Digital micro-fluidic chip temperature control system and method

Abstract

The invention discloses a digital micro-fluidic chip temperature control system, and the system comprises a heat conduction silica gel pad which is used for bearing a digital micro-fluidic chip; the array temperature sensor is used for sensing the temperature of the digital micro-fluidic chip on the heat-conducting silica gel pad; the heater is arranged below the heat conduction layer; the heat dissipation plate is arranged below the heater; the central processing unit is electrically connected with the heater, the temperature sensor and the heat dissipation plate, and the central processing unit enables the digital micro-fluidic chip to be at the optimal temperature in the amplification reaction circulation sub-process by adjusting the heating power of the heater and the rotating speed of the heat dissipation plate; the heater comprises a plurality of independent temperature control sub-areas which are used for being synchronously started in sequence after cycle times are spaced. By combining temperature control with an artificial intelligence method, real-time monitoring and prediction of the temperature of the reaction chamber are realized, and through real-time monitoring and prediction of the temperature of the reaction chamber and setting of the independent temperature control sub-regions, a plurality of amplification reaction cycles are started at intervals, so that the flexibility of amplification reaction setting is improved.

Description

[0001] The present invention belongs to the field of biomedical detection, and particularly relates to a temperature control system and method for a digital microfluidic chip. Background Art PCR thermal cycling includes three steps: denaturation, annealing, and extension. Denaturation means providing an environmental temperature of about 95°C to unwind the target double-stranded DNA into two single-stranded DNAs. Then, the environmental temperature is controlled to about 56°C to allow the primers to complementarily pair with the single-stranded DNA. Finally, the temperature is raised to about 72°C to enable the polymerase to complementarily pair the free nucleotides with the single-stranded DNA at an appropriate temperature and extend forward along the DNA strand.

[0003] Digital PCR technology is a nucleic acid molecule absolute quantification technology. It distributes a fluorescence quantitative PCR reaction system into a large number of tiny reactors, and there is only 0 - 1 nucleic acid of the target DNA sequence in each reactor for "single-molecule template PCR amplification". After the amplification is completed, the copy number of the target gene in the original sample is calculated through the number of positive reaction units (judged by the end-point fluorescence signal) and statistical methods. Digital PCR can perform accurate absolute quantification detection without relying on control samples and standard curves. In addition, since digital PCR only needs to judge two amplification states of "yes / no" when interpreting the results, it does not need to know the initial concentration of the target gene in advance like real-time quantitative PCR technology.

[0004] The digital PCR amplification stage is similar to PCR thermal cycling. During the amplification process, the requirements for temperature in the three stages of denaturation, annealing, and extension are different. Therefore, the temperature module of the digital PCR instrument should precisely and quickly control the temperature.

[0005] Domestic manufacturers produce digital PCR instruments, but they generally have the disadvantages of long heating time and slow reaction speed. Summary of the Invention

[0006] In view of the deficiencies of the prior art, the technical solutions adopted by the present invention to solve the above technical problems are as follows: A temperature control system for a digital microfluidic chip, the system includes: A thermally conductive silicone pad for carrying the digital microfluidic chip; An array temperature sensor is arranged inside the silicone pad for sensing the temperature of the digital microfluidic chip on the thermally conductive silicone pad; A thermally conductive layer is arranged below the thermally conductive silicone pad, and the thermally conductive layer is composed of stacked copper sheets and aluminum sheets; A heater is arranged below the thermally conductive layer; A heat dissipation plate is arranged below the heater; the heat dissipation plate is composed of materials with heat dissipation functions. The heat dissipation plate can use air circulation for heat dissipation or condensing liquid for heat dissipation. A motor is arranged inside the heat dissipation plate, and the rotation of the blades enables the acceleration of air circulation or condensing liquid circulation. The central processing unit is electrically connected to the heater, temperature sensor, and heat dissipation plate. The central processing unit makes the digital microfluidic chip reach the optimal temperature during the amplification reaction cycle sub-process by adjusting the heating power of the heater and the rotation speed of the motor inside the heat dissipation plate; The heater includes multiple independently temperature-controlled sub-regions, which are used to start synchronously in sequence after a certain number of cycle intervals.

