Full-automatic electrochemical luminescence analyzer based on automatic in and out of chip

Abstract

The utility model discloses a full-automatic electrochemical luminescence analyzer based on automatic chip in and out, which comprises a shell, an automatic chip in and out unit, an electrochemical luminescence detection chip, a circuit board, a camera component, a connecting device, a lithium battery pack and a microprocessor, and the lithium battery pack is connected with the circuit board; the shell comprises an upper cover, a cavity, a fixing device and a base, the upper cover and the base are connected with the cavity, the fixing device is arranged in the cavity, the lithium battery pack and the camera assembly are arranged on the fixing device, a button, an adjustable potentiometer and an excitation switch are arranged on the surface of the shell, and the button, the adjustable potentiometer and the excitation switch are connected with the circuit board; the chip automatic in-out unit is used for transmitting an electrochemical luminescence detection chip to a detection area, the electrochemical luminescence excitation unit is used for exciting a sample to be detected to generate an electrochemical luminescence signal, the camera assembly is used for capturing the electrochemical luminescence signal and transmitting the electrochemical luminescence signal to the microprocessor, and the microprocessor is used for processing the signal transmitted by the camera assembly. And calculating an electrochemical luminescence intensity value.

Description

Technical Field

[0001] The utility model relates to the technical field of electrochemiluminescence analyzers, and in particular to a full-automatic electrochemiluminescence analyzer based on automatic chip entry and exit. Background Art

[0002] Electrochemiluminescence (ECL) detection is an analytical technique that combines electrochemistry and chemiluminescence. It generates luminescent excited-state species by inducing an electrochemical reaction on an electrode surface. When these excited-state species return to their ground state, they release photons, generating luminescence. By measuring the intensity of this luminescence, specific components in a sample can be quantitatively or qualitatively analyzed. With its low background signal, high sensitivity, excellent selectivity, and wide linear range, ECL detection is particularly suitable for detecting markers such as proteins, nucleic acids, and cells.

[0003] Currently, fully automated ECL analyzers are typically bulky, difficult to port, and expensive. Small ECL analyzers, on the other hand, have a low degree of automation, and their chip loading requires manual operation, which significantly impacts detection errors. Furthermore, some ECL analyzers on the market have relatively independent electrochemical excitation units, control units, and data processing units, resulting in low levels of integration and automation. The detection process and operating steps are cumbersome, requiring high operational requirements, and therefore lack the potential for application in homes and community medical institutions. Summary of the Invention

[0004] The purpose of the utility model is to overcome the shortcomings and deficiencies of the prior art and provide a fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit, which is automatic and portable, simple to assemble, easy to operate, and has low production cost.

[0005] In order to achieve the above purpose, the technical solutions adopted by the present invention are as follows:

[0006] A fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit, comprising a housing, an automatic chip entry and exit unit, an electrochemiluminescence detection chip, a circuit board, a camera assembly, a connecting device, a lithium battery pack and a microprocessor, wherein the lithium battery pack is connected to the circuit board;

[0007] The housing includes an upper cover, a cavity, a fixing device and a base. The upper cover and the base are respectively connected to the cavity. The fixing device is arranged in the cavity. The lithium battery pack and the camera assembly are arranged on the fixing device. The surface of the housing is provided with a button, an adjustable potentiometer and an excitation switch.

[0008] The chip automatic entry and exit unit is fixed on the base, and the chip automatic entry and exit unit includes a micro stepping motor and a chip placement tray. The micro stepping motor is arranged below the chip placement tray, and the electrochemiluminescence detection chip is arranged on the chip placement tray. The electrochemiluminescence detection chip is provided with an electrochemiluminescence reaction cell for the object to be tested;

[0009] The circuit board is fixed on the base. The circuit board integrates a power module, a drive circuit, a main control chip and an electrochemical luminescence excitation unit. The power module is connected to the micro stepping motor and the main control chip respectively. The drive circuit is connected to the micro stepping motor and the main control chip respectively. The main control chip is connected to the button. The electrochemical luminescence excitation unit is connected to the adjustable potentiometer and the excitation switch respectively.

[0010] The camera assembly includes a UVC camera and an industrial lens. The UVC camera is connected to a microprocessor. The industrial lens faces the electrochemiluminescence reaction cell of the object to be tested. The connecting device is fixed to the chip placement plate. The connecting device includes a connector and a spring-type contact. The connector is coupled to the electrochemiluminescence detection chip. The spring-type contact is respectively connected to the electrode of the electrochemiluminescence reaction cell of the object to be tested and the output end of the electrochemiluminescence excitation unit.

