Zebra fish pain response testing device for Chinese herbal medicine breeding

Abstract

The invention discloses a zebra fish pain response testing device for Chinese herbal medicine breeding, which comprises a plurality of branch water tanks, the plurality of branch water tanks are connected through a G selection pool to form a six-way water tank, and the plurality of branch water tanks are communicated with the G selection pool; the tail end of each branch water tank is an electric shock end for the zebra fish to receive electrical pain stimulation, the electric shock end is provided with an electrode plate for providing electrical stimulation for the zebra fish, an inlet of the electric shock end is provided with an infrared correlation trigger area for detecting whether the zebra fish passes through or not, and the electrode plate is connected with an electric shock module for providing voltage for the electrode plate. The technology has a multi-channel parallel test capability and an automatic electrical stimulation triggering function, can shorten an experimental period, reduces invalid stimulation, provides stable and reliable data support for pain mechanism research and drug screening, and improves research efficiency and scientificity.

Description

Technical Field

[0001] This technology relates to the field of pain response devices, specifically a zebrafish pain response testing device for the selection and breeding of traditional Chinese medicine. Background Technology

[0002] Wild chrysanthemum, a traditional Chinese medicine, contains components with analgesic and anti-inflammatory activities (such as flavonoids and volatile oils). Different varieties of wild chrysanthemum have different types and contents of active ingredients, resulting in varying degrees of analgesic effects.

[0003] Breeding experts frequently use pain response testing devices to screen for strains with the strongest analgesic effects for further breeding. The specific method is as follows: Extracts from different varieties of wild chrysanthemum are applied to zebrafish before electrical stimulation using the pain response testing device. After electrical stimulation, the intensity of the zebrafish's pain response (e.g., whether the zebrafish's movement speed and distance significantly decreased), the duration of the pain response (e.g., how long did the painful writhing behavior last), and changes in behavioral patterns (e.g., whether the frequency and duration of abnormal behaviors (e.g., circling, body bending) decreased) are monitored and recorded. If the zebrafish in the wild chrysanthemum extract treatment group showed a significantly weaker pain response than the control group (untreated, i.e., no wild chrysanthemum extract applied) or other variety treatment groups, it indicates that the wild chrysanthemum variety has a better analgesic effect, i.e., superior quality. Through this rapid and objective bioactivity evaluation method, breeding experts can efficiently screen for strains with the strongest analgesic effects from a large number of hybrid or naturally mutated wild chrysanthemum offspring for further breeding. This avoids the bottleneck of traditionally requiring years of chemical composition analysis or mammalian efficacy testing, and enables rapid breeding based on efficacy.

[0004] Zebrafish, as a model organism, has a relatively clear genome that is highly similar to that of humans, making it widely and deeply used in behavioral research. Its pain transmission pathways and molecular mechanisms are highly conserved in mammals, effectively mimicking human pain responses. Furthermore, zebrafish are characterized by rapid reproduction, small size, and low rearing costs, facilitating high-throughput experiments. Their transparent larval stage allows for direct observation of neural activity, making them an ideal model for pain mechanism research and drug screening.

[0005] Traditional pain response testing devices for providing electrical stimulation are mostly single-channel designs, which cannot perform multiple parallel experiments simultaneously, resulting in low experimental efficiency. Alternatively, the stimulation module may lack precision, with a narrow range of electrical stimulation parameter control and poor stability, easily leading to fluctuations in experimental data. Or, the device structure may lack standardization, with inconsistent channel sizes and connection methods, affecting experimental repeatability. Furthermore, traditional pain response testing devices are mostly manually triggered or timed electrical stimulation, which cannot be linked to the zebrafish's position, further limiting experimental efficiency.

[0006] Therefore, a novel zebrafish pain response testing device with multi-channel parallel testing capability, high-precision stimulation regulation function, standardized structural design and automatic triggering electrical stimulation function is urgently needed for the selection and breeding of traditional Chinese medicine. This device is of great significance for the study of pain nerve mechanisms, screening of analgesic drugs, assessment of environmental neurotoxicity, and improvement of experimental accuracy and throughput. Summary of the Invention

[0007] The technical problem to be solved by this technology is to provide a zebrafish pain response testing device for the breeding of traditional Chinese medicine, which addresses the shortcomings of the existing technology. This zebrafish pain response testing device for the breeding of traditional Chinese medicine has multi-channel parallel testing capability and automatic triggering electrical stimulation function, which can shorten the experimental cycle, reduce ineffective stimulation, provide stable and reliable data support for pain mechanism research and drug screening, and improve research efficiency and scientificity.