[0007] Further, the central processing unit includes a single-chip microcomputer, DSP, ARM, FPGA, PLC, Further, the central processing unit includes a peripheral control circuit, and the peripheral control circuit includes: An AD / DC circuit is used to convert the 220V voltage into 48V and 12V DC voltages to supply power to the central processing unit and the heat dissipation plate, A heating control circuit generates a PWM signal through the central processing unit and transmits it to the MOSFET through an optocoupler. When the PWM is at a high level, the MOSFET conducts and the heater is turned on. The power of the heater is changed by adjusting the PWM duty cycle; the heater includes multiple independently operating sub-regions; A refrigeration control circuit generates a PWM signal through the central processing unit to adjust the rotation speed of the motor inside the heat dissipation plate; A temperature acquisition circuit includes thin-film thermal resistors arrayed in the independently temperature-controlled sub-regions, which convert the temperature value into a resistance value; Further, the central processing unit makes the digital microfluidic chip reach the optimal temperature during the amplification reaction cycle sub-process by adjusting the heating power of the heater and the rotation speed of the motor inside the heat dissipation plate, including the central processing unit adjusting the heater through a fuzzy RBF neural network model. The steps are as follows: S1. Determine the temperature control range and temperature control duration of the digital microfluidic chip; S2. Model the temperature control system of the digital microfluidic chip, including heat transfer, heat loss, and heat control units; S3. Define fuzzy sets and fuzzy rules, and the fuzzy sets are used to represent temperature and temperature difference variables, S4. Construct an RBF neural network model, including an input layer, a fuzzyfication layer, a fuzzy inference layer, and an output layer, S5. Train the fuzzy RBF network according to the simulation data to optimize the RBF center, width, and weight; S6. According to the fuzzy RBF control algorithm, convert the network output into a PWM signal to drive the heater.

[0008] Further, the temperature control method further includes: the temperature control algorithm can be PID control, fuzzy control, neural network control, adaptive control, fuzzy PID control, adaptive PID control, or fuzzy adaptive PID control; Further, the heat conduction layer is composed of stacked copper sheets and aluminum sheets.

[0009] Further, the amplification reaction cycle sub-process includes a denaturation process, an annealing process, and an extension process.

[0010] Generally, for the PCR temperature cycle process in the PCR reaction, the process is as follows: First, the PCR temperature cycle parameters need to be preset according to the characteristics of the selected sample DNA. Then, start the temperature cycle device. Input the temperature value in the PCR temperature unit collected in real time during the PCR temperature cycle process to the adopted controller. Then, adjust the power of the actuator according to the formulated control law to achieve the PCR temperature cycle process. And in order to facilitate subsequent analysis, the data should be sent to the human-machine interaction interface in real time.

[0011] The parameter settings of the PCR reaction conditions and their impacts are as described below. (1) Settings of PCR reaction temperature and time: The temperature and time settings for each temperature zone of PCR are as follows: 1) Denaturation: During the PCR reaction, if the denaturation temperature is too low, it will have a great impact on the final DNA amplification result. Generally, the denaturation temperature of the DNA template is maintained at 93°C - 94°C for 1 minute. If it is lower than 93°C, the time needs to be extended, but the temperature cannot be too high because the high-temperature environment affects the enzyme activity. If this step cannot completely denature the target gene template or PCR product, it will lead to the failure of the PCR reaction.

[0012] 2) Annealing: The annealing temperature is an important factor affecting specificity. After denaturation, the temperature is rapidly cooled to 40°C - 60°C, which can cause the primer and the template to bind. Generally, the ideal starting point for the annealing temperature is around 55°C (30 - 60S).