[0011] The chip automatic entry and exit unit is used to automatically transfer the electrochemiluminescence detection chip to the detection area below the fixing device. The electrochemiluminescence excitation unit is used to excite the sample to be tested to generate an electrochemiluminescence signal. The camera component is used to capture the electrochemiluminescence signal and convert it into an electrical signal and transmit it to the microprocessor. The microprocessor is used to process the electrical signal transmitted by the camera component, calculate the electrochemiluminescence intensity value and display the calculation result.

[0012] Furthermore, the shell and the chip placement tray are both made of 1.75mm black polylactic acid PLA material through 3D printing technology, and the filling density of the polylactic acid PLA material is 80%.

[0013] Furthermore, the fixing device includes a fixing frame and a light shielding plate, the light shielding plate is perpendicular to the fixing frame and is located above the chip automatic entry and exit unit, and the light shielding plate is provided with a through hole facing the electrochemiluminescence reaction cell of the object to be tested.

[0014] Furthermore, the power module provides power for the micro stepper motor and main control chip. The main control chip adopts the STM32 main control chip. The 24V input power is converted from 24V to 7V through a step-down circuit based on the LM2596S-ADJ voltage regulator chip design and 7V regulated output, providing a 7V regulated power supply for the micro stepper motor.

[0015] The power supply of the main control chip achieves a 3.3V regulated output through three-way step-down conversion. Through the voltage stabilization circuit designed based on three voltage stabilization chips LM2596S-ADJ, LM2596S-5.0, and LM2596S-3.3, it realizes 24V to 7V, then from 7V to 5V, and finally from 5V to 3.3V, providing a 3.3V regulated power supply for the main control chip.

[0016] Furthermore, the circuit board also integrates a decoupling circuit, a crystal oscillator circuit, a reset circuit, a start-up circuit, a debugging interface circuit, and a connection circuit. The micro stepping motor is connected to the driving circuit through the connection circuit, and the decoupling circuit, crystal oscillator circuit, reset circuit, start-up circuit, and debugging interface circuit are respectively connected to the main control chip.

[0017] Furthermore, the decoupling circuit is composed of five 100nF chip capacitors in a 0805 package. The decoupling circuit provides a low-impedance power path for the main control chip and filters out transient current noise generated by the switching of logic gates inside the main control chip.

[0018] The crystal oscillator circuit uses crystal oscillators with frequencies of 8MHz and 32.768kHz. Two identical load capacitors are set at both ends of the crystal oscillator. A load capacitor of 22pF is set at both ends of the 8MHz crystal oscillator, and a load capacitor of 8pF is set at both ends of the 32.768kHz crystal oscillator.

[0019] Furthermore, the reset circuit uses a low-level reset, and a capacitor is connected in parallel with the button for button debounce; when the button is turned off, the capacitor is charged and RST is at a high level. When the button is pressed, the capacitor releases current, and the resistor in series with the capacitor will keep the current on the NRST pin from exceeding 3.3V when the switch SW1 is closed; after the button is restored, the capacitor will be charged again; during the key restart period, the NRST pin remains at a low level.

[0020] Furthermore, the boot circuit is a BOOT boot circuit, and the main flash memory boot mode is selected. A 2×3P pin header is used, and pins 1 and 2 of the pin header are connected to a 3.3V voltage, pins 5 and 6 are connected to the ground line, and the boot configuration pins BOOT0 and BOOT1 are connected to pins 3 and 4 of the pin header respectively;

[0021] Connect pins 3 and 5, and pins 4 and 6 so that the boot configuration pins BOOT0 and BOOT1 are both equal to 0 to implement the main flash memory boot mode. In addition, connect 10kΩ pull-down resistors between the boot configuration pins BOOT0 and 3, and between the boot configuration pins BOOT1 and 4 to limit the current and protect the main control chip.

[0022] Furthermore, the debug interface circuit is a SWD debug interface circuit. The debug interface circuit reads and writes the registers and memory of the main control chip through the SWDIO pin and SWCLK pin. The SWDIO pin is used for bidirectional data transmission, and the SWCLK pin is used to maintain clock synchronization; the connection circuit uses a 4-pin wire-to-wire connector with a built-in spacing of 2mm.

