CN116539871B - Microfluidic-based chemiluminescence assay device - Google Patents

Abstract

The invention provides a chemiluminescence measurement device, which relates to the field of detection instruments. It includes a device shell, a microfluidic chip module arranged inside the device shell, a reagent module, an automatic pipetting module, an incubation chamber module, a centrifugal motor drive module and a chemical The luminous signal detection module, as well as the data interaction module and power module arranged above the device shell, optimize the structure and position of the device shell and its configured modules, especially the integration of micro-controllers with multiple parallel detection units. The unique structure of the fluid control chip module, the automatic pipetting module driven by 3 sets of motors to move in three directions, the centrifugal motor drive module with both a centrifugal motor and an external magnet drive motor, and the closeable incubation chamber module makes the instrument small in size. , low cost, simple structure, simple and flexible operation, can achieve full automation, and is especially suitable for promotion and application in the field of POCT.

Description

Microfluidic-based chemiluminescence assay device

Technical field

The invention relates to the field of detection instruments, and in particular to a chemiluminescence measurement device, especially a portable, automated, parallel, multi-index joint detection and low-cost chemiluminescence measurement device based on a microfluidic chip for on-site rapid detection.

Background technique

Chemiluminescence immunoassay (CLIA) is a detection and analysis technology that combines highly sensitive chemiluminescence assay technology with highly specific immune reactions. Among them, magnet chemiluminescence immunoassay combines magnetic separation technology and chemiluminescence. An analysis method that combines immunoassay technology with immunoassay technology and is widely used in infectious disease detection, cardiovascular and cerebrovascular disease monitoring, tumor marker detection, and hormone and drug detection.

The centrifugal microfluidic chip can combine the valves, fluid channels, heaters, etc. involved in the processes of sampling, injection, incubation, adsorption and separation of magnetic beads, mixing and detection in the process of magnet chemiluminescence immunoassay. Structures such as separation devices and detectors are integrated on the chip, and centrifugal force is used as the fluid driving force to achieve detection and analysis of multiple indicators of the same sample to be tested or the same indicator of multiple samples to be tested.

Currently, large and medium-sized chemiluminescence detectors on the market have limitations such as large size, complex equipment, cumbersome operation, high equipment cost, and high requirements for the use environment. They often need to be equipped in the testing centers of large and medium-sized hospitals, making it difficult to adapt to primary care. The testing environment and objective needs of the site. Point-of-care testing (POCT), with its characteristics of convenient operation, low cost and short testing time, has been increasingly used in clinical departments, rapid diagnosis, dynamic monitoring, and treatment effect evaluation of critically ill patients. The more widely recognized it is, but at the same time, test accuracy is the lifeblood of whether POCT equipment can become the basis for clinical diagnosis. The advantage of chemiluminescence is its extremely high precision, and microfluidics provides an extremely favorable technical platform for the application of chemiluminescence technology in POCT. Therefore, how to provide a microfluidic-based chemiluminescence measurement device that can take into account the advantages of inspection accuracy and small volume for flexible on-site detection, and can achieve precise control, mixing, reaction and separation of liquids in the microfluidic chip, has become an urgent problem to be solved.

Contents of the invention

The object of the present invention is to at least partially overcome the technical defects of existing chemiluminescence measurement devices in POCT applications, and provide a microfluidic-based chemiluminescence measurement that is centrifugable, miniaturized, simple to operate, low-cost and has high detection accuracy. device.

In order to achieve the above object or one of the objectives, the present invention provides the following technical solutions:

A chemiluminescence measurement device, including a device shell, and a microfluidic chip module, a reagent module, an automatic pipetting module, an incubation chamber module and a centrifugal motor drive module arranged inside the device shell; wherein, ① the microfluidic chip module includes A microfluidic chip and a magnetic bead control layer are provided. The microfluidic chip and the magnetic bead control layer both include a three-layer structure of a cover plate, an intermediate layer and a bottom plate. These three-layer structures are all arranged axially symmetrically. The microfluidic chip is configured There are multiple sets of parallel detection units with the same structure; the magnetic bead control layer is close to the lower surface of the microfluidic chip, including an internal magnet set in an arc-shaped magnet channel; ② The reagent module includes a gun tip, a reagent tube and a wash The liquid test tubes are each arranged in multiple rows arranged in parallel. The number of parallel arrangements is consistent with the number of parallel detection units on the microfluidic chip; ③ The automatic pipetting module is driven by 3 sets of motors to control the front and rear, left and right and up and down All-round and multi-angle pipetting movement, including horizontal left and right movement controlled by the X-axis stepper motor drive; horizontal forward and backward movement controlled by the Y-axis stepper motor drive; and vertical up and down movement controlled by the Z-axis stepper motor drive. ; The automatic pipetting module is fixed and supported above the reagent module through a support column; ④ The incubation chamber module uses an aluminum block heating method, including a slide motor and a heating aluminum block. It is microfluidic by setting a heating upper cover and a sealed lower cover. The chip constructs a closed, light-shielded incubation reaction chamber; ⑤ The centrifugal motor drive module includes a centrifugal motor, a centrifugal motor bracket, a flange base located inside the sealed lower cover, and an external magnet and an external magnet fixing member located outside the sealed lower cover. and an external magnet drive motor; the external magnet drive motor is fixed on the support column and is used to drive the external magnet to magnetically control the internal magnet corresponding to the position.

Preferably, the number of support columns may be a multiple of 2, preferably 4.

Preferably, the external magnet drive motor can be fixed to the support column by pins and screws.

In the present invention, the three-layer structure of the microfluidic chip is the microfluidic chip cover plate, the microfluidic chip middle layer and the microfluidic chip base plate; the three-layer structure of the magnetic bead control layer is the magnetic bead control layer cover. board, magnetic bead control layer middle layer and magnetic bead control layer bottom plate; the three-layer structure of the microfluidic chip is provided with corresponding bonding positioning holes and/or pin holes, and the three-layer structure is connected by bonding positioning holes , and/or secure via pin holes.

Preferably, the three-layer structure is bonded and fixed by hot pressing through bonding positioning holes, and/or fixed with pins through pin holes.

In the present invention, each group of parallel detection units in the middle layer of the microfluidic chip includes a reaction chamber, a buffer chamber, a detection chamber and a waste liquid chamber in sequence; the reaction chamber is connected to the detection chamber through the first capillary valve, and the buffer chamber passes through the buffer chamber. The liquid channel is connected to the detection chamber, and the detection chamber is connected to the waste liquid chamber through a second capillary valve with a circular blocking valve; the reaction chamber is provided with a reaction chamber pore channel, the buffer chamber is provided with a buffer cavity pore channel, and the detection chamber is provided with The detection chamber has a pore channel; the reaction chamber is provided with a reaction chamber sample adding channel, and the buffer chamber is provided with a buffer chamber sample adding channel. The buffer cavity sample adding channel is used to add excitation liquid into the buffer cavity, and under the centrifugal force of the centrifugal motor, it is thrown into the detection cavity through the buffer cavity liquid channel for luminescence reaction.

Preferably, the number of parallel detection units is set to three groups.

Further preferably, the microfluidic chip is disposable.

In the present invention, the middle layer of the magnetic bead control layer is also provided with pin holes, light guide holes and spherical magnetic beads; the spherical magnetic beads are provided at the left and right ends of the arc-shaped magnet channel.

Preferably, one end of the arc-shaped magnet channel corresponds vertically to the position of the reaction chamber.

Further preferably, the number of spherical magnetic beads provided at each end is preferably two.

In the present invention, the distance from the arc-shaped magnet channel to the center of the middle layer of the magnetic bead control layer is consistent with the distance from the reaction chamber to the center of the middle layer of the microfluidic chip.

Preferably, driven by the external magnet driving motor, the magnetic bead control layer rotates counterclockwise, and the internal magnet moves in the arc-shaped magnet channel to the proximal end directly below the reaction chamber to fix the magnetic beads in the reaction chamber; in the external magnet Driven by the drive motor, the magnetic bead control layer rotates clockwise, and the internal magnet moves in the arc-shaped magnet channel to the far end away from the reaction chamber, releasing the magnetic attraction and fixation of the magnetic beads in the reaction chamber, so that the magnetic beads are in the reaction chamber. Free movement inside.