[0013] 3) Extension: The extension temperature of the reaction is generally selected between 70 - 75°C, and the commonly used temperature is 72°C (generally 1 minute). Too high an extension temperature is not conducive to the binding of the primer and the template. Too long an extension time will lead to the appearance of non-specific amplification bands. For the amplification of low-concentration templates, the extension time should be slightly longer.

[0014] (2) Number of cycles: The number of cycles determines the degree of PCR amplification. The number of PCR cycles (generally 30 - 60 times) mainly depends on the concentration of the template DNA. If the set number of cycles is more, the amount of non-specific products will also increase.

[0015] Furthermore, the optimal temperature and duration of the denaturation process are set to 93°C for 1 min; the optimal temperature and duration of the annealing process are set to 55°C for 40 s, the optimal temperature and duration of the extension process are set to 70°C for 1 min, and the number of cycles is 50 times.

[0016] Furthermore, the heater includes multiple independently temperature-controlled sub-regions, which are used to start synchronously in sequence after a certain number of cycle intervals. Among them, the concentration of template DNA in the second temperature-controlled sub-region is adjusted according to the concentration of the initial template DNA and the cyclic amplification result of the first temperature-controlled sub-region.

[0017] A method for controlling the temperature of a digital microfluidic chip includes: S1. The central processing unit adjusts the heating power of the heater and the rotation speed of the motor in the heat sink through fuzzy PID control so that the digital microfluidic chip is at the optimal temperature during the cyclic sub-process of the amplification reaction; including: S11. Set the preset temperature of the heater and the preset rotation speed of the motor in the heat sink, turn on the heater according to the preset temperature, and start the motor in the heat sink according to the preset rotation speed of the motor in the heat sink; S12. Collect the ambient temperature, input the digital PCR temperature into the LSTM neural network model to obtain the predicted digital PCR temperature; S13. Input the predicted digital PCR temperature, the output power of the heater, and the rotation speed of the motor in the heat sink into the fuzzy PID control system, S14. The fuzzy PID control system adjusts the output power of the heater and the rotation speed of the motor in the heat sink according to the difference between the preset temperature and the digital PCR temperature until the difference between the preset temperature and the digital PCR temperature is less than the first threshold; Among them, the LSTM model includes an input gate, a cell state, and an output gate, The input gate receives the ambient temperature, digital PCR temperature, output power of the heater, and rotation speed of the motor in the heat sink collected by the temperature sensor array at the current moment, The output gate outputs the predicted digital PCR temperature, the heating power of the heater, and the rotation speed of the motor in the heat sink; When the absolute value of the temperature difference is greater than the second threshold, the heating power of the heater and the rotation speed of the motor in the heat sink are adjusted simultaneously, otherwise only the heating power of the heater or the rotation speed of the motor in the heat sink is adjusted; S2. Adjust the concentration of template DNA in the second temperature-controlled sub-region according to the concentration of the initial template DNA and the cyclic amplification result of the first temperature-controlled sub-region, including: S21. Collect the concentration of the initial template DNA, the temperature curve, the number of cycles, and the amplification result of the first temperature-controlled sub-region; S22. Compare the deviation values of the temperature curve, the number of cycles, and the amplification result of the first temperature-controlled sub-region with the standard temperature curve, the number of cycles, and the amplification result; S23. Adjust the concentration and / or number of cycles of the second temperature-controlled sub-region template DNA according to the deviation value; The temperature curve deviation is the area difference.

[0018] A computer-readable storage medium stores a computer program, characterized in that a processor executes the computer program to implement a digital microfluidic chip temperature control method.

[0019] A terminal device includes a memory, a processor, and a computer program stored in the memory and executable on the processor, characterized in that the processor executes the computer program to implement a digital microfluidic chip temperature control method.

[0020] The beneficial effects of the present invention are as follows: 1) Through the real-time monitoring and prediction of the reaction chamber temperature, the system can automatically adjust the temperature of the heating element, thereby achieving precise control of the reaction temperature. Through the simulation and prediction of the reaction process, the reaction conditions and the concentration of reactants can be automatically adjusted, thereby improving the efficiency and accuracy of the reaction.