[0023] Furthermore, the driving circuit is an A4988 driving circuit, which uses two 1×8P connectors; the electrochemical luminescence excitation unit includes an LM2596S-ADJ voltage regulator chip, which realizes a 1.2V-24V wide range adjustable regulated power supply output through the LM2596S-ADJ voltage regulator chip and an adjustable potentiometer.

[0024] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0025] 1. The utility model couples the connection device and the chip automatic entry and exit unit together, which enhances the stability of movement and electrical connection and the adaptability to different electrochemiluminescence detection chips. It is suitable for a variety of different types of electrochemiluminescence detection chips and has the advantages of long service life, high flexibility and high integration.

[0026] 2. The utility model highly integrates the electrochemiluminescence excitation unit and the main control chip on the circuit board to form the integrated control system circuit board of the utility model; the circuit board is placed inside the analyzer, avoiding the inconvenience caused by placing the electrochemiluminescence excitation unit outside the analyzer.

[0027] 3. The utility model adopts spring-type contacts for electrical connection to replace the traditional manual alligator clip connection, which improves the stability and convenience of the electrical connection and reduces the error caused by manual placement of the electrochemical luminescence detection chip; the shell draws on the design ideas and methods of the mortise and tenon structure, and is simple and convenient to assemble.

[0028] 4. The analyzer of this utility model is more miniaturized, highly integrated, automated, and portable, and is easy to operate and use. It uses PLA material through 3D printing, which has low material cost and is easy to mass produce and assemble. It only takes about 3 minutes from electrochemiluminescence signal acquisition to analysis, which is convenient to operate and significantly reduces analysis time. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Schematic diagram of the cross-sectional structure of the fully automatic electrochemiluminescence analyzer.

[0030] Figure 2 Schematic diagram of the base assembly structure.

[0031] Figure 3 This is the schematic diagram of the power module circuit.

[0032] Figure 4 This is the schematic diagram of the decoupling circuit.

[0033] Figure 5 This is the schematic diagram of the crystal oscillator circuit.

[0034] Figure 6 This is the reset circuit schematic.

[0035] Figure 7 This is the schematic diagram of the BOOT startup circuit.

[0036] Figure 8 This is the schematic diagram of the SWD debug interface circuit.

[0037] Figure 9 This is the driving circuit schematic.

[0038] Figure 10 The following is the connection circuit schematic diagram.

[0039] Figure 11 This is the circuit schematic of the electrochemiluminescence excitation unit.

[0040] Figure 12 A closed bipolar electrode electrochemiluminescence detection chip uses the Ru(bpy)32+ system to detect tripropylamine (TPA).

[0041] Figure 13 It is a closed bipolar electrode electrochemiluminescence detection chip used to detect luteinizing hormone (LH).

[0042] Description of Figure Numbers:

[0043] Housing 1; upper cover 11; cavity 12; fixture 13; light baffle 131; base 14; button 15; adjustable potentiometer 16; voltage digital display 17; excitation switch 18; automatic chip entry and exit unit 2; micro-stepping motor 21; chip placement tray 22; electrochemiluminescence detection chip 3; electrochemiluminescence reaction cell for test article 31; circuit board 4; drive circuit 41; main control chip 42; electrochemiluminescence excitation unit 43; power input interface 44; camera assembly 5; UVC camera 51; industrial lens 52; connecting device 6; connecting piece 61; spring-loaded copper contact 62; lithium battery pack 7; microprocessor 8; USB interface 9. DETAILED DESCRIPTION

[0044] The following is a further description of the fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit according to the present invention in conjunction with the accompanying drawings and specific embodiments.

[0045] See also Figure 1 and Figure 2The utility model discloses a fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit. The fully automatic electrochemiluminescence analyzer includes a cylindrical shell 1, an automatic chip entry and exit unit 2, an electrochemiluminescence detection chip 3, a circuit board 4, a camera assembly 5, a connecting device 6, a 24V lithium battery pack 7 and a microprocessor 8. The microprocessor 8 can be a smartphone or an Orange Pie processor. The 24V lithium battery pack 7 is connected to the circuit board 4.

[0046] The housing 1 includes a cover 11, a cavity 12, a fixture 13, and a base 14. The cover 11 and base 14 are respectively connected to the cavity 12. The fixture 13 is disposed within the cavity 12, and the 24V lithium battery pack 7 and camera assembly 5 are mounted on the fixture 13. The surface of the housing 1 is provided with a button 15, an adjustable potentiometer 16, a voltage digital display 17, and an excitation switch 18.

