Device and method for generating dynamic chemical signals using a piezoelectric diaphragm pump

Abstract

The present invention discloses a device and method for generating dynamic chemical signals using a piezoelectric diaphragm pump, aiming to improve the accuracy and efficiency of drug dynamic signal generation in cell biology research; the device is composed of a dynamic chemical signal generation component and a microfluidic mixing chip, wherein the dynamic chemical signal generation component includes a piezoelectric diaphragm pump, a piezoelectric diaphragm pump control module and a piezoelectric diaphragm pump control module rack; the piezoelectric diaphragm pump can achieve precise flow control, and supports multiple pumps in parallel or series to increase flow; the microfluidic mixing chip is provided with multiple liquid inlets, and uniform mixing is achieved through multiple rows of mixing channels; the present invention can fully automatically control the pumping of liquids, reducing manual operations. The overall design of the present invention is compact and portable, suitable for desktop use, and provides convenience for cell experiments. The present invention can effectively generate single or complex dynamic chemical signals, accurately simulate the cell microenvironment, and is of great significance to the study of cell behavior.

Description

Technical Field

[0001] The present 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 cell biology research, administering drugs to cells to study their behavior is a common experimental approach. This approach helps scientists explore the effects of drugs on cellular physiological and pathological processes, including basic life processes such as cell proliferation, differentiation, apoptosis, and motility. Through these studies, scientists can uncover internal cellular signaling networks and their mechanisms of action, understand how cells respond to external signals, and how these responses influence cell fate and function.

[0003] Traditionally, this process requires manual operation, which is inefficient and easily affected by the operator's skill level. Currently, devices such as syringe pumps are used for precise drug addition, but existing injection devices are generally large in size. Conventional drug titration experiments achieve a dynamic drug signal by simply adding a certain amount of drug at regular intervals. This does not accurately control the concentration of the cell's environment, and the resulting signal is not a standard dynamic signal. Furthermore, whether performing manual titration or using a syringe pump, it is often difficult to precisely add trace amounts of drug. However, cells are extremely sensitive to their living environment, and large differences in the amount of drug added each time may affect cell behavior. Summary of the Invention

[0004] In view of the above 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 includes 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 is provided with a piezoelectric diaphragm pump;

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

[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 connection holes to communicate with the external liquid. The output end is connected to an output pipeline, and the output pipeline passes through another pipeline connection hole to connect to the microfluidic mixing chip. The control interface is controlled and 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 provided at one end of the carrier plate, and the output pipelines are respectively connected to the corresponding chip liquid inlets;

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

[0013] A liquid mixing channel, wherein multiple rows of disks 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 chip outlet, and the chip outlet is connected to an external cell culture device through a drainage line;

[0014] The observation chamber is arranged on the liquid mixing channel close to the liquid outlet of the chip.

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

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

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

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

[0019] A control board fixing hole is provided 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] Furthermore,

[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 operating method of the device for generating a dynamic chemical signal 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] An external computer control device is used to select and number the piezoelectric diaphragm pumps that need to be operated, and to send corresponding instructions to the corresponding piezoelectric diaphragm pumps, so that the input and output line pipes connected to the corresponding piezoelectric diaphragm pumps are filled with the above-mentioned liquid;

[0031] The operating 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 operates, passing the liquid into the merging cavity through the liquid inlet of the chip. After the liquid merges 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: In this mode, the input parameters 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 a sinusoidal output of the drug concentration in the liquid;

[0035] Liquid constant concentration output mode: In this mode, the input parameters 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, the cycle time, the duration within the waveform cycle, and the total operating time of the piezoelectric diaphragm pump, which can achieve a constant concentration pulse output of the drug in the liquid;

[0037] After inputting corresponding parameters to each piezoelectric diaphragm pump according to the selected liquid output mode, the total flow rate of all piezoelectric diaphragm pumps is input to the external computer control device, and the operation is performed to calculate 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, the pop-up window will be exited;

[0039] If so, an image of the flow rate of the selected piezoelectric diaphragm pump changing over time and an image of the target concentration of each drug changing 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 rate is driven by a piezoelectric diaphragm pump instead of a syringe or injection pump; taking water as an example, when driven by a voltage of 250V, the flow rate of a single pump ranges from 200 μL to 5000 μL; the device can also use piezoelectric diaphragm pumps in series or in parallel to increase the flow rate, which provides convenience for 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 the piezoelectric diaphragm pump to pump the liquid. Under the condition that the piezoelectric diaphragm pump voltage and the duty cycle of the PWM wave 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 piezoelectric diaphragm pump voltage, 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, thereby realizing the dynamic output of the drug in the liquid. The three typical modes are: sinusoidal output, constant concentration output, and pulse output mode. The dynamic output of any concentration of drug (chemical signal) 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 for generating dynamic chemical signals using a piezoelectric diaphragm pump of the present invention, the piezoelectric diaphragm pump can be fully automatically controlled to pump liquid, thereby reducing manual operation 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) The device for generating dynamic chemical signals using a piezoelectric diaphragm pump is compact 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, while the entire device is 130 mm long, 110 mm wide, and 110 mm high. The 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 the device for generating dynamic chemical signals using a piezoelectric diaphragm pump according to the present invention;