In the present invention, the internal magnet is arranged on the middle layer of the magnetic bead control layer.

According to a preferred embodiment of the present invention, the microfluidic chip module is fixed on the flange base above the centrifugal motor, and the flange base is nested on the central axis of the centrifugal motor above the centrifugal motor bracket. It drives the components fixed on the flange base to rotate and centrifuge around the central axis of the centrifugal motor.

Preferably, the microfluidic chip module is fixed on the flange base above the centrifugal motor through butterfly screws and chip pressing parts.

According to a preferred embodiment of the present invention, in the non-centrifugal state, the external magnet extends into the bottom of the internal magnet inside the sealed lower cover through the external magnet channel provided on the side of the sealed lower cover, and the magnetic force controls the internal magnet in the arc-shaped magnet channel. move.

In the present invention, the slide motor controls the upper heating cover to move up and down in the Z-axis direction to adjust the height of the upper heating cover.

Preferably, the slide motor can heat the height of the upper cover within a range of 50 mm.

In the present invention, the device shell is made of metal material with thermal conductivity. The device shell is provided with a heat sink on the rear side plate, and a heat sink fan is fixed in the middle of the heat sink.

According to a preferred embodiment of the present invention, a data interaction module and a power module are also provided above the device casing. The data interaction module provides a human-computer interaction interface through the touch screen; the power module cooperates with the power adapter to provide stable output power.

Preferably, the power module can provide stable output power of up to 220 watts in conjunction with the power adapter.

Preferably, on the device shell, the data interaction module displays the numerical value or numerical curve of the detected chemiluminescence signal in real time through the touch screen; the data interaction module is also equipped with data processing software to convert the chemiluminescence signal detected from the photomultiplier tube to The concentration of the sample to be tested added to the microfluidic chip is calculated through the calculation of the four-parameter model algorithm or other algorithms, and the concentration data is displayed in real time through the touch screen, thereby achieving quantitative detection of the chemiluminescence signal.

In the present invention, in the incubation chamber module, the slide motor is fixed on the device bottom plate of the device shell through the slide motor support frame, the heating aluminum block is fixed on the inside of the heating upper cover, and the upper surface of the heating aluminum block is closely adhered to the heating film. The heating film corresponds to the center position of the heating aluminum block; the heating aluminum block moves to the top of the microfluidic chip with the heating upper cover to heat the microfluidic chip and control the temperature of the incubation chamber module.

Preferably, the heating aluminum block can be fixed on the inside of the heating upper cover through screws.

Preferably, the heating aluminum block can control the temperature of the incubation chamber module through a PID algorithm.

In the present invention, the chemiluminescence measurement device is also provided with a chemiluminescence signal detection module, which includes a photomultiplier tube and a light guide tube. One end of the light guide tube is fastened to the window of the photomultiplier tube and fixed on the bottom plate of the device shell; The other end of the light guide extends into the bottom of the microfluidic chip module through the light guide channel provided on the surface of the sealing lower cover, and is aligned with the detection cavity of the microfluidic chip for detecting the chemiluminescence signal generated by the microfluidic chip.

In the present invention, the light guide tube is cylindrical and can be composed of an optical fiber (Polymethyl Methacrylate, PMMA) wrapped in a jacket (PVC).

According to a preferred embodiment of the present invention, a grating plate is provided on the upper part of the centrifugal motor, and a slit is provided on the grating plate, and the grating plate is fixed on the side of the flange base; a positioning optical coupler is provided at the bottom of the sealing lower cover, and the positioning optical coupler is Equipped with light channel.

Preferably, the grating sheet can be fixed on the side of the flange base through screws.

Preferably, the slit corresponds to the light channel and is used to position the microfluidic chip.

Preferably, there is one slit on the grating sheet, and the slit width is 0.1 mm-0.5 mm, for example, it can be 0.1 mm, 0.2 mm, 0.3 mm, 0.4 mm or 0.5 mm, etc.

In the present invention, in the centrifugal motor drive module, the centrifugal motor can be a servo motor, a stepper motor, a brushless DC motor or other motors, and can provide a rotational speed in the range of 1000 rpm-12000 rpm. The rotational speed can be, for example, 1000rpm, 2000rpm. , 3000 rpm, 4000 rpm, 5000 rpm, 6000 rpm, 7000 rpm, 8000 rpm, 9000 rpm, 10000 rpm, 11000 rpm or 12000 rpm, etc.