[0047] The chip automatic entry and exit unit 2 is fixed on the base 14. The chip automatic entry and exit unit 2 includes a micro stepping motor 21 and a chip placement tray 22. The micro stepping motor 21 is arranged under the chip placement tray 22. The electrochemiluminescence detection chip 3 is arranged on the chip placement tray 22. The electrochemiluminescence detection chip 3 is provided with an electrochemiluminescence reaction pool 31 of the item to be tested.

[0048] like Figure 1 and Figure 2 As shown, the circuit board 4 is fixed to the base 14. The circuit board 4 integrates a power module, a decoupling circuit, a crystal oscillator circuit, a reset circuit, a startup circuit, a debugging interface circuit, a drive circuit 41, a main control chip 42, a connection circuit, and an electrochemical excitation unit 43. The micro-stepping motor 21 is connected to the drive circuit 41 via the connection circuit, while the decoupling circuit, crystal oscillator circuit, reset circuit, startup circuit, debugging interface circuit, and drive circuit 41 are each connected to the main control chip 42. The circuit board 4 also has a power input interface 44, to which the 24V lithium battery pack 7 is connected via wires.

[0049] like Figure 2 As shown, the camera assembly 5 includes a UVC camera 51 and an industrial lens 52. The UVC camera 51 is connected to the microprocessor 8 via a USB data cable via a USB port 9. The industrial lens 52 is facing the electrochemiluminescence reaction cell 31 of the object to be tested. The connecting device 6 is fixed to the chip placement tray 22. The connecting device 6 includes a connector 61 and a spring-loaded copper contact 62. The connector 61 is coupled to the electrochemiluminescence detection chip 3. The spring-loaded copper contact 62 is respectively connected to the electrodes of the electrochemiluminescence reaction cell 31 of the object to be tested and the output end of the electrochemical excitation unit 43.

[0050] A button 15 is connected to a main control chip 42 on a circuit board 4. This main control chip 42 drives a drive circuit 41 connected to a micro-stepping motor 21, controlling the chip to automatically enter and exit the electrochemiluminescence detection chip 3 on the unit 2. An adjustable potentiometer 16 is connected to an electrochemical excitation unit 43 on the circuit board 4. The adjustable potentiometer 16 is used to adjust the electrochemiluminescence excitation voltage, which is displayed in real time on a voltage digital display 17 connected to the electrochemical excitation unit 43. An excitation switch 18 is connected to the electrochemical excitation unit 43 on the circuit board 4 and controls the generation and disconnection of the electrochemiluminescence voltage.

[0051] The chip automatic entry and exit unit 2 is used to automatically transfer the electrochemiluminescence detection chip 3 to the detection area below the fixing device 13, the electrochemical excitation unit 43 is used to excite the sample to be tested to generate an electrochemiluminescence signal, the camera component 5 is used to capture the electrochemiluminescence signal and convert it into an electrical signal and transmit it to the microprocessor 8, the microprocessor 8 is used to process the electrical signal transmitted by the camera component 5, calculate the electrochemiluminescence intensity value and display the calculation result.

[0052] like Figure 1 As shown, the fixing device 13 includes a fixing frame and a light shield 131. The light shield 131 is perpendicular to the fixing frame and is located above the chip automatic entry and exit unit 2. The light shield 131 is provided with a through hole facing the electrochemiluminescence reaction pool 31 of the object to be tested. The industrial lens 52 enters the through hole to capture the electrochemiluminescence signal in the electrochemiluminescence reaction pool 31 of the object to be tested.

[0053] like Figure 3 As shown, the power module provides power for the micro-stepping motor 21 and the main control chip 42. The main control chip 42 uses an STM32 main control chip. The 24V input power is converted from 24V to 7V through a step-down circuit based on the LM2596S-ADJ voltage regulator chip design and a 7V regulated output, providing a 7V regulated power supply for the micro-stepping motor 21.

[0054] To prevent excessive power consumption and heat generation when the 24V-to-3.3V step-down chip is directly converted, the power supply for the main control chip 42 is achieved through a three-way step-down conversion to a 3.3V regulated output. This voltage-stabilizing circuit, designed based on the LM2596S-ADJ, LM2596S-5.0, and LM2596S-3.3 voltage regulator chips, converts 24V to 7V, then 7V to 5V, and finally 5V to 3.3V, providing a 3.3V regulated power supply for the main control chip 42.