[0021] 2) By setting independent temperature-controlled sub-regions, multiple amplification reaction cycles are started at intervals, improving the flexibility of the amplification reaction settings.

[0022] The above description is only an overview of the technical solution of the present invention. In order to be able to more clearly understand the technical means of the present invention, it can be implemented according to the content of the specification. And in order to make the above description and other purposes, features and advantages of the present invention more obvious and understandable, specific preferred embodiments are given and described in detail as follows. Brief Description of the Drawings Figure 1 It is a structural diagram of a digital microfluidic chip temperature control system; Figure 2 It is a logic diagram of a digital microfluidic chip temperature control method; Figure 3 It is a temperature response diagram of a digital microfluidic chip temperature control system. Detailed Embodiments The exemplary embodiments of the present disclosure will be described in more detail below with reference to the drawings. Although the exemplary embodiments of the present disclosure are shown in the drawings, it should be understood that the present disclosure can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided so that the present disclosure can be more thoroughly understood and the scope of the present disclosure can be fully conveyed to those skilled in the art.

[0025] In the description of the present invention, unless otherwise clearly defined and limited, terms such as "installation", "connection", "coupling", "fixation" should be understood in a broad sense. For example, it can be a connection, a detachable connection, or integrated; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium, and it can be the communication inside two components or the interaction relationship between two components. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0026] Embodiment 1 A temperature control system for a digital microfluidic chip, the system comprising a thermally conductive silicone pad, an array temperature sensor, a heat conducting layer, a heating layer, and a heat sink.

[0027] The thermally conductive silicone pad is used to carry the digital microfluidic chip; the array temperature sensor is disposed inside the silicone pad and is used to sense the temperature of the digital microfluidic chip on the thermally conductive silicone pad; the heat conducting layer can be a copper sheet heat conducting layer, and the copper sheet heat conducting layer is disposed below the thermally conductive silicone pad, and the copper sheet heat conducting layer is composed of stacked copper sheets and aluminum sheets.

[0028] A heater is disposed below the heat conducting layer. The heater includes a plurality of independently temperature-controlled sub-regions, which are used to be sequentially and synchronously started after a certain number of cycle intervals. Among them, the concentration of the template DNA in the second temperature control sub-region is adjusted according to the concentration of the initial template DNA and the cycle amplification result of the first temperature control sub-region. For example, the first temperature control sub-region causes the first digital PCR to perform cycle amplification according to preset parameters. After 20 cycles of amplification, the concentration of the template DNA after cycle amplification in the first temperature control sub-region is detected. If the concentration meets the expectation, the subsequent cycle amplification continues; otherwise, after modifying the preset parameters of the second temperature control sub-region, such as correcting the temperature curve or adjusting the concentration of the template DNA and then performing cycle amplification. The above operations form a comparison before and after modification to form a matching correction result pair. At the same time, the above temperature control system may further include a third temperature control sub-region. The preset initial parameters of the third temperature control sub-region are different from those of the first temperature control sub-region, which can be the temperature curve, temperature control logic, and concentration of the template DNA, and perform cycle amplification synchronously, directly performing the cycle amplification process of multiple independent digital PCRs, saving the repeatedly set procedures; further, multiple digital PCRs can be placed in the temperature control sub-region.

[0029] The heat sink is disposed below the heater. The heat sink is composed of materials with heat dissipation functions. The heat sink can use air circulation for heat dissipation or condensing liquid for heat dissipation. A motor is arranged inside the heat sink, and the rotation of the blades accelerates the air circulation or the circulation of the condensing liquid. The central processing unit is electrically connected to the heater, the temperature sensor, and the heat sink. The central processing unit adjusts the heating power of the heater and the rotation speed of the motor inside the heat sink to keep the digital microfluidic chip at the optimal temperature during the amplification reaction cycle sub-process. The amplification reaction cycle sub-process includes a denaturation process, an annealing process, and an extension process. The optimal temperature and duration of the denaturation process are set to 93°C for 1 minute; the optimal temperature and duration of the annealing process are set to 55°C for 40 seconds, and the optimal temperature and duration of the extension process are set to 70°C for 1 minute.

