in Case 1 of the present invention;

[0052] Figure 8 These are the changes in the two fluorescence intensities in the total time in Case 2 of the present invention (isothiocyanate on the left and rhodamine on the right).

[0053] Among them, 1. dynamic chemical signal generating component; 2. microfluidic mixing chip; 3. piezoelectric diaphragm pump; 4. piezoelectric diaphragm pump control module; 5. piezoelectric diaphragm pump control module rack; 6. box-shaped shell; 7. transparent cover; 8. piezoelectric diaphragm pump control board; 9. piezoelectric diaphragm pump slot; 10. control board fixing hole; 11. cover fixing hole; 12. pipeline connection hole; 13. power supply hole; 14. communication interface; 15. input end; 16. output end; 17. control interface; 18. rubber shock absorber; 19. partition fixing frame; 20. chip liquid inlet; 21. confluence cavity; 22. liquid mixing channel; 23. observation cavity; 24. chip liquid outlet. DETAILED DESCRIPTION

[0054] The following is combined with Figure 1 To the attached Figure 8 , the specific embodiments of the present invention are described in detail. In the description of the present invention, it should be understood that the terms "center", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention.

[0055] The terms "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, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features; in the description of the present invention, unless otherwise specified, "plurality" means two or more.

[0056] It should be noted that the circuit connections involved in the present invention all adopt conventional circuit connection methods and do not involve any innovation.

[0057] Example: Figure 1 As shown, a device for generating dynamic chemical signals using a piezoelectric diaphragm pump comprises a dynamic chemical signal generating component 1 and a microfluidic mixing chip 2;

[0058] The dynamic chemical signal generating component 1 includes a piezoelectric diaphragm pump control module frame 5 and a plurality of piezoelectric diaphragm pump control modules 4 arranged on the piezoelectric diaphragm pump control module frame 5, each piezoelectric diaphragm pump control module 4 is provided with a piezoelectric diaphragm pump 3;

[0059] like Figure 2As shown, the piezoelectric diaphragm pump control module 4 includes a box-shaped housing 6, a piezoelectric diaphragm pump slot 9 and a piezoelectric diaphragm pump control board 8 arranged in the box-shaped housing 6, the piezoelectric diaphragm pump 3 is fixedly connected in the piezoelectric diaphragm pump slot 9, and is controlled and connected with the piezoelectric diaphragm pump control board 8, and two pipeline connection holes 12 are arranged on the side of the box-shaped housing 6;

[0060] like Figure 4 As shown, the piezoelectric diaphragm pump 3 has an input end 15, an output end 16 and a control interface 17, the input end 15 is connected with an input pipeline, the input pipeline passes through a pipeline connection hole 12 and is in communication with an external liquid, the output end 16 is connected with an output pipeline, the output pipeline passes through another pipeline connection hole 12 and is connected to the microfluidic mixing chip 2, and the control interface 17 is control-connected to the piezoelectric diaphragm pump control board 8;

[0061] like Figure 5 As shown, the microfluidic mixing chip 2 includes a carrier, a chip liquid inlet 20, a confluence cavity 21, a liquid mixing channel 22 and an observation cavity 23; a plurality of chip liquid inlets 20 are arranged at one end of the carrier, and the output pipelines are respectively connected to the chip liquid inlets 20; the confluence cavity 21 is arranged on the carrier, and the confluence cavity 21 is connected to the output ports of the plurality of chip liquid inlets 20; a plurality of rows of liquid mixing channels 22 are arranged on the carrier, one end of the liquid mixing channel 22 is connected to the confluence cavity 21, and the other end is provided with a drain port 24, and the drain port 24 is connected to an external cell culture device through a drain pipeline; the observation cavity 23 is arranged on the liquid mixing channel 22 near the drain port 24.

[0062] Preferably, if Figure 3 As shown, the piezoelectric diaphragm pump control module frame 5 includes a partition fixing frame 19 and a rubber shock absorber 18; multiple piezoelectric diaphragm pump control modules 4 are arranged at intervals on the partition fixing frame 19, and the number of piezoelectric diaphragm pump control modules 4 is 4; multiple rubber shock absorbers 18 are arranged under the partition fixing frame 19.