[0055] like Figure 4As shown, the decoupling circuit is composed of five 100nF chip capacitors in a 0805 package. The decoupling circuit provides a power supply path with a lower impedance for the main control chip 42 and filters out transient current noise generated by the switching of internal logic gates of the main control chip 42 to provide a stable power supply while reducing the impact on other components.

[0056] like Figure 5 As shown in the figure, the crystal oscillator circuit uses crystal oscillators with frequencies of 8MHz and 32.768kHz. Two identical load capacitors are set at both ends of the crystal oscillator. A load capacitor of 22pF is set at both ends of the 8MHz crystal oscillator, and a load capacitor of 8pF is set at both ends of the 32.768kHz crystal oscillator. The resonance condition is met and positive feedback is formed to ensure the stability of the oscillation frequency.

[0057] like Figure 6 As shown in the figure, the reset circuit uses a low-level reset, and a capacitor is connected in parallel with the key to debounce the key. When the key KEY1 is turned off, capacitor C19 is charged and the NRST pin is at a high level. When the key KEY1 is pressed, capacitor C19 releases current. Resistor R5, connected in series with the capacitor, keeps the current on the NRST pin below 3.3V when the switch key KEY1 is turned off. After the key KEY1 is restored, capacitor C19 is charged again. During the hundreds of nanoseconds when the key KEY1 is restarted, the NRST pin remains at a low level, generating a system reset.

[0058] like Figure 7 As shown in the figure, the boot circuit is a BOOT boot circuit, and the main flash memory boot mode is selected. A 2×3P pin header is used, and pins 1 and 2 of the pin header are connected to a 3.3V voltage, pins 5 and 6 are connected to the ground (GND), and the boot configuration pins BOOT0 and BOOT1 are connected to pins 3 and 4 of the pin header respectively.

[0059] Use electronic component jumper caps to connect pins 3 and 5, and pins 4 and 6, so that the boot configuration pins BOOT0 and BOOT1 are both equal to 0, so as to realize the main flash memory boot mode, and connect 10kΩ pull-down resistors between the boot configuration pins BOOT0 and 3, and between the boot configuration pins BOOT1 and 4, respectively, to limit the current and protect the main control chip 42.

[0060] like Figure 8As shown, the debug interface circuit is a SWD debug interface circuit. SWD is a serial single-wire interface used for program debugging. The debug interface circuit reads and writes the registers and memory of the main control chip 42 through the SWDIO pin and SWCLK pin. The SWDIO pin is used for bidirectional data transmission, and the SWCLK pin is used to maintain clock synchronization. The interface can be used to quickly and efficiently perform program debugging, optimization, and troubleshooting.

[0061] like Figure 9 As shown, the driving circuit 41 is an A4988 driving circuit, and the driving circuit 41 uses two 1×8P connectors to facilitate the connection of the A4988 driver.

[0062] like Figure 10 As shown, according to the interface specifications of the micro stepper motor 21, the connection circuit adopts a 4-pin wire-to-wire connector with a built-in spacing of 2mm to ensure that the four signal lines of the micro stepper motor 21 are efficiently and directly connected to the circuit board 4 through the connector.

[0063] like Figure 11 As shown, the electrochemical excitation unit 43 also includes an LM2596S-ADJ voltage regulator chip. Through the LM2596S-ADJ voltage regulator chip and the adjustable potentiometer 16, a wide range of adjustable regulated power supply output of 1.2V-24V is achieved to meet the needs of electrical signal excitation of the fully automatic electrochemiluminescence analyzer.

[0064] Both the housing 1 and the chip tray 22 are made of 1.75mm thick black polylactic acid (PL) material using 3D printing technology, with a fill density of 80%. In this fully automatic electrochemiluminescence analyzer, the housing 1's upper cover 11 and the bulk of the cavity 12 are cylindrical in dimensions of 136mm x 189.5mm. The 24V lithium battery pack 7 charges in 2.5 hours and offers up to four hours of uninterrupted use.

[0065] Application Example 1

[0066] This example uses a fully automatic electrochemiluminescence analyzer to analyze Ru(bpy)3 2+ The system detects tripropylamine (TPA) using a closed bipolar electrode electrochemiluminescence detection chip.

[0067] (1) Prepare 2.5 mM Ru(bpy)3 using purified water 2+ solution, and then 10-fold diluted phosphate buffer solution (PBS) was used to prepare 0.1, 0.5, 1, 2.5 and 5 mM tripropylamine (TPA) solutions, and mixed with 2.5 mM Ru(bpy)3 2+ The solutions were mixed in a 1:1 volume ratio to obtain the test solution.