In the present invention, in the automatic pipetting module, three sets of motors control the number of steps the motor moves by converting electrical pulse signals into corresponding linear displacements; the movement in the X-axis and Y-axis directions passes through the X-axis guide rail and Y-axis guide rail respectively. To realize, the movement in the Z-axis direction is realized by the Z-axis motor.

Preferably, the Z-axis motor is a screw motor.

Further preferably, three corresponding touch switches are provided for the three sets of motors in the three directions of the X-axis, Y-axis and Z-axis, namely the X-axis touch switch, the Y-axis touch switch and the Z-axis touch switch.

Further preferably, in the Z-axis direction, one end of the pipette body is fixed with a pipette, and the other end of the pipette body is fixed with a hose connector. The hose connector is connected to the plunger pump through the air hose, and is connected to the plunger pump through the column. The movement of the plug pump controls the air pressure in the air hose, and then controls the pipetting action of the pipette body through the air pressure.

Further preferably, the gas line hose is a flexible, transparent pipe.

In the present invention, the chemiluminescence measurement device is also provided with an auxiliary module. The auxiliary module includes a main control board and a main control board bracket. The main control board is fixed on the main control board bracket. The main control board bracket is fixed on the device bottom plate of the device shell. The main control board is arranged parallel to the rear side panel of the device shell; the main control board controls the operation of other modules and provides data interaction with the data interaction module.

In the present invention, the reagent module also includes a reagent kit, a reagent kit support plate and a reagent kit support frame. The reagent kit is fixed on the reagent kit support plate through the protruding latching structure of the reagent kit support plate. The reagent kit support frame is connected to the device through screws. The device bottom plate of the shell is fixed; the test box is provided with holes for placing the gun tip, reagent test tubes and washing liquid test tubes.

Preferably, the kit is also provided with a tip waste bin.

Further preferably, corresponding to the number of three groups of parallel detection units in the middle layer of the microfluidic chip, the gun tips, reagent test tubes and washing liquid test tubes are arranged in parallel in three rows, and are loaded onto the microfluidic chip respectively. 3 sets of parallel detection units.

The present invention also claims the application of the above chemiluminescence assay device in chemiluminescence immunoassay.

Beneficial effects of the present invention:

The chemiluminescence assay device of the present invention includes a device shell, a microfluidic chip module, a reagent module, an automatic pipetting module, an incubation chamber module and a centrifugal motor drive module arranged inside the device shell. In addition, it is also provided with A chemiluminescence signal detection module using a high-sensitivity photomultiplier tube and an auxiliary device using a main control board to control the operation of other modules. Among them, the device shell is made of metal material with good thermal conductivity, which can quickly and effectively conduct away the heat from the centrifugal motor driver that is close to the inside of the rear side panel. There are also heat sinks on the rear side panel, and a heat sink fan is nested in the middle of the heat sink, which further speeds up the absorption and conduction of excess heat. At the same time, partitions are provided in the centrifugal motor driver and incubation chamber module inside the device to effectively prevent the heat generated by the centrifugal motor driver from affecting the control of the reaction temperature during the reaction process, thereby achieving accurate temperature control.

A data interaction module and a power module are also provided above the device shell. The power module, combined with the power adapter, can provide up to 220 watts of output power, providing stable high-power output for the device to ensure normal and smooth operation of the device. The data interaction module provides a human-computer interaction interface through the touch screen. After receiving relevant instructions, the human-computer interaction interface realizes automatic control of the chemiluminescence detection device, information collection, storage, and display through the main control board of the auxiliary module. , analysis and feedback functions. In addition, the data interaction module displays the numerical value or numerical curve of the detected chemiluminescence signal in real time through the touch screen; the data processing software configured in the data interaction module can process the chemiluminescence signal detected from the photomultiplier tube through a four-parameter model algorithm. Or other algorithms are used to calculate the concentration of the sample to be tested added to the microfluidic chip, and the detection results of the concentration data are displayed in real time through the touch screen, thereby achieving real-time quantitative detection of the chemiluminescence signal.