[0063] Preferably, the box-shaped shell 6 includes a control board fixing hole 10, a cover plate fixing hole 11, a transparent cover plate 7, a power supply hole 13 and a communication interface 14; the control board fixing hole 10 is arranged at the bottom of the box-shaped shell 6, and the piezoelectric diaphragm pump control board 8 is fixed on the control board fixing hole 10; the cover plate fixing hole 11 is arranged at the edge of the open end of the box-shaped shell 6; the transparent cover plate 7 is covered on the box-shaped shell 6, and is fixedly connected to the box-shaped shell 6 through the cover plate fixing hole 11; the power supply hole 13 and the communication interface 14 are arranged on the side of the box-shaped shell 6, and the power supply hole 13 and the communication interface 14 are respectively connected to the piezoelectric diaphragm pump control board 8.

[0064] Preferably, the box-shaped housing 6 is made of photosensitive resin; the transparent cover plate 7 is made of ABS plastic material; the piezoelectric diaphragm pump control module frame 5 is made of aluminum alloy; and the microfluidic mixing chip 2 is made of PDMS polydimethylsiloxane.

[0065] The working method of the above embodiment comprises the following steps:

[0066] The input end 15 of the piezoelectric diaphragm pump 3 is connected to the external liquid through the input pipeline, the output end 16 is connected to the chip liquid inlet 20 of the microfluidic mixing chip 2 through the output pipeline, the power supply is connected through the power supply hole 13, and the communication interface 14 is connected to the external computer control device;

[0067] The number of piezoelectric diaphragm pumps 3 to be operated is selected and numbered through an external computer control device, and corresponding instructions are sent to the corresponding piezoelectric diaphragm pumps 3, so that the input line pipe and the output line pipe connected to the corresponding piezoelectric diaphragm pump 3 are filled with the above-mentioned liquid;

[0068] The operation mode of each piezoelectric diaphragm pump 3 is set by an external computer control device: and corresponding parameters are input to the piezoelectric diaphragm pump 3 according to the selected mode;

[0069] Each piezoelectric diaphragm pump 3 is in operation, and the liquid is respectively introduced into the merging cavity 21 through the liquid inlet 20 of the chip. After the liquid is merged in the merging cavity 21, it enters the liquid mixing channel 22 for mixing. The evenly mixed liquid flows into the external cell culture device through the liquid outlet 24 of the chip.

[0070] The operating mode of the piezoelectric diaphragm pump 3 is set to include:

[0071] Liquid sinusoidal output mode: The parameters input in this mode include the initial concentration of the drug controlled by the piezoelectric diaphragm pump 3, the peak value of the required target concentration waveform, the cycle time, the initial phase and the total running time of the piezoelectric diaphragm pump 3;

[0072] Liquid constant concentration output mode: The parameters input in this mode include the initial concentration of the drug controlled by the piezoelectric diaphragm pump 3, the required target concentration and the total running time of the piezoelectric diaphragm pump 3;

[0073] Liquid pulse output mode: The parameters input in this mode include the initial concentration of the drug controlled by the piezoelectric diaphragm pump 3, the required target concentration, the cycle time, the duration within the waveform cycle and the total operating time of the piezoelectric diaphragm pump 3;

[0074] After inputting corresponding parameters to each piezoelectric diaphragm pump 3 according to the selected liquid output mode, the total flow of all piezoelectric diaphragm pumps 3 is inputted into the external computer control device to run and calculate whether the input parameters reach the target parameters;

[0075] If not, a pop-up window will inform you that the input parameters cannot meet all target parameters at the same time, and ask whether to calculate the initial drug concentrations required to achieve the target parameters. If you select yes, the initial drug concentrations required to achieve the target parameters will be calculated. After the calculation is completed, a pop-up window will prompt the required initial concentrations of each drug. If you select no, exit the pop-up window;

[0076] If so, an image of the flow rate variation over time of the selected piezoelectric diaphragm pump 3 and an image of the target concentration of each drug variation over time are popped up.