[0068] (2) Use a pipette to add 40 μL of phosphate buffered saline (PBS) to the sample well of the support channel of the closed bipolar electrode electrochemiluminescence detection chip, and then add 40 μL of the test solution to the sample well of the reporter channel.

[0069] (2) The closed bipolar electrode electrochemiluminescence detection chip enters the analyzer through the chip automatic entry and exit unit.

[0070] (3) Open the video acquisition function in the mobile phone APP of the electrochemiluminescence analyzer, set the white balance to 2800K and the exposure time to 166ms, click the video recording icon, and apply a 7V excitation voltage at the same time.

[0071] (4) After the luminescence ends, click the video recording icon again and the electrochemiluminescence signal video will be saved in the mobile phone album.

[0072] (5) Open the APP video analysis function and convert the electrochemiluminescence video signal into electrochemiluminescence intensity value. The experimental results are as follows Figure 12 shown.

[0073] As can be seen from the figure, using the fully automatic electrochemiluminescence analyzer of the present invention, as the concentration of tripropylamine (TPA) increases, the electrochemiluminescence intensity value also increases accordingly; and there is a good linear relationship between the tripropylamine (TPA) concentration and the electrochemiluminescence intensity value, the linear fitting equation is Y = 3.851X + 0.057, and the correlation coefficient R 2 is 0.9876 (5 repetitions). Therefore, the fully automatic electrochemiluminescence analyzer of the present invention can be applied to Ru(bpy)3 2+ The system detects tripropylamine (TPA).

[0074] Application Example 2

[0075] In this example, a fully automatic electrochemiluminescence analyzer was used for immunoassay. The immunoassay target was luteinizing hormone (LH), and a closed dual-electrode electrochemiluminescence detection chip was used.

[0076] (1) Preparation of sample pad

[0077] 200 μL of sample pad treatment solution (0.025% casein, 0.05% polyvinylpyrrolidone (PVP), 0.25% polysorbate 20 (S19) and 10-fold diluted phosphate buffer solution (PBS)) was added dropwise onto a 1×5 cm glass fiber and baked in a 37°C oven for 1 h for later use.

[0078] (2) Preparation of conjugate pad

[0079] 100 μL of conjugate pad treatment solution (0.025% casein, 0.05% PVP, 0.25% S19, and 10-fold diluted phosphate buffer solution (PBS)) was added dropwise to a 0.5×5 cm glass fiber and baked in a 37°C oven for 1 hour. Next, the prepared T-line and C-line electrochemiluminescent probes were mixed at a volume ratio of 2:1, and the mixture contained 0.05% PVP, 5 mg / mL trehalose, 0.03% proclin-300, 0.025% casein, and 0.1% S9. The mixed probes were then sprayed onto the conjugate pad using a gold sprayer at a spray volume of 6 μL / cm and a speed of 50 mm / s (repeated twice). Finally, the conjugate pad was baked at 37°C for 1 hour and set aside.

[0080] (3) Processing the detection pad

[0081] The C-line capture antibody (5 mg / mL trehalose, 0.1% S9, 5 mg / mL sucrose, and 0.25 mg / mL antibody dissolved in phosphate buffered saline (PBS) at pH 7) and the T-line capture antibody (0.1% S9, 15 mg / mL sucrose, and 0.5 mg / mL antibody dissolved in phosphate buffered saline (PBS) at pH 7) were used. Subsequently, the T-line and C-line capture antibodies were coated onto a nitrocellulose membrane (NC membrane) using a gold sprayer at 50 mm / s and a spray volume of 2 μL / cm (repeated twice). Finally, the NC membrane was baked at 37°C for 2 hours and used.

[0082] (4) Preparation of electrode pads

[0083] First, a screen printing plate is mounted on a screen printing table. A hydrophobic PET substrate is then placed beneath the screen printing plate. Subsequently, a conductive carbon paste is poured onto the screen printing plate. The carbon paste is then squeezed onto the PET substrate using a screen printing scraper. Finally, the screen-printed PET substrate is dried overnight at room temperature to obtain the prepared electrode pad, which is then ready for use.

[0084] (5) Preparation of connection pads

[0085] Cut out 0.5×1 cm absorbent paper to form the connection pad.

[0086] (6) Preparation of absorbent pad

[0087] Cut out 5×0.8 cm pieces of absorbent paper to make the absorbent pad.