The microfluidic chip module of the present invention is composed of a microfluidic chip with a three-layer structure and a magnetic bead control layer. The three-layer structure is arranged axially symmetrically. The microfluidic chip can be equipped with multiple groups of parallel detection units with the same structure, which can realize Joint detection of multiple markers for the same sample to be tested, or joint detection of the same marker for multiple samples to be tested, to achieve rapid parallel detection.

For example, when there are three groups of parallel detection units, it can be set up as a microfluidic chip targeting three myocardial necrosis markers related to myocardial infarction. The three myocardial necrosis markers can include: troponin (cTnI), creatine kinase Isoenzyme (CK-MB) and myoglobin (Myo) realize rapid joint detection and minimize the detection time. This enables instant diagnosis and timely treatment of myocardial infarction, and rapid detection of critical diseases such as myocardial infarction. and timely diagnosis, which has important clinical application value and significance.

In addition, each group of parallel detection units in the middle layer of the microfluidic chip includes a reaction chamber, a buffer chamber, a detection chamber and a waste liquid chamber in sequence. The reaction chamber is connected to the detection chamber through the first capillary valve, and the buffer chamber passes through the buffer chamber liquid channel. It is connected to the detection chamber, and the detection chamber is connected to the waste liquid chamber through a second capillary valve with a circular blocking valve; the first capillary valve, the second capillary valve and the circular blocking valve prevent the sample or reagent in the chamber from being Before the reaction is completed, it is thrown into the next chamber. Among them, the first capillary valve connects the reaction chamber and the detection chamber to prevent magnetic beads, samples or reagents from being thrown into the detection chamber when the reaction chamber is mixed, and avoids uneven mixing of some magnetic beads; the second capillary valve connects the detection chamber cavity and waste liquid cavity to prevent the magnetic beads and pre-excitation liquid from being thrown directly into the waste liquid cavity when they are transferred to the detection cavity, thereby avoiding the loss of magnetic beads; the circular blocking valve is set in the middle of the second capillary valve. Further increase the breakthrough speed of the second capillary valve to further avoid the loss of magnetic beads.

The microfluidic chip can also be driven by a centrifugal motor to generate Euler force or centrifugal force. Under the action of Euler force or centrifugal force, the magnetic beads, samples or reagents complete the various steps of the chemiluminescence reaction in each chamber in sequence: first, in The magnetic bead solution and sample are added to the reaction chamber, and the two are mixed under the action of Euler force. After the mixing is completed, the magnetic beads are adsorbed. The solution and sample quickly pass through the detection chamber under the action of centrifugal force, and are finally thrown into the waste liquid. into the chamber; secondly, the magnetic beads are cleaned by passing the washing solution three times. The cleaned waste liquid passes through the detection chamber and is thrown into the waste liquid chamber under the action of centrifugal force; after that, a luminescent marker is added to the reaction chamber. , mix with the magnetic beads again, and pass in the washing liquid three times to clean the magnetic beads. The waste liquid generated in this process is also thrown into the waste liquid chamber; after that, the pre-excitation liquid is added to the reaction chamber. They are transferred to the detection chamber together with the magnetic beads. Finally, the excitation fluid is added to the buffer chamber and thrown into the detection chamber under the action of centrifugal force. The magnetic beads, pre-excitation fluid and excitation fluid react and emit light in the detection chamber, completing the process. Chemiluminescence reaction experiment.