[0030] The central processing unit includes a peripheral control circuit. Among them, the peripheral control circuit includes: The AD / DC circuit is used to convert the 220V voltage into 48V and 12V DC voltages to supply power to the central processing unit and the heat sink. The heating control circuit generates a PWM signal through the central processing unit and transmits it to the MOSFET through an optocoupler. When the PWM is at a high level, the MOSFET conducts, and the heater is turned on. The power of the heater is changed by adjusting the PWM duty cycle. The heater includes multiple sub-regions that work independently. The refrigeration control circuit generates a PWM signal through the central processing unit to adjust the rotation speed of the motor inside the heat sink. The temperature acquisition circuit includes thin-film thermal resistors distributed in an array in the independently temperature-controlled sub-regions, which convert the temperature value into a resistance value. Furthermore, the central processing unit adjusts the heating power of the heater and the rotation speed of the motor inside the heat sink to keep the digital microfluidic chip at the optimal temperature during the amplification reaction cycle sub-process, including the central processing unit adjusting the heater through a fuzzy RBF neural network model. The steps are as follows: S1. Determine the temperature control range, accuracy, and response time of the digital microfluidic chip. S2. Model the temperature control system of the digital microfluidic chip, including heat transfer, heat loss, and heat control units. S3. Define fuzzy sets and fuzzy rules. The fuzzy sets are used to represent temperature and temperature difference variables. S4. Construct an RBF neural network model, including an input layer, a fuzzification layer, a fuzzy inference layer, and an output layer. S5. Train the fuzzy RBF network according to the simulation data to optimize the RBF center, width, and weight. S6. According to the fuzzy RBF control algorithm, convert the network output into a PWM signal to drive the heater.

[0031] Example 2 A method for controlling the temperature of a digital microfluidic chip, comprising: S1. The central processing unit adjusts the heating power of the heater and the rotational speed of the motor in the heat sink through fuzzy PID control so that the digital microfluidic chip is at the optimal temperature during the amplification reaction cycle sub-process; including: S11. Set the preset temperature of the heater and the preset rotational speed of the motor in the heat sink, turn on the heater according to the preset temperature, and start the motor in the heat sink according to the preset rotational speed of the motor in the heat sink; S12. Collect the ambient temperature, input the digital PCR temperature into the LSTM neural network model to obtain the predicted digital PCR temperature; S13. Input the predicted digital PCR temperature, the output power of the heater, and the rotational speed of the motor in the heat sink into the fuzzy PID control system, S14. The fuzzy PID control system adjusts the output power of the heater and the rotational speed of the motor in the heat sink according to the difference between the preset temperature and the digital PCR temperature until the difference between the preset temperature and the digital PCR temperature is less than the first threshold; Wherein, the LSTM model includes an input gate, a cell state, and an output gate, The input gate receives the ambient temperature, digital PCR temperature, and output power of the heater, and the rotational speed of the motor in the heat sink collected by the temperature sensor array at the current moment, The output gate outputs the predicted digital PCR temperature, the heating power of the heater, and the rotational speed of the motor in the heat sink; When the absolute value of the temperature difference is greater than the second threshold, the heating power of the heater and the rotational speed of the motor in the heat sink are adjusted simultaneously, otherwise only the heating power of the heater or the rotational speed of the motor in the heat sink is adjusted; S2. Adjust the concentration of the template DNA in the second temperature control sub-region according to the concentration of the initial template DNA and the amplification result of the first temperature control sub-region cycle amplification, including: S21. Collect the concentration of the initial template DNA, the temperature curve, the number of cycles, and the amplification result of the first temperature control sub-region; S22. Compare the deviation values of the temperature curve, the number of cycles, and the amplification result of the first temperature control sub-region with the standard temperature curve, the number of cycles, and the amplification result; S23. Adjust the concentration and / or the number of cycles of the template DNA in the second temperature control sub-region according to the deviation value; The temperature curve deviation is the area difference.