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

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

[0048] Figure 4 Schematic diagram of the piezoelectric diaphragm pump in the present invention;

[0049] Figure 5 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 over the total time in Case 2 of the present invention (the left one is isothiocyanate, the right one is rhodamine).

[0053] Among them, 1. Dynamic chemical signal generation 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 rack; 20. Chip liquid inlet; 21. Converging chamber; 22. Liquid mixing channel; 23. Observation chamber; 24. Chip liquid outlet. DETAILED DESCRIPTION

[0054] The following is combined with Figure 1 To the attached Figure 8 , a detailed description of the specific embodiments of the present invention is provided. 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," "inner," "outer," and the like, indicating directions or positional relationships, are based on the directions or positional relationships shown in the accompanying drawings and are intended only to facilitate the description of the present invention and simplify the description. They do not indicate or imply that the devices or components referred to must have a specific direction, be constructed, or operate in a specific direction. Therefore, they should not be construed as limiting the present invention.

[0055] The terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the quantity of the technical features being referred to. Thus, features specified as "first" or "second" may explicitly or implicitly include one or more of such features; and 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 includes 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 provided in the box-shaped housing 6. The piezoelectric diaphragm pump 3 is fixedly connected to the piezoelectric diaphragm pump slot 9 and is controlled and connected to the piezoelectric diaphragm pump control board 8. Two pipeline connection holes 12 are provided 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 to an input pipeline, which passes through a pipeline connection hole 12 and is in communication with the external liquid. The output end 16 is connected to an output pipeline, which passes through another pipeline connection hole 12 and is connected to the microfluidic mixing chip 2. The control interface 17 is connected to the piezoelectric diaphragm pump control board 8 for control.

[0061] like Figure 5 As shown, the microfluidic mixing chip 2 includes a carrier board, a chip liquid inlet 20, a confluence chamber 21, a liquid mixing channel 22 and an observation chamber 23; multiple chip liquid inlets 20 are arranged at one end of the carrier board, and the output pipelines are respectively connected to the chip liquid inlets 20; the confluence chamber 21 is arranged on the carrier board, and the confluence chamber 21 is connected to the output ports of the multiple chip liquid inlets 20; multiple rows of liquid mixing channels 22 are arranged on the carrier board, one end of the liquid mixing channel 22 is connected to the confluence chamber 21, and the other end is provided with a chip liquid outlet 24, and the chip liquid outlet 24 is connected to the external cell culture device through a discharge pipeline; the observation chamber 23 is arranged on the liquid mixing channel 22 near the chip liquid outlet 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 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 includes 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 external computer control device selects the number of piezoelectric diaphragm pumps 3 that need to be operated and numbers them, and sends corresponding instructions to the corresponding piezoelectric diaphragm pumps 3, so that the input and output line pipes connected to the corresponding piezoelectric diaphragm pumps 3 are filled with the above-mentioned liquid;

[0068] Setting the operating mode of each piezoelectric diaphragm pump 3 through an external computer control device: and inputting corresponding parameters to the piezoelectric diaphragm pump 3 according to the selected mode;

[0069] Each piezoelectric diaphragm pump 3 operates to pass the liquid into the merging chamber 21 through the chip liquid inlet 20. After the liquid is merged in the merging chamber 21, it enters the liquid mixing channel 22 for mixing. The evenly mixed liquid flows into the external cell culture device through the chip liquid outlet 24.

[0070] Setting the operating mode of the piezoelectric diaphragm pump 3 includes:

[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 operating 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 operating 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 rate of all piezoelectric diaphragm pumps 3 is input to the external computer control device, and the operation is performed to 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, the pop-up window will be exited;

[0076] 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.