[0008] To achieve the above technical objectives, the technical solution adopted in this invention is as follows:

[0009] A zebrafish pain response testing device for the selection and breeding of traditional Chinese medicine includes multiple branch tanks, which are connected by a G-selection pool to form a six-way tank, and all the branch tanks are connected to the G-selection pool.

[0010] Each branch tank has an electric shock end at its end, where zebrafish receive electric pain stimulation. The electric shock end is equipped with electrode plates for providing electric stimulation to the zebrafish. The entrance of the electric shock end is equipped with an infrared beam triggering area for detecting whether a zebrafish has passed by. The electrode plates are connected to an electric shock module for providing voltage to the electrode plates.

[0011] As a further improvement to this technology, there are six branch water tanks, which are respectively designated as branch water tank A, branch water tank B, branch water tank C, branch water tank D, branch water tank E and branch water tank F. Branch water tanks A, B, C, D, E and F are connected to the regular hexagonal G selection pool by bolts, nuts and washers.

[0012] As a further improvement to this technology, each branch water tank is provided with a slot for placing a baffle 5mm away from the G selection pool.

[0013] As a further improvement to this technology, each branch water tank includes a channel section and an electric shock end, which are interconnected. The end of the channel section is the inlet of the electric shock end, and the bottom of the electric shock end is lower than the bottom of the channel section.

[0014] As a further improvement to this technology, the electric shock module includes a transformer module, a rectifier and filter circuit, an operational amplifier and bias circuit, a main controller, a power supply auxiliary module, and an OLED display module;

[0015] The input terminal of the transformer module is connected to the mains power, the output terminal of the transformer module is connected to the input terminal of the rectifier and filter circuit, the output terminal of the rectifier and filter circuit is connected to the power supply auxiliary module, the power supply auxiliary module is connected to the operational amplifier and bias circuit, the main controller and the OLED display module respectively, the main controller is connected to the input terminal of the operational amplifier and bias circuit, the output terminal of the operational amplifier and bias circuit is connected to both the current sampling circuit and the electrode plate, the output terminal of the current sampling circuit is connected to the input terminal of the main controller, and the output terminal of the main controller is connected to the OLED display module.

[0016] As a further improvement to this technology, the infrared beam triggering area includes an infrared beam sensor. Windows facing each other are opened on both sides of the electric shock end inlet. Transparent glass is fixedly connected to each of the two windows. The outer side of one transparent glass is equipped with the transmitter of the infrared beam sensor, and the outer side of the other transparent glass is equipped with the receiver of the infrared beam sensor. The transmitter of the infrared beam sensor is connected to the power auxiliary module, and the receiver of the infrared beam sensor is connected to the main controller. The infrared beam sensor is used to detect whether a zebrafish is passing by.

[0017] As a further improvement to this technology, the transformer module receives mains power and converts it to 15V AC voltage. The rectifier and filter circuit converts the 15V AC voltage to 15V DC voltage. The power supply auxiliary module uses an LM2596 step-down circuit to convert the 15V DC voltage to 12V and 5V. The 12V voltage is used as input to the operational amplifier and bias circuit, and the 5V voltage is used as input to the main controller, OLED display module, and infrared beam sensor. The operational amplifier and bias circuit includes a bias voltage circuit and an operational amplifier circuit connected in sequence. The bias voltage circuit includes an amplifier OPA197, and the operational amplifier circuit includes an amplifier HTCOP07. The circuit is used to differentially amplify and power amplify the received voltage and output a drive voltage to one electrode plate. The other electrode plate is connected to the ground wire. The two electrode plates are respectively connected to the inner wall of the electric shock end. The current sampling circuit includes a current detection chip INA282. The current sampling circuit is used to collect the current passing through the electrode plate and feed it back to the main controller. The main controller includes a microcontroller STM32CBT6. The main controller is used to receive and analyze the signal sent by the infrared beam sensor. It is also used to send signals to the operational amplifier and bias circuit and the OLED display module respectively. The OLED display module is used to display the information collected by the current sampling circuit and the working time of the electrode plate according to the control instructions of the main controller.

[0018] As a further improvement to this technology, a tracking and recording device is provided above the six-way water tank.

[0019] As a further improvement to this technology, the tracking and recording device is connected to a six-way water tank via a tripod-shaped mounting bracket.

[0020] As a further improvement to this technology, the six-way water tank is made of transparent but opaque acrylic material.

[0021] The beneficial effects of this technology are:

[0022] This technology features six branch water tanks, which are connected by a G-selection pool to form a six-way water tank. Each branch water tank can provide electrical stimulation, i.e., to elicit a pain response. It has multi-channel parallel testing capability, which improves testing efficiency.