[0077] Case 1: Single dynamic chemical signal generation

[0078] Take out two piezoelectric diaphragm pump control modules 4 , put one into the buffer control module position of the piezoelectric diaphragm pump control module rack 5 , and put the other into other random position of the piezoelectric diaphragm pump control module rack 5 . Connect the power supply holes 13 and the communication interface 14 of the two piezoelectric diaphragm pump control modules 4 to an external computer, wherein the input end 15 of the piezoelectric diaphragm pump 3 in one of the piezoelectric diaphragm pump control modules 4 is connected to a 0.01 mg / ml rhodamine solution through an input pipeline, and a command is sent to fill the piezoelectric diaphragm pump 3 and the input pipeline with the rhodamine solution, and the output end 16 of the piezoelectric diaphragm pump 3 is connected to any one of the chip liquid inlets 20 of the microfluidic mixing chip 2 through the output pipeline; the input end 15 of the piezoelectric diaphragm pump 3 in the other piezoelectric diaphragm pump control module 4 is connected to a buffer solution through an input pipeline, and the buffer solution uses deionized water, and a command is sent to fill the piezoelectric diaphragm pump 3 and the input pipeline with deionized water, and the output end 16 of the piezoelectric diaphragm pump 3 is connected to any other chip liquid inlet 20 of the microfluidic mixing chip through the output pipeline, and the other chip liquid inlets 20 are blocked with pi n needles to prevent leakage;

[0079] Place the microfluidic mixing chip 2 under an inverted fluorescence microscope, set the fluorescence channel to TRI TC, set the shooting time to five minutes, and the shooting interval to 500ms, and observe the instantaneous fluorescence intensity at the observation chamber 23 of the microfluidic mixing chip 2;

[0080] Open the fluid mixing software, select the number of the piezoelectric diaphragm pump control module 4 for inputting the rhodamine solution, then select the mode of the piezoelectric diaphragm pump control module 4 for inputting the rhodamine solution as the sinusoidal input mode, input the initial drug concentration as 0.01 mg / ml, input the target sinusoidal generated concentration amplitude as 0.002 mg / ml, the initial phase as zero, the period as 32 s, and the total time as 5 minutes; select the number of the piezoelectric diaphragm pump control module 4 for the buffer solution, set the instantaneous maximum total flow rate to 10 ml / min, and click Start to generate a single chemical signal;

[0081] The instantaneous fluorescence intensity was observed under an inverted fluorescence microscope. Figure 6 , 7As shown, the resulting intensity is a sinusoidal waveform over the total time.

[0082] Case 2: Generation of multiple complex dynamic chemical signals

[0083] Take out three piezoelectric diaphragm pump control modules 4, put one into the buffer control module position of the piezoelectric diaphragm pump control module rack 5, and put the other two into random positions of the piezoelectric diaphragm pump control module rack 5; connect the power supply holes 13 and the communication interface 14 of the three piezoelectric diaphragm pump control modules 4 to the external computer, connect the input end 15 of the piezoelectric diaphragm pump 3 in the first piezoelectric diaphragm pump control module 4 to the 0.01mg / ml rhodamine solution through an input pipeline, and connect the input end 15 of the piezoelectric diaphragm pump 3 in the second piezoelectric diaphragm pump control module 4 to the 0.005mg / ml fluorescein isothiocyanate solution through an input pipeline, and send instructions to make the piezoelectric diaphragm pump 3 and the input The inside of the pipeline is filled with rhodamine solution and fluorescein isothiocyanate solution respectively, and then the output ends 16 of the two piezoelectric diaphragm pumps 3 are connected to any two chip liquid inlets 20 of the microfluidic mixing chip 2 through the output pipeline; the input end 15 of the piezoelectric diaphragm pump 3 in another piezoelectric diaphragm pump control module 4 is connected to the buffer solution through the input pipeline, and the buffer solution uses deionized water. An instruction is sent to fill the piezoelectric diaphragm pump 3 and the input pipeline with deionized water, and the output end 16 of the piezoelectric diaphragm pump 3 in this piezoelectric diaphragm pump control module 4 is connected to any other chip liquid inlet 20 of the microfluidic mixing chip 2 through the output pipeline, and the other chip liquid inlets 20 are blocked with pin needles to prevent leakage;

[0084] The microfluidic mixing chip 2 is placed under an inverted fluorescence microscope, and the fluorescence channels are set to TRI TC and FI TC dual channels for simultaneous shooting. The shooting time is set to 60 minutes, and the shooting interval is set to 5 seconds. The instantaneous fluorescence intensity is observed at the observation chamber 23 of the microfluidic mixing chip 2;