[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 another 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 send an instruction to fill the piezoelectric diaphragm pump 3 and the input pipeline with the rhodamine solution, and the output end 16 of this 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 send an instruction to fill the piezoelectric diaphragm pump 3 and the input pipeline with deionized water, and the output end 16 of this 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 pin needles to prevent leakage;

[0079] Place the microfluidic mixing chip 2 under an inverted fluorescence microscope, set the fluorescence channel to TRITC, set the shooting time to five minutes, and the shooting interval to 500ms, and observe the instantaneous fluorescence intensity in 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, enter the initial drug concentration as 0.01 mg / ml, enter 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 buffer solution piezoelectric diaphragm pump control module 4, set the instantaneous maximum total flow rate to 10 ml / min, and click Start to start generating a single chemical signal;

[0081] The transient 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 the 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 the 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 the 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 pipelines; the input end 15 of the piezoelectric diaphragm pump 3 in the other 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] Place the microfluidic mixing chip 2 under an inverted fluorescence microscope, set the fluorescence channels to TRITC and FITC dual channels for simultaneous imaging, set the imaging time to 60 minutes, and the imaging interval to 5 seconds, and observe the instantaneous fluorescence intensity in 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 sine input mode, input the initial drug concentration as 0.01 mg / ml, input the sine maximum value as 4500 μL / Min, the minimum value as 1000 μL / Min, the initial phase as zero, the period as 300 s, and the total time as 60 minutes; select the piezoelectric diaphragm pump control module for inputting the fluorescein isothiocyanate solution 4, then select the mode of the piezoelectric diaphragm pump control module 4 for rhodamine solution as sine input mode, enter the initial drug concentration as 0.005 mg / ml, enter the sine 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 buffer solution, set the mode to constant frequency mode, set the flow rate to 5000 μL / Min, and click Start to start generating complex fluid chemical signals;

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

[0087] The specific models of the above electronic components are not particularly specified, and common 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 are only specific embodiments of the present invention and do 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 scope of protection of the present invention.

Claims

1. A device for generating dynamic chemical signals using a piezoelectric diaphragm pump, characterized in that: It includes 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 to 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 provided 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), wherein the input end (15) is connected to an input pipeline, and the input pipeline passes through one of the pipeline connection holes (12) to communicate with an external liquid, and the output end (16) is connected to an output pipeline, and the output pipeline passes through another of the pipeline connection holes (12) to connect to the microfluidic mixing chip (2), and the control interface (17) is control-connected to the piezoelectric diaphragm pump control board (8); The microfluidic mixing chip (2) comprises: carrier board; A plurality of chip liquid inlets (20) are provided at one end of the carrier plate, and the output pipelines are respectively connected to the chip liquid inlets (20) in a corresponding manner; A merging cavity (21) is provided 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), with multiple rows of discs arranged on the carrier plate, one end of the liquid mixing channel (22) is connected to the confluence cavity (21), and the other end is provided with a chip liquid outlet (24), and the chip liquid outlet (24) is connected to an external cell culture device through a drainage line; An observation chamber (23) is provided on the liquid mixing channel (22) near the liquid outlet (24) of the chip; The piezoelectric diaphragm pump control module frame (5) comprises: A partition fixing frame (19), wherein a plurality of the piezoelectric diaphragm pump control modules (4) are arranged at intervals on the partition fixing 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 provided below the partition fixing frame (19).

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

3. The device for generating dynamic chemical signals using a piezoelectric diaphragm pump according to claim 2, 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 (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 microfluidic mixing chip (2) is made of either PDMS polydimethylsiloxane or ABS plastic material.

4. A method for operating the device for generating dynamic chemical signals using a piezoelectric diaphragm pump according to claim 2, characterized in that: The steps include: 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; The number of piezoelectric diaphragm pumps (3) to be operated is selected and numbered by an external computer control device, and corresponding instructions are sent to the corresponding piezoelectric diaphragm pumps (3) so that the input line pipes and output line pipes connected to the corresponding piezoelectric diaphragm pumps (3) are filled with the above-mentioned liquid; Setting the operating mode of each piezoelectric diaphragm pump (3) through an external computer control device: and inputting corresponding parameters into the piezoelectric diaphragm pump (3) according to the selected mode; Each piezoelectric diaphragm pump (3) operates to pass the liquid into the merging cavity (21) through the chip liquid inlet (20). 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 chip liquid outlet (24).

5. The method for operating a device for generating a dynamic chemical signal using a piezoelectric diaphragm pump according to claim 4, characterized in that: The operating modes of the piezoelectric diaphragm pump (3) include: 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 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 operating 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).

6. The method for operating a device for generating dynamic chemical signals using a piezoelectric diaphragm pump according to claim 4, characterized in that: After inputting corresponding parameters to each piezoelectric diaphragm pump (3) according to the selected mode, the total flow rate of all the 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, the pop-up window will be exited; If so, an image of the flow rate of the selected piezoelectric diaphragm pump (3) changing with time and an image of the target concentration of each drug changing with time are popped up.

Images