[0023] This technology uses an electric shock module to provide voltage to the electrode pads, thereby stimulating zebrafish to produce a pain response. The electric shock module includes a transformer module, a rectifier and filter circuit, an operational amplifier and bias circuit, a main controller, a power supply auxiliary module, and an OLED display module. Through the above modules, circuits, and main controller, a continuously adjustable voltage of 0~9.9V can be provided to the electrode pads, providing high-precision stimulation control and improving testing accuracy.

[0024] This technology includes multiple interconnected branch water tanks. The multiple branch water tanks and the G selection pool are connected by bolts, nuts and washers to form a six-way water tank. It adopts a standardized structure and a unified connection method, which is convenient for assembly and disassembly and allows for repeatable testing experiments.

[0025] This technology also includes an infrared beam triggering zone to detect whether a zebrafish is passing by and sends a signal to the main controller as a condition for triggering an electric shock. This enables automatic electric shock triggering linked to the zebrafish's position, providing an automatic electric stimulation triggering function and improving experimental efficiency.

[0026] In summary, this technology, through its multi-channel parallel testing capabilities, high-precision stimulation modulation function, standardized structural design, and automated stimulation triggering function, can shorten the experimental cycle, ensure experimental repeatability, reduce ineffective stimulation, provide stable and reliable data support for pain mechanism research and drug screening, and improve research efficiency and scientific rigor. Attached Figure Description

[0027] Figure 1 This is a three-dimensional schematic diagram of the technology.

[0028] Figure 2 This is a three-dimensional schematic diagram of the technology.

[0029] Figure 3 This is a top view of the technology.

[0030] Figure 4 This is a schematic diagram of the G-selection pool structure of this technology.

[0031] Figure 5 This is a schematic diagram of the circuit framework of the electric shock module of this technology.

[0032] Figure 6 This is a partial circuit diagram of the electric shock module of this technology.

[0033] Figure 7 This is a flowchart of the electric shock module software of this technology. Detailed Implementation

[0034] The specific implementation methods of this technology will be further described below with reference to the accompanying drawings:

[0035] like Figure 1-3 As shown, a zebrafish pain response testing device for the selection and breeding of traditional Chinese medicine includes multiple branch tanks. These branch tanks are connected by a regular hexagonal selection pool 8 to form a six-way tank. The six branch tanks are designated as branch tank A1, branch tank B2, branch tank C3, branch tank D4, branch tank E5, and branch tank F6. One end of each of branch tanks A1, B2, C3, D4, E5, and F6 is connected to the target tank via bolts 12, nuts, and washers. Figure 4 The six vertical connecting walls 8a on the hexagonal G selection pool 8 shown are connected to form a shape as follows: Figure 1-3 The six-way water tank shown.

[0036] Each branch tank has an electric shock terminal 7a at its end, where zebrafish receive electrical pain stimulation. The electric shock terminal 7a is equipped with two electrode plates 11 for providing electrical stimulation to the zebrafish. The electric shock terminal 7a is powered by the two electrode plates 11. The inlet of the electric shock terminal 7a has an infrared beam triggering area for detecting whether a zebrafish has passed by. The electrode plates 11 are connected to an electric shock module for providing voltage to the electrode plates 11. The length × width × height of the electrode plates is 3 × 4 × 0.1 cm.

[0037] The infrared beam triggering area includes an infrared beam sensor 9. Two windows, each 1cm x 2cm in length and width, are positioned face-to-face on both sides of the entrance to the electric shock end 7a. Transparent glass 10s are fixedly connected to each window with adhesive. The outer surface of one transparent glass 10 houses the transmitter of the infrared beam sensor 9, and the outer surface of the other transparent glass 10 houses the receiver. The transmitter of the infrared beam sensor 9 is connected to the power auxiliary module, and the receiver is connected to the main controller. The infrared beam sensor 9 is used to detect whether a zebrafish is passing by. Specifically, the transmitter of the infrared beam sensor 9 emits infrared light, which can pass through the transparent glass 10, and the receiver receives the infrared light in real time. The input to the infrared beam sensor 9 is a regulated power supply (+5V) provided by the power auxiliary module, which continuously emits infrared light signals while simultaneously detecting whether a zebrafish is passing through the channel. Its function is to block the infrared light when a zebrafish passes through the triggering area, thus completing the behavior detection. The output of the infrared beam sensor 9 is to transmit the detected infrared blocking signal (digital pulse) to the main controller as the determination condition for electric shock triggering. That is, the main controller provides electric shock voltage to the electrode plate 11 through operational amplification and bias circuit.