[0085] Open the fluid mixing software, switch to the complex fluid generation mode, select the number of the piezoelectric diaphragm pump control module 4 for inputting the rhodamine solution, and then select the mode of the piezoelectric diaphragm pump control module 4 for inputting the rhodamine solution as the sinusoidal input mode, input the initial drug concentration as 0.01 mg / ml, input the sinusoidal maximum value as 4500 μL / Min, the minimum value as 1000 μL / Min, the initial phase as zero, the period as 300s, and the total time as 60 minutes; select the number of the piezoelectric diaphragm pump control module 4 for inputting the fluorescein isothiocyanate solution, and then select the mode of the piezoelectric diaphragm pump control module 4 for inputting the rhodamine solution as the sinusoidal input mode, input the initial drug concentration as 0.005 mg / ml, input the sinusoidal maximum value as 3500 μL / Min, the minimum value as 500 μL / Min, the initial phase as zero, the period as 180s, and the total time as 60 minutes; select the number of the piezoelectric diaphragm pump control module 4 for the buffer solution, set the mode to the constant frequency mode, and set the flow rate to 5000 μL / Mi n, click Start to start generating complex fluid chemical signals;

[0086] The instantaneous fluorescence intensity was observed under an inverted fluorescence microscope. Figure 8 As shown, the changes of the two fluorescence intensities over time are shown respectively.

[0087] The specific models of the above electronic components are not specially specified, and ordinary products available on the market can be selected as long as they can meet the use requirements of the present invention.

[0088] The specific embodiments described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above description is only a specific embodiment of the present invention and does not limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention are included in the protection scope of the present invention.

Claims

1. A device for generating dynamic chemical signals using a piezoelectric diaphragm pump, characterized in that: It comprises a dynamic chemical signal generating component (1) and a microfluidic mixing chip (2); The dynamic chemical signal generating component (1) comprises a piezoelectric diaphragm pump control module frame (5) and a plurality of piezoelectric diaphragm pump control modules (4) arranged on the piezoelectric diaphragm pump control module frame (5), each of the piezoelectric diaphragm pump control modules (4) being provided with a piezoelectric diaphragm pump (3); The piezoelectric diaphragm pump control module (4) comprises a box-shaped housing (6), a piezoelectric diaphragm pump slot (9) and a piezoelectric diaphragm pump control board (8) arranged in the box-shaped housing (6); the piezoelectric diaphragm pump (3) is fixedly connected in the piezoelectric diaphragm pump slot (9) and is control-connected to the piezoelectric diaphragm pump control board (8); and two pipeline connection holes (12) are arranged on the side of the box-shaped housing (6); The piezoelectric diaphragm pump (3) has an input end (15), an output end (16) and a control interface (17); the input end (15) is connected to an input pipeline, the input pipeline passes through one of the pipeline connection holes (12) to communicate with the external liquid; the output end (16) is connected to an output pipeline, the output pipeline passes through another of the pipeline connection holes (12) to connect to the microfluidic mixing chip (2); the control interface (17) is control-connected to the piezoelectric diaphragm pump control board (8).

2. A device for generating dynamic chemical signals using a piezoelectric diaphragm pump as claimed in claim 1, characterized in that: The microfluidic mixing chip (2) comprises: Carrier board; A plurality of chip liquid inlets (20) are arranged at one end of the carrier plate, and the output pipelines are respectively connected to the chip liquid inlets (20) correspondingly; A merging cavity (21) is arranged on the carrier plate, and the merging cavity (21) is connected to the output ports of the plurality of chip liquid inlets (20); A liquid mixing channel (22), wherein multiple rows of plates are arranged on the carrier plate, one end of the liquid mixing channel (22) is connected to the merging chamber (21), and the other end is provided with a drainage port (24), and the drainage port (24) is connected to an external cell culture device through a drainage pipeline; An observation chamber (23) is arranged on the liquid mixing channel (22) near the liquid discharge port (24).

3. A device for generating dynamic chemical signals using a piezoelectric diaphragm pump as claimed in claim 1, characterized in that: The piezoelectric diaphragm pump control module frame (5) comprises: A partitioning fixed frame (19), a plurality of the piezoelectric diaphragm pump control modules (4) are arranged at intervals on the partitioning fixed frame (19), and the number of the piezoelectric diaphragm pump control modules (4) is 1-4; A rubber shock absorber (18), wherein a plurality of the rubber shock absorbers (18) are arranged below the partition fixing frame (19).