In the chemiluminescence measurement device of the present invention, the internal magnet is arranged on the middle layer of the magnetic bead control layer, and the optimization strategy of rapid switching of the working state is realized through the movement of the internal magnet, that is, through the real-time control and rapid switching of the position of the internal magnet, thereby achieving Different reaction operations such as magnetic bead adsorption, aggregation or mixing. Specifically, the magnetic bead control layer is placed close to the lower surface of the microfluidic chip, in which an arc-shaped magnet channel is designed. The distance from the arc-shaped magnet channel to the center of the middle layer of the magnetic bead control layer and the distance from the reaction chamber to the microfluidic chip The distance between the layer centers is consistent, and one end of the arc-shaped magnet channel corresponds to the reaction chamber of the microfluidic chip up and down, and the internal magnet is arranged in the arc-shaped magnet channel. The internal magnet is located at one end of the arc-shaped magnet channel away from the reaction chamber of the microfluidic chip, which is called the distal end. Correspondingly, when the internal magnet is located at the end directly below the reaction chamber of the microfluidic chip, it is called the near end. end. During the reaction process, when the reagent needs to be transferred through the high-speed rotation of the microfluidic chip or the waste liquid passes through the detection chamber and is finally thrown into the waste liquid chamber, in order to prevent the magnetic beads from being thrown to the next chamber, an internal magnet is required to align the magnetic beads. At this time, the external magnet, driven by the external magnet drive motor, first extends horizontally under the magnetic bead control layer. The magnetic bead control layer rotates counterclockwise. The internal magnet in the arc-shaped magnet channel will move under the external magnet. Under the influence of magnetic force, it automatically moves along the arc-shaped magnet channel to the proximal end (that is, directly below the reaction chamber), and efficiently adsorbs and fixes the magnetic beads. During the reaction, when the magnetic beads and samples or reagents need to be mixed, the external magnet is driven by the external magnet driving motor and extends horizontally under the magnetic bead control layer. The magnetic bead control layer rotates clockwise, and the internal magnet will move outside. Under the influence of the magnetic force of the magnet, it automatically moves along the arc-shaped magnet channel to the far end (that is, away from the reaction chamber) without adsorbing the magnetic beads, allowing free movement and mixing of the magnetic beads.

The reagent module of the present invention corresponds to the number of parallel detection units in the middle layer of the microfluidic chip. The gun tips, reagent test tubes and washing liquid test tubes are arranged in parallel in the same number of rows, and samples are added to the microfluidic chip respectively. of each detection unit. For example, when there are three groups of parallel detection units in the middle layer of the microfluidic chip, the gun tips, reagent test tubes, and wash solution test tubes are arranged in parallel in three rows, and samples can be added to the three detection units on the microfluidic chip. . In addition, the kit is also equipped with a tip waste bin, which is used to store discarded tip tips after each sample addition to avoid contamination.

The automatic pipetting module of the present invention is equipped with three sets of motors in three directions. The three sets of motors control the number of steps the motor moves by converting electrical pulse signals into corresponding linear displacements, thereby realizing the movement of the pipette in the X-axis, Y-axis. Precise movement in three directions: the X-axis and the Y-axis are realized by guide rails, and the movement of the pipette in the Z-axis direction is realized by the Z-axis motor. The Z-axis motor is preferably a screw motor. In addition, touch switches are provided in the X, Y, and Z axes to determine the zero point position of movement, and can flexibly and conveniently realize operations such as loading and dropping of the gun tip, and sampling and injection of samples. One end of the pipette body is fixed with a pipette to load and remove the gun tip; the other end of the pipette body is fixed with a hose connector, which is connected to the plunger pump through the air hose, and is connected to the plunger pump through the column. The movement of the stopper pump causes the air pressure in the air hose to increase or decrease, thereby controlling the sampling and injection of samples or reagents through air pressure, that is, controlling the pipetting action through air pressure; among them, the air line soft The tube is a flexible transparent hose. Due to the three sets of motors in three directions, the sample adding position of the automatic pipetting module can be fixed, improving the accuracy of pipetting operations.

In the centrifugal motor drive module of the present invention, the centrifugal motor can be a servo motor, a stepper motor, a DC brushless motor or other motors, and can provide a rotational speed in the range of 1000 rpm-12000 rpm to meet the various reaction steps required for the chemiluminescence reaction. Requires rotational speed; with the help of the centrifugal force generated by high-speed rotation, combined with the functional design of the reaction chamber and channel on the microfluidic chip, the sample or reaction reagent to be tested can gradually complete the various reaction steps required for the chemiluminescence reaction on the microfluidic chip. For example, there are several steps such as mixing the sample and magnetic beads, cleaning the magnetic beads, transferring reagents, and waste disposal. A grating plate is fixed on the centrifugal motor, and a slit is provided on the grating plate. A positioning optical coupler is fixed at the bottom of the sealed lower cover. The positioning optical coupler is provided with an optical channel. The slit corresponds to the optical channel of the positioning optical coupler. During the rotation of the fluidic chip, the slits will continuously pass through the positioning optical coupler to position the microfluidic chip.