[0038] like Figure 2 A tracking and recording device 14 is installed above the six-way water tank. The tracking and recording device 14 is connected to the six-way water tank via a tripod-shaped mounting bracket 15. The tracking and recording device 14 is used to collect images and videos of zebrafish in each branch of the six-way water tank during the testing process, providing data support for subsequent analysis of the zebrafish's pain response. The process of analyzing the zebrafish's pain response is not within the scope of this technology; this technology only protects a device that can provide electrical stimulation to zebrafish.

[0039] like Figure 1 Branch water tanks 1 (A), 2 (B), 3 (C), 4 (D), 5 (E), and 6 (F) have the same structure, each including a channel section 7b and an electric shock end 7a. The channel section 7b and the electric shock end 7a are interconnected. The end of the channel section 7b is the entrance to the electric shock end 7a. The bottom of the electric shock end 7a is lower than the bottom of the channel section 7b, that is, the electric shock end 7a is 3cm deeper than the channel section 7b.

[0040] Each branch water tank has a slot for placing a partition plate 13 5mm away from the G selection tank 8. The partition plate 13 separates the branch water tank from the G selection tank 8. After the partition plate 13 is placed in the slot, it makes tight contact with the sealing gasket inside the slot, thus disconnecting the branch water tank from the G selection tank 8. Liquid in the G selection tank 8 will not flow into the disconnected branch water tank.

[0041] The six-way water tank is made of translucent but opaque acrylic material.

[0042] like Figure 5 As shown, the electric shock module includes a transformer module, a rectifier and filter circuit, an operational amplifier and bias circuit, a current sampling circuit, a main controller, a power supply auxiliary module, and an OLED display module.

[0043] like Figure 5 As shown, the input terminal of the transformer module is connected to the mains power, the output terminal of the transformer module is connected to the input terminal of the rectifier and filter circuit, the output terminal of the rectifier and filter circuit is connected to the power supply auxiliary module, the power supply auxiliary module is connected to the operational amplifier and bias circuit, the main controller and the OLED display module respectively, and is also used to power the infrared beam sensor 9. The main controller is connected to the input terminal of the operational amplifier and bias circuit, the output terminal of the operational amplifier and bias circuit is connected to both the current sampling circuit and the electrode plate 11, the output terminal of the current sampling circuit is connected to the input terminal of the main controller, and the output terminal of the main controller is connected to the OLED display module. The number of operational amplifier and bias circuits, current sampling circuits, and infrared beam sensors 9 is the same as the number of branch water tanks. Each branch water tank has two electrode plates on its electrode terminal 7a (one electrode plate is connected to the operational amplifier and bias circuit, and the other electrode plate is connected to the ground wire). The main controller is connected to multiple operational amplifier and bias circuits respectively (for simplicity...). Figure 5 Only one operational amplifier and bias circuit, current sampling circuit, and infrared through-beam sensor 9 are shown. In reality, there are six operational amplifier and bias circuits, current sampling circuits, and infrared through-beam sensors 9. Each operational amplifier and bias circuit is connected to the electrode plate on the corresponding electric shock terminal 7a. The current sampling circuit collects the signal in the corresponding circuit and feeds it back to the main controller. The power supply auxiliary module is connected to multiple operational amplifier and bias circuits.

[0044] The transformer module has the following functions: input: AC mains power (220V); function: to achieve isolation through a coupler, and then to convert the high-voltage AC to low-voltage AC suitable for the circuit; output: low-voltage (15V) AC power, which is sent to the rectifier and filter circuit.

[0045] The rectifier and filter circuit (including diodes D1-D4 and capacitors C1-C4) has the following functions: input: low voltage (15V AC) from the transformer output; function: converting AC to pulsating DC through full-bridge rectification, and then smoothing it into DC voltage by filter capacitors C1-C4; output: DC voltage (15V DC), which is sent to the power supply auxiliary module.

[0046] The rectifier and filter circuit consists of full-bridge rectifiers D1-D4 and filter capacitors C1-C4. The specific circuit connection is as follows: Figure 6As shown, the AC power input AC~ terminal is connected to the two AC input terminals of the full-bridge rectifier. After full-wave rectification by D1–D4, a pulsating DC voltage is obtained at the output terminal. The positive terminal of the rectified product serves as the DC output terminal LM2596in+, and the negative terminal serves as the DC output terminal LM2596in−, providing input voltage for the subsequent DC / DC conversion circuit.