[0088] (7) Assembling a dry closed bipolar electrode electrochemiluminescent lateral flow immunoassay test strip, the process is as follows:

[0089] First, stick the NC membrane (detection pad) on the PET base plate. Then, stack the absorption pad near the T line edge, overlapping by about 1mm. Next, stack the binding pad and sample pad in sequence near the C line edge, overlapping by about 1mm. Subsequently, cut the assembled test strips into strips. Next, turn it upside down on the electrode pad so that the C line and T line overlap on the two anodes of the electrode respectively, and stick the connection pad on the closed bipolar electrode cathode and the corresponding positive drive electrode. Finally, assemble the closed bipolar electrode electrochemiluminescence lateral flow immunoassay strip with its housing for luteinizing hormone (LH) immunoassay.

[0090] (8) The concentrations of luteinizing hormone (LH) samples used in the experiment were 0.1, 0.5, 1, 2.5, and 11 mIU / mL.

[0091] The experimental results are as follows Figure 13 As shown in FIG, it can be seen from the test results that: using the fully automatic electrochemiluminescence analyzer of the present invention, as the concentration of luteinizing hormone (LH) increases, the ratio of the electrochemiluminescence intensity value on the chip T line to the electrochemiluminescence intensity value on the C line (i.e., T / C) also increases accordingly; and there is a good linear relationship between the logarithm of the luteinizing hormone (LH) concentration and T / C, the linear fitting equation is Y = 1.961X + 2.662, and the correlation coefficient R 2 The value is 0.9985 (5 repetitions). Therefore, the fully automatic electrochemiluminescence analyzer of the present invention can be applied to the immunoassay of luteinizing hormone (LH), and can also be applied to the immunoassay of biomarkers of other diseases and other physiological activities.

[0092] In summary, the utility model has the following advantages and beneficial effects:

[0093] 1. The utility model couples the connection device and the chip automatic entry and exit unit together, which enhances the stability of movement and electrical connection and the adaptability to different electrochemiluminescence detection chips. It is suitable for a variety of different types of electrochemiluminescence detection chips and has the advantages of long service life, high flexibility and high integration.

[0094] 2. The utility model highly integrates the electrochemiluminescence excitation unit and the main control chip on the circuit board to form the integrated control system circuit board of the utility model; the circuit board is placed inside the analyzer, avoiding the inconvenience caused by placing the electrochemiluminescence excitation unit outside the analyzer.

[0095] 3. The utility model adopts spring-type contacts for electrical connection to replace the traditional manual alligator clip connection, which improves the stability and convenience of the electrical connection and reduces the error caused by manual placement of the electrochemical luminescence detection chip; the shell draws on the design ideas and methods of the mortise and tenon structure, and is simple and convenient to assemble.

[0096] 4. The analyzer of this utility model is more miniaturized, highly integrated, automated, and portable, and is easy to operate and use. It uses PLA material through 3D printing, which has low material cost and is easy to mass produce and assemble. It only takes about 3 minutes from electrochemiluminescence signal acquisition to analysis, which is convenient to operate and significantly reduces analysis time.

[0097] The above description is a detailed description of the preferred feasible embodiment, but the embodiment is not intended to limit the scope of the patent application. All equivalent changes or modifications made under the disclosed technical spirit should fall within the scope of the patent.

Claims

1. A fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit, characterized in that: It includes a housing, an automatic chip entry and exit unit, an electrochemical luminescence detection chip, a circuit board, a camera assembly, a connecting device, a lithium battery pack and a microprocessor, wherein the lithium battery pack is connected to the circuit board; The housing includes an upper cover, a cavity, a fixing device and a base. The upper cover and the base are respectively connected to the cavity. The fixing device is arranged in the cavity. The lithium battery pack and the camera assembly are arranged on the fixing device. The surface of the housing is provided with a button, an adjustable potentiometer and an excitation switch. The chip automatic entry and exit unit is fixed on the base, and the chip automatic entry and exit unit includes a micro stepping motor and a chip placement tray. The micro stepping motor is arranged below the chip placement tray, and the electrochemiluminescence detection chip is arranged on the chip placement tray. The electrochemiluminescence detection chip is provided with an electrochemiluminescence reaction cell for the object to be tested; The circuit board is fixed on the base. The circuit board integrates a power module, a drive circuit, a main control chip and an electrochemical luminescence excitation unit. The power module is connected to the micro stepping motor and the main control chip respectively. The drive circuit is connected to the micro stepping motor and the main control chip respectively. The main control chip is connected to the button. The electrochemical luminescence excitation unit is connected to the adjustable potentiometer and the excitation switch respectively. The camera assembly includes a UVC camera and an industrial lens. The UVC camera is connected to a microprocessor. The industrial lens faces the electrochemiluminescence reaction cell of the object to be tested. The connecting device is fixed to the chip placement plate. The connecting device includes a connector and a spring-type contact. The connector is coupled to the electrochemiluminescence detection chip. The spring-type contact is respectively connected to the electrode of the electrochemiluminescence reaction cell of the object to be tested and the output end of the electrochemiluminescence excitation unit. The chip automatic entry and exit unit is used to automatically transfer the electrochemiluminescence detection chip to the detection area below the fixing device. The electrochemiluminescence excitation unit is used to excite the sample to be tested to generate an electrochemiluminescence signal. The camera component is used to capture the electrochemiluminescence signal and convert it into an electrical signal and transmit it to the microprocessor. The microprocessor is used to process the electrical signal transmitted by the camera component, calculate the electrochemiluminescence intensity value and display the calculation result.