4. A device for generating dynamic chemical signals using a piezoelectric diaphragm pump as claimed in claim 1, characterized in that: The box-shaped housing (6) comprises: A control board fixing hole (10) is arranged at the bottom of the box-shaped housing (6), and the piezoelectric diaphragm pump control board (8) is fixed on the control board fixing hole (10); A cover plate fixing hole (11) is arranged at the edge of the open end of the box-shaped housing (6); A transparent cover plate (7) is disposed on the box-shaped outer shell (6) and is fixedly connected to the box-shaped outer shell (6) via the cover plate fixing holes (11); A power supply hole (13) and a communication interface (14) are arranged on the side of the box-shaped housing (6), and the power supply hole (13) and the communication interface (14) are respectively connected to the piezoelectric diaphragm pump control board (8).

5. A device for generating dynamic chemical signals using a piezoelectric diaphragm pump as claimed in claim 1, characterized in that: The box-shaped housing (6) is made of any one of photosensitive resin, aluminum alloy or ABS plastic material; The transparent cover plate (7) is made of ABS plastic material; The piezoelectric diaphragm pump control module frame (5) is made of any one of photosensitive resin, aluminum alloy or ABS plastic material; The microfluid mixing chip (2) is made of either PDMS polydimethylsiloxane or ABS plastic material.

6. A method for operating a device for generating a dynamic chemical signal using a piezoelectric diaphragm pump according to claim 4, characterized in that: The steps include: The input end (15) of the piezoelectric diaphragm pump (3) is connected to an external liquid through an input pipeline, the output end (16) is connected to a chip liquid inlet (20) of a microfluidic mixing chip (2) through an output pipeline, a power supply is connected through a power supply hole (13), and a communication interface (14) is connected to an external computer control device; The number of piezoelectric diaphragm pumps (3) to be operated is selected and numbered through an external computer control device, and corresponding instructions are sent to the corresponding piezoelectric diaphragm pumps (3) respectively, so that the input line pipe and the output line pipe connected to the corresponding piezoelectric diaphragm pump (3) are filled with the above-mentioned liquid; The operating mode of each piezoelectric diaphragm pump (3) is set by an external computer control device, and corresponding parameters are input into the piezoelectric diaphragm pump (3) according to the selected mode; Each piezoelectric diaphragm pump (3) is in operation to pass liquid into the merging chamber (21) through the chip liquid inlet (20), and after the liquid is merged in the merging chamber (21), it enters the liquid mixing channel (22) for mixing, and the evenly mixed liquid flows into the external cell culture device through the chip liquid outlet (24).

7. A method for operating a device for generating a dynamic chemical signal using a piezoelectric diaphragm pump as claimed in claim 6, characterized in that: The operating mode of the piezoelectric diaphragm pump (3) includes: Liquid sinusoidal output mode: the parameters input in this mode include the initial concentration of the drug controlled by the piezoelectric diaphragm pump (3), the peak value of the desired target concentration waveform, the cycle time, the initial phase and the total operating time of the piezoelectric diaphragm pump (3); Liquid constant concentration output mode: the parameters input in this mode include the initial concentration of the drug controlled by the piezoelectric diaphragm pump (3), the required target concentration and the total running time of the piezoelectric diaphragm pump (3); Liquid pulse output mode: The parameters input in this mode include the initial concentration of the drug controlled by the piezoelectric diaphragm pump (3), the required target concentration, the cycle time, the duration within the waveform cycle and the total operating time of the piezoelectric diaphragm pump (3).

8. The working method of the device for generating dynamic chemical signals using a piezoelectric diaphragm pump as claimed in claim 6, characterized in that: After inputting corresponding parameters to each piezoelectric diaphragm pump (3) according to the selected mode, the total flow rate of all piezoelectric diaphragm pumps (3) is inputted into the external computer control device, and the operation is performed to calculate whether the input parameters reach the target parameters; If not, a pop-up window will inform you that the input parameters cannot meet all target parameters at the same time, and ask whether to calculate the initial drug concentrations required to achieve the target parameters. If you select yes, the initial drug concentrations required to achieve the target parameters will be calculated. After the calculation is completed, a pop-up window will prompt the required initial concentrations of each drug. If you select no, exit the pop-up window; If so, an image of the flow rate of the selected piezoelectric diaphragm pump (3) changing over time and an image of the target concentration of each drug changing over time are popped up.

Images