[0047] Filter capacitors C1-C4 are connected in parallel at the rectifier output to store energy and reduce output voltage ripple. C1-C4 are preferably electrolytic capacitors with a withstand voltage of not less than 1.5 times the effective value of the AC input voltage. Using them in parallel can effectively reduce the equivalent series resistance and improve the ripple current withstand capability.

[0048] Therefore, the rectifier and filter circuit in this embodiment can stably convert the AC input voltage into DC voltage and significantly reduce ripple, providing a stable input power supply for the subsequent LM2596 step-down circuit (i.e., the LM2596 voltage regulator circuit) and ensuring the reliability of the electric shock module.

[0049] Power Auxiliary Module (LM2596, DC / DC Converter): The power auxiliary module uses the existing LM2596 step-down circuit. Power Auxiliary Module Input: DC voltage (15V DC) provided by the rectifier and filter circuit; Function of the Power Auxiliary Module: Further regulates and steps down the input DC voltage to generate the different power rails required by the circuit; Power Auxiliary Module Output: First auxiliary power supply (serving the analog circuit (op-amp), regulated to +12V, -12V): Provides ±12V to the operational amplifier circuit, specifically connected to V+ (pin 7) and V- (pin 4) of the U2 operational amplifier, and also provides +12V power to the positive rail VCC of the power output stage; also provides +12V to the bias voltage circuit; Second auxiliary power supply (serving the digital circuit (MCU + peripherals), regulated to +5V): Provides +5V power to the main controller, OLED display module, and infrared photoelectric sensor 9.

[0050] The operational amplifier and bias circuit includes a bias voltage circuit and an operational amplifier circuit connected in sequence. The bias voltage circuit includes amplifier OPA197 (U1) and resistors R1-R4, as shown in the specific circuit connection diagram. Figure 6As shown. The input to the bias voltage circuit is the DAC output voltage of the MCU (0.3-2.8V). The function of the bias voltage circuit is to differentially amplify the DAC voltage and add a reference bias Voff to achieve a control voltage output in the range of 0-9.9V. The output of the bias voltage circuit is the drive voltage VDRV, which serves as the input to the operational amplifier circuit. Voff is generated internally by the bias circuit, using two resistors to divide the +5V regulated power supply to obtain a small voltage of 0.3V. In general, Voff = a very small fixed voltage (approximately 0.3V), generated by the bias voltage circuit itself. The function of the bias voltage circuit is to compensate for the zero-point offset of the DAC, ensuring the final output covers the full range of 0-9.9V. The bias voltage circuit mainly consists of the operational amplifier OPA197, input resistors R1 and R3, feedback resistors R2 and R4, and the bias power supply input terminal VOFF. R1 and R3 are connected to the microcontroller's DAC output VDAC and bias power supply VOFF, respectively. Their other ends are connected to the inverting input -IN of operational amplifier OPA197. The non-inverting input +IN of operational amplifier OPA197 is grounded and connected to ground via resistor R4, forming the lower arm of the differential amplifier circuit. The output DRV of operational amplifier OPA197 forms a negative feedback loop with the inverting input via resistor R2, simultaneously outputting to the subsequent current sampling circuit and the electrode plate. The power supply pins V+ and V- of operational amplifier OPA197 are connected to +12V power and ground, respectively.

[0051] In this embodiment, the resistor values ​​are chosen as R1=R3=10log and R2=R4=39.6log, forming a typical differential amplifier with a gain of approximately:

[0052] ;

[0053] Therefore, the output voltage of this circuit ( ) and input voltage VDAC ( ) and bias voltage VOFF ( The relationship between ) can be represented as:

[0054] .

[0055] In terms of hardware connections, the microcontroller's DAC output port is connected to the VDAC input of the bias voltage circuit. The bias power supply VOFF is provided by the bias voltage circuit and connected to its reference input. The output VDRV of the bias voltage circuit is connected to the operational amplifier circuit, and the output of the operational amplifier circuit is connected to electrode plate 11. The operational amplifier circuit simultaneously monitors the current magnitude in real time through a current sampling circuit and feeds it back to the main controller. In this way, while ensuring the voltage output range and stability, the problem of being unable to output close to 0V due to the lower limit of the DAC output can be avoided, ensuring the continuous adjustability of the electrical stimulation intensity.