[0032] S24. Set the preset initial parameters of the third temperature control sub-region to be different from those of the first temperature control sub-region, synchronously perform cycle amplification, and compare the amplification results of the first temperature control sub-region and the third temperature control sub-region.

[0033] To ensure that the temperature change of the system meets the requirements of the PCR reaction, the following settings are made: heating from room temperature to 93°C and maintaining a constant temperature of 93°C; cooling from 93°C to 55°C and maintaining a constant temperature of 55°C; heating from 55°C to 72°C and maintaining a constant temperature of 72°C. For details, see Figure 3 。After placing the test tube containing the reaction simulation reagent into the sample pool, insert the Pt100 sensor into the test tube. Use the ADC module to sample the output voltage of the temperature sensing circuit. The sampling mode is set to sequential sampling with a frequency of 200 Hz. By plotting the temperature curve, perform 10 groups of experiments and take the average value. The sample pool can cycle within the set temperature range, maintain a constant temperature, with a heating and cooling rate of 4.2°C / s and an accuracy of ±0.2°C.

[0034] Experimental result table A computer-readable storage medium stores a computer program, characterized in that a processor executes the computer program to implement a digital microfluidic chip temperature control method.

[0035] The beneficial effects of the present invention are as follows: 1) Through the real-time monitoring and prediction of the reaction chamber temperature, the system can automatically adjust the temperature of the heating element, thereby achieving precise control of the reaction temperature. Through the simulation and prediction of the reaction process, the reaction conditions and the concentration of reactants can be automatically adjusted, thereby improving the efficiency and accuracy of the reaction.

[0036] 2) By setting up independent temperature control sub-regions, multiple amplification reaction cycles are started at intervals, improving the flexibility of the amplification reaction settings.

Claims

1. A digital microfluidic chip temperature control system, characterized in that: The system includes: Thermally conductive silicone pads for carrying digital microfluidic chips; An array temperature sensor is arranged inside the silicone pad and is used to sense the temperature of the digital microfluidic chip on the thermally conductive silicone pad; The heat-conducting layer is arranged below the heat-conducting silicone pad; A heater is disposed below the heat conductive layer; A heat dissipation plate is arranged below the heater; The central processing unit is electrically connected to the heater, the temperature sensor, and the heat sink. The central processing unit adjusts the heating power of the heater and the speed of the motor in the heat sink so that the digital microfluidic chip is at an optimal temperature during the amplification reaction cycle subprocess. The heater comprises a plurality of independently temperature-controlled sub-areas, which are used to start synchronously in sequence after a certain number of cycles.

2. The digital microfluidic chip temperature control system according to claim 1, characterized in that: The central processing unit includes a peripheral control circuit, and the peripheral control circuit includes: AD / DC circuit, used to convert 220V voltage into 48V and 12V DC voltage to power the CPU and heat sink. The heating control circuit generates a PWM signal through the central processor and transmits it to the MOSFET through the optocoupler. When the PWM is at a high level, the MOSFET is turned on and the heater is turned on. The power of the heater is changed by adjusting the PWM duty cycle. The heater includes multiple sub-areas that work independently. The cooling control circuit generates a PWM signal through the central processor to adjust the motor speed in the heat sink; The temperature acquisition circuit includes thin-film thermal resistors distributed in arrays in independently temperature-controlled sub-areas, which convert temperature values ​​into resistance values.