2. A fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit according to claim 1, characterized in that: The shell and chip placement tray are made using 3D printing technology.

3. The fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit according to claim 1, characterized in that: The fixing device includes a fixing frame and a light shielding plate. The light shielding plate is perpendicular to the fixing frame and is located above the chip automatic entry and exit unit. The light shielding plate is provided with a through hole facing the electrochemical luminescence reaction pool of the object to be tested.

4. The fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit according to claim 1, characterized in that: The power module provides power for the micro stepper motor and main control chip. The main control chip adopts the STM32 main control chip. The 24V input power is converted from 24V to 7V through a step-down circuit based on the LM2596S-ADJ voltage regulator chip design and 7V regulated output, providing a 7V regulated power supply for the micro stepper motor. The power supply of the main control chip achieves a 3.3V regulated output through three-way step-down conversion. Through the voltage stabilization circuit designed based on three voltage stabilization chips LM2596S-ADJ, LM2596S-5.0, and LM2596S-3.3, it realizes 24V to 7V, then from 7V to 5V, and finally from 5V to 3.3V, providing a 3.3V regulated power supply for the main control chip.

5. The fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit according to claim 1, characterized in that: The circuit board also integrates a decoupling circuit, a crystal oscillator circuit, a reset circuit, a start-up circuit, a debugging interface circuit, and a connection circuit. The micro stepping motor is connected to the drive circuit through the connection circuit, and the decoupling circuit, crystal oscillator circuit, reset circuit, start-up circuit, and debugging interface circuit are respectively connected to the main control chip.

6. The fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit according to claim 5, characterized in that: The decoupling circuit is composed of five 100nF chip capacitors in a 0805 package. The decoupling circuit provides a low-impedance power path for the main control chip and filters out transient current noise generated by the switching of logic gates inside the main control chip. The crystal oscillator circuit uses crystal oscillators with frequencies of 8MHz and 32.768kHz. Two identical load capacitors are set at both ends of the crystal oscillator. A load capacitor of 22pF is set at both ends of the 8MHz crystal oscillator, and a load capacitor of 8pF is set at both ends of the 32.768kHz crystal oscillator.

7. The fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit according to claim 5, characterized in that: The reset circuit uses a low-level reset, and a capacitor is connected in parallel with the button for button debounce; when the button is closed, the capacitor is charged and RST is at a high level. When the button is pressed, the capacitor releases current, and the resistor in series with the capacitor will keep the current on the NRST pin from exceeding 3.3V when the switch SW1 is closed; after the button is restored, the capacitor will charge again; during the key restart period, the NRST pin remains at a low level.

8. The fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit according to claim 5, characterized in that: The debug interface circuit is an SWD debug interface circuit. The debug interface circuit reads and writes the registers and memory of the main control chip through the SWDIO pin and SWCLK pin. The SWDIO pin is used for bidirectional data transmission, and the SWCLK pin is used to maintain clock synchronization. The connection circuit uses a 4-pin wire-to-wire connector with a built-in spacing of 2mm.

9. The fully automatic electrochemiluminescence analyzer based on automatic chip entry and exit according to claim 1, characterized in that: The driving circuit is an A4988 driving circuit, which uses two 1×8P connectors; the electrochemiluminescence excitation unit includes an LM2596S-ADJ voltage regulator chip, which uses the LM2596S-ADJ voltage regulator chip and an adjustable potentiometer to achieve a 1.2V-24V wide range adjustable regulated power supply output.

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