[0056] Operational amplifier circuit (HTCOP07, transistor power stage): Input: VDRV signal from the bias voltage; Function: Amplify VDRV to drive output stages Q1 and Q2, providing sufficient current drive capability for the load electrode plates; Output: Electrode plate drive voltage (0-9.9V). The operational amplifier circuit includes amplifier HTCOP07 (U2), resistors R5-R12, and transistors Q1-Q2. Specific circuit connections are as follows... Figure 6 As shown. The operational amplifier and bias circuit is used to differentially amplify and power amplify the received voltage, and output a drive voltage to one electrode plate 11. The other electrode plate 11 is connected to the ground wire, forming a circuit through the liquid in the electric shock terminal 7a, thereby realizing electric shock to the zebrafish in the liquid. The operational amplifier circuit is used to effectively drive the subsequent electrode plate 11 to realize the electrical stimulation function.

[0057] The operational amplifier circuit in the operational amplifier and bias circuitry is used to precisely amplify the drive voltage VDRV output from the bias voltage circuit and drive the electrode load, achieving a low-impedance adjustable output of 0~9.9V. This circuit mainly consists of operational amplifier U2, complementary power transistors Q1 and Q2, a base current-limiting resistor, an emitter current-sharing resistor, and a sampling resistor. A compensation capacitor C5 is connected between the output terminal and the inverting input terminal of operational amplifier U2 to ensure the stability of closed-loop operation.

[0058] During circuit operation, VDRV serves as the input to operational amplifier U2, and its output VOUT is connected to the inverting input via feedback. The op-amp automatically adjusts the conduction levels of Q1 and Q2 to ensure that VOUT ≈ VDRV. This allows for continuous adjustment of VOUT (Vout) from 0 to 9.9V within the DAC output range of 0.3 to 2.8V. The emitter resistor improves crossover distortion, while the sampling resistor is used for current monitoring and protection.

[0059] The operational amplifier circuit's input VDRV receives the voltage from the bias voltage circuit, and its output VOUT (Vout) is connected to the electrode plate via a current sampling resistor and fed back to the main controller. Used in conjunction with the bias voltage circuit and the current sampling circuit, it can achieve the high-precision, low-impedance, and stable voltage drive required for electrical stimulation.

[0060] The current sampling circuit includes the current detection chip INA282, and the specific circuit connection method is as follows: Figure 6 As shown. The input of the current sampling circuit is the current in the circuit of electrode 11, which is converted into a voltage difference through the shunt resistor R10. The function of the current sampling circuit is to precisely amplify the shunt voltage using the INA282 to obtain the load current signal. The output of the current sampling circuit is the feedback voltage Vadc, which is sent to the main controller ADC for closed-loop control and safety protection. The current sampling circuit is used to collect the current passing through electrode 11 and feed it back to the main controller.

[0061] The current sampling circuit mainly consists of the current detection chip INA282 and the shunt resistor R10. The shunt resistor R10 is connected in series between the load RL (electrode 11) and the output terminal VOUT (Vout), and is used to convert the current flowing through the load into a small voltage signal. This voltage signal (the voltage across R10 measured by the INA282 voltmeter) is detected and amplified by the differential input terminals +IN and -IN of the INA282, and the corresponding voltage is output on the OUT pin.

[0062] The REF1 and REF2 pins of the INA282 are grounded, making the output reference level 0V, and unidirectional current sampling is used. The chip's power supply terminal V+ is connected to a +12V power supply, and a decoupling capacitor is connected in parallel between V+ and ground to enhance power supply stability. An RC filter network is set at the output terminal before the data is sent to the microcontroller's ADC for acquisition.

[0063] This current sampling circuit can reflect changes in load current in real time, providing a closed-loop feedback signal for the system. Combined with the aforementioned bias circuit and operational amplifier circuit, it can realize the functions of output current detection, control, and protection.

[0064] The main controller includes an STM32CBT6 microcontroller. Its inputs include the feedback voltage Vadc from the current sampling circuit and the detection signal from the infrared photoelectric sensor 9. The main controller's functions include running control algorithms to implement logic such as voltage regulation, timing control, and accidental touch prevention. Its outputs include the DAC output voltage, which drives the bias circuit, and control signals to the OLED display module. The STM32F103CBT6 microcontroller is used as the core controller. The microcontroller communicates with the OLED display module via an I2C bus to display electric shock parameters in real time. It also interacts with a host computer or external control terminal via a serial port to perform parameter setting and data recording functions.

[0065] The OLED display module receives serial communication signals from the main controller (STM32CBT6 microcontroller). Its function is to display the parameters calculated and set by the main controller (voltage value, duration, interval, number of times, etc.) in real time, allowing the operator to observe them intuitively. The OLED display module outputs human-machine interface information (presented visually) and no longer outputs electrical signals to other circuit modules.