3. The digital microfluidic chip temperature control system according to claim 1, characterized in that: The central processor adjusts the heating power of the heater and the speed of the motor in the heat sink so that the digital microfluidic chip is at the optimal temperature in the amplification reaction cycle sub-process, including the central processor adjusting the heater by performing a fuzzy RBF neural network model, the steps are as follows: S1. Determine the temperature range, accuracy and response time of the digital microfluidic chip temperature control; S2. Model the temperature control system of the digital microfluidic chip, including heat transfer, heat loss and thermal control unit; S3, define fuzzy sets and fuzzy rules, the fuzzy sets are used to represent temperature and temperature difference variables, S4. Construct the RBF neural network model, including input layer, fuzzification layer, fuzzy reasoning layer and output layer. S5, training the fuzzy RBF network according to the simulated data, and optimizing the RBF center, width and weight; S6. According to the fuzzy RBF control algorithm, the network output is converted into a PWM signal for driving the heater.

4. The digital microfluidic chip temperature control system according to claim 1, characterized in that: The heat conducting layer is composed of stacked copper sheets and aluminum sheets.

5. The digital microfluidic chip temperature control system according to claim 1, characterized in that: The amplification reaction cycle sub-processes include denaturation process, annealing process, and extension process.

6. The digital microfluidic chip temperature control system according to claim 5, characterized in that: The optimal temperature and duration of the denaturation process were set at 93°C for 1 min; the optimal temperature and duration of the annealing process were set at 55°C for 40S, and the optimal temperature and duration of the extension process were set at 70°C for 1 min.

7. The digital microfluidic chip temperature control system according to claim 1, characterized in that: The heater includes a plurality of independently temperature-controlled sub-areas, which are used to start synchronously in sequence after an interval of cycles, wherein the concentration of the template DNA in the second temperature-controlled sub-area is adjusted according to the concentration of the initial template DNA and the cyclic amplification result of the first temperature-controlled sub-area.

8. A digital microfluidic chip temperature control method, applied to the digital microfluidic chip temperature control system as claimed in claim 1, characterized in that: S1. The central processing unit adjusts the heating power of the heater and the motor speed in the heat sink through fuzzy PID control so that the digital microfluidic chip is at the optimal temperature during the amplification reaction cycle sub-process; including: S11, setting a preset temperature of the heater and a preset speed of the motor in the heat sink, turning on the heater according to the preset temperature, and starting the motor in the heat sink according to the preset speed of the motor in the heat sink; S12, collecting the ambient temperature, and inputting the digital PCR temperature into the LSTM neural network model to obtain the predicted digital PCR temperature; S13, input the predicted digital PCR temperature, heater output power, and motor speed in the heat sink into the fuzzy PID control system, S14, the fuzzy PID control system adjusts the heater output power and the motor speed in the heat sink according to the difference between the preset temperature and the digital PCR temperature, until the difference between the preset temperature and the digital PCR temperature is less than a first threshold; Among them, the LSTM model includes input gate, cell state and output gate. The input gate receives the temperature sensor array to collect the current ambient temperature, digital PCR temperature and heater output power, and the motor speed in the heat sink. The output gate outputs the predicted digital PCR temperature, heater heating power, and motor speed in the heat sink; When the absolute value of the temperature difference is greater than the second threshold, the heating power of the heater and the speed of the motor in the heat sink are adjusted simultaneously, otherwise only the heating power of the heater or the speed of the motor in the heat sink is adjusted; S2, adjusting the concentration of the template DNA in the second temperature-controlled sub-region according to the concentration of the initial template DNA and the cyclic amplification result of the first temperature-controlled sub-region, including: S21, collecting the concentration of the initial template DNA, the temperature curve of the first temperature control sub-region, the number of cycles, and the amplification results; S22, comparing the temperature curve, cycle number and amplification result of the first temperature control sub-region with the deviation value of the standard temperature curve, cycle number and amplification result; S23, adjusting the concentration and / or cycle number of the template DNA in the second temperature control sub-region according to the deviation value; The temperature curve deviation is an area difference.

9. A computer-readable storage medium storing a computer program, characterized in that: The processor executes the computer program to implement the digital microfluidic chip temperature control method as claimed in claim 8.

10. A terminal device, comprising a memory, a processor, and a computer program stored in the memory and executable on the processor, characterized in that: The processor executes the computer program to implement the digital microfluidic chip temperature control method as claimed in claim 8.

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