[0066] like Figure 7 The software flowchart of the electric shock module shown is provided below. The computer program inside the main controller (STM32CBT6 microcontroller) uses existing technology and is not within the scope of this technical protection. A brief introduction follows. After power-on, the electric shock module performs initialization: It completes hardware configuration such as the microcontroller clock, I / O ports, and interrupts; it initializes parameters such as the DAC output channel, reference voltage, and initial value; it configures the timer's timing cycle; and it initializes the infrared beam sensor 9, ensuring continuous infrared emission. The receiving circuit prepares to collect and analyze the signal. After completion, the main program enters the main loop, monitoring the infrared received signal in real time to determine if there is a zebrafish-triggered infrared blockage. If a suspected zebrafish-triggered infrared signal is detected, the system delays for 10ms for rechecking: if the blockage signal disappears during the recheck, it is determined to be a false trigger, and the system returns to the main loop to continue monitoring; if the signal persists, it is determined to be a valid trigger, the timer is started, and the DAC outputs the target voltage. Simultaneously, the output current is collected in real time through the current sampling circuit, and the sampled value is compared with the target value. A feedback calibration algorithm is used to dynamically adjust the DAC parameters to ensure accurate and stable output. When the DAC outputs the target voltage, the system continuously checks if the timer overflows: if it doesn't overflow, the output and feedback calibration are maintained; if it overflows, the DAC output is controlled to 0V, the output is turned off, and the timer count and trigger flag are reset. The main program then returns to the main loop and continues to monitor the infrared trigger signal in real time, achieving cyclical control of zebrafish trigger detection and DAC output. It should be noted that the duration, intensity, and interval between trigger detection shocks can be flexibly changed by modifying program parameters to adapt to the needs of zebra behavior research in different scenarios.

[0067] In summary, the electric shock module of this technology can not only provide quantitative and adjustable pain stimulation to zebrafish, but also collect feedback data in real time and perform closed-loop control to ensure the stability and repeatability of the output stimulation, thus providing a reliable experimental platform for zebrafish behavioral research and pain response testing.

[0068] In this embodiment, during actual use, a suitable number of branch tanks are selected according to actual needs. Taking three branch tanks as an example, that is, three need to be used (e.g., branch tank 1, branch tank 2, and branch tank 3). The partitions 13 on the remaining three unused branch tanks (e.g., branch tank 4, branch tank 5, and branch tank 6) are closed, i.e., the partitions 13 are inserted into the slots. Liquid (such as conductive water, which mainly refers to an electrolyte solution containing freely moving ions) is placed in the G selection tank 8. The liquid in the G selection tank 8 flows to branch tanks 1, 2, and 3 respectively. Three zebrafish are placed in the G selection tank 8 (depending on actual needs, the three zebrafish can be fed different varieties of Chinese herbal medicines to be selected before being placed in the G selection tank, or one zebrafish can be left unfed to serve as a control). With manual assistance, the zebrafish swim into the channel sections 7b of branch tanks 1, 2, and 3 respectively. One zebrafish is fed by an infrared beam sensor 9 which sends a signal to the main controller in real time. When the main controller determines that a zebrafish has passed by based on the signal from a certain infrared beam sensor 9 (i.e., the zebrafish swims from channel segment 7b to the electric shock end 7a), the main controller sends a signal to the bias voltage circuit in the corresponding operational amplifier and bias circuit. The operational amplifier circuit then provides the voltage required for electric shock to the corresponding electrode 11. After a preset time, the electric shock is stopped. The tracking and recording device 14 records the pain response status of three zebrafish after each of them has been shocked once and compares the results, thereby achieving rapid breeding based on drug efficacy.

[0069] The scope of protection of this technology includes, but is not limited to, the above embodiments. The scope of protection of this technology is defined by the claims. Any substitutions, modifications, or improvements to this technology that are easily conceived by those skilled in the art shall fall within the scope of protection of this technology.

Claims

1. A zebrafish pain response testing device for the selection and breeding of traditional Chinese medicine, characterized in that, It includes multiple branch water tanks, which are connected by a G selection pool (8) to form a six-way water tank. All the branch water tanks are connected to the G selection pool (8). Each branch tank has an electric shock end (7a) at its end for zebrafish to receive electric pain stimulation. The electric shock end (7a) is provided with an electrode plate (11) for providing electric stimulation to the zebrafish. The entrance of the electric shock end (7a) is provided with an infrared beam triggering area for detecting whether a zebrafish has passed by. The electrode plate (11) is connected to an electric shock module for providing voltage to the electrode plate (11).

2. The zebrafish pain response testing device for the breeding of traditional Chinese medicines according to claim 1, characterized in that, There are six branch water tanks, which are respectively labeled as branch water tank A (1), branch water tank B (2), branch water tank C (3), branch water tank D (4), branch water tank E (5) and branch water tank F (6). Branch water tank A (1), branch water tank B (2), branch water tank C (3), branch water tank D (4), branch water tank E (5) and branch water tank F (6) are connected to the regular hexagonal G selection pool (8) by bolts (12), nuts and washers respectively.

3. The zebrafish pain response testing device for the breeding of traditional Chinese medicines according to claim 1, characterized in that, Each branch water tank is provided with a slot for placing the partition (13) 5 mm away from the G selection pool (8).

4. The zebrafish pain response testing device for the breeding of traditional Chinese medicines according to claim 1, characterized in that, Each branch tank includes a channel section (7b) and an electric shock end (7a), which are connected. The end of the channel section (7b) is the inlet of the electric shock end (7a), and the bottom of the electric shock end (7a) is lower than the bottom of the channel section (7b).

5. The zebrafish pain response testing device for the breeding of traditional Chinese medicines according to claim 1, characterized in that, The electric shock module includes a transformer module, a rectifier and filter circuit, an operational amplifier and bias circuit, a main controller, a power supply auxiliary module, and an OLED display module. The input terminal of the transformer module is connected to the mains power, the output terminal of the transformer module is connected to the input terminal of the rectifier and filter circuit, the output terminal of the rectifier and filter circuit is connected to the power supply auxiliary module, the power supply auxiliary module is connected to the operational amplifier and bias circuit, the main controller and the OLED display module respectively, the main controller is connected to the input terminal of the operational amplifier and bias circuit, the output terminal of the operational amplifier and bias circuit is connected to the current sampling circuit and the electrode plate (11) at the same time, the output terminal of the current sampling circuit is connected to the input terminal of the main controller, and the output terminal of the main controller is connected to the OLED display module.

6. The zebrafish pain response testing device for the breeding of traditional Chinese medicines according to claim 5, characterized in that, The infrared beam triggering area includes an infrared beam sensor (9). On the two side walls of the electric shock end (7a) entrance, there are windows facing each other. Transparent glass (10) is fixedly connected to each of the two windows. The outer side of one transparent glass (10) is provided with the transmitting end of the infrared beam sensor (9), and the outer side of the other transparent glass (10) is provided with the receiving end of the infrared beam sensor (9). The transmitting end of the infrared beam sensor (9) is connected to the power auxiliary module, and the receiving end of the infrared beam sensor (9) is connected to the main controller. The infrared beam sensor (9) is used to detect whether a zebrafish has passed by.

7. The zebrafish pain response testing device for the breeding of traditional Chinese medicines according to claim 6, characterized in that, The transformer module is used to receive mains power and convert it into 15V AC voltage. The rectifier and filter circuit is used to convert the 15V AC voltage into 15V DC voltage. The power supply auxiliary module uses an LM2596 step-down circuit to convert the 15V DC voltage into 12V and 5V voltages. The 12V voltage is used to input the operational amplifier and bias circuit, and the 5V voltage is used to input the main controller, OLED display module, and infrared beam sensor (9). The operational amplifier and bias circuit includes a bias voltage circuit and an operational amplifier circuit connected in sequence. The bias voltage circuit includes an amplifier OPA197, and the operational amplifier circuit includes an amplifier HTCOP07. The operational amplifier and bias circuit is used to differentially amplify the received voltage and... The power is amplified and the driving voltage is output to one electrode plate (11). The other electrode plate (11) is connected to the ground wire. The two electrode plates (11) are respectively connected to the inner wall of the electric shock end (7a). The current sampling circuit includes a current detection chip INA282. The current sampling circuit is used to collect the current passing through the electrode plate (11) and feed it back to the main controller. The main controller includes a microcontroller STM32CBT6. The main controller is used to receive the signal sent by the infrared beam sensor (9) and analyze and judge it. It is also used to send signals to the operational amplifier and bias circuit and the OLED display module respectively. The OLED display module is used to display the information collected by the current sampling circuit and the working time of the electrode plate (11) according to the control instructions of the main controller.

8. The zebrafish pain response testing device for the breeding of traditional Chinese medicines according to claim 1, characterized in that, A tracking camera (14) is installed above the six-way water tank.

9. The zebrafish pain response testing device for the breeding of traditional Chinese medicines according to claim 8, characterized in that, The tracking and recording device (14) is connected to the six-way water tank via a tripod-shaped mounting bracket (15).

10. The zebrafish pain response testing device for the breeding of traditional Chinese medicines according to claim 1, characterized in that, The six-way water tank is made of translucent but opaque acrylic material.

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