The incubation chamber module of the present invention is specially equipped with a heating upper cover and a sealing lower cover. By moving the heating upper cover up and down, an incubation chamber with dual functions can be constructed together with the sealing lower cover. On the one hand, it can provide a closed temperature control environment to achieve accurate temperature control of the microfluidic chip module and overcome the interference and influence of ambient temperature fluctuations; on the other hand, it can provide a light-shielding environment to overcome the impact of ambient stray light on high-sensitivity chemiluminescence signal collection based on photomultiplier tubes. interference and influence to achieve accurate chemiluminescence signal collection and analysis. Furthermore, the slide motor controls the movement of the heating upper cover in the vertical direction through the heating upper cover connecting piece. The height of the heating upper cover can be adjusted within a range of 50 mm, so that the heating upper cover is in a lifted state before placing the chip. It will not affect the placement of the microfluidic chip module. After placing the microfluidic chip, the heating upper cover falls and is tightly fastened to the sealed lower cover to form an incubation chamber.

An external magnet driving motor is also provided below the incubation chamber module of the present invention. The external magnet driving motor is fixed on the support column and is used to drive the external magnet. The external magnet can extend into the internal magnet through the external magnet channel on the right side of the sealed lower cover. Below, the internal magnet can be moved from the proximal end to the distal end or from the distal end to the proximal end. The structure is simple, and at the same time, the working state of the internal magnet is switched flexibly, which greatly improves the continuity and flexibility of the response, and at the same time directly Fixed on the support column, the instrument structure is optimized and the layout of the internal parts of the instrument is more compact.

The incubation chamber module of the present invention is integrated into the chemiluminescence measurement device, which effectively improves the comprehensive performance of the measurement device. The entire incubation chamber module is temperature controlled through the PID algorithm, and the incubation time is adjusted in time according to the temperature setting value required for the chemiluminescence reaction. The temperature in the chamber is dynamically adjusted so that the temperature in the entire incubation chamber is always maintained within the temperature required for the reaction, ensuring the accuracy of the final chemiluminescence signal detection. At the same time, a capsule-shaped through structure is designed at the same position of the heating upper cover, heating film and heating aluminum block, which can minimize the light leakage area of the microfluidic chip. The heating aluminum block is fixed on the inside of the heating upper cover, and the heating film is close to the upper surface of the heating aluminum block, further providing a stable temperature for the corresponding heating aluminum block.

In the luminescent signal detection module of the present invention, a light guide is specially provided. One end is tightly aligned with the window of the photomultiplier tube, and the other end extends into the bottom of the microfluidic chip module through the light guide channel of the sealed lower cover. Surface, align it with the detection chamber of the microfluidic chip. The light guide not only solves the problem that the photomultiplier tube cannot be placed directly under the microfluidic chip module to detect chemiluminescence signals, but it can also quickly collect and transmit the weak chemiluminescence signals generated by the reaction, preventing the weak luminescence signals from being transmitted inside the instrument. The loss in space improves the accuracy of detection; and the diameter of the light guide hole is basically the same as the diameter of the light guide, effectively overcoming the interference and influence of ambient stray light on the collection of high-sensitivity chemiluminescence signals based on photomultiplier tubes. At the same time, the luminescence signal can be collected quickly and accurately. The auxiliary device of the present invention controls the operation of other modules through the main control board, where the main control board not only controls the operation of other modules, but also provides data interaction with the data interaction module. When the magnetic beads, pre-excitation solution and excitation solution enter the detection chamber and react to emit light, the weak light signal generated will be transmitted to the photomultiplier tube through the light guide. The photomultiplier tube can amplify the weak light signal and convert it into an electrical signal. , thereby realizing the collection of chemiluminescence signals and the reaction luminescence of magnetic beads, pre-excitation liquid and excitation liquid in the detection cavity, amplifying the weak light signal that may be lost during the detection process, further improving the accuracy of detection.

Compared with the large and medium-sized chemiluminescence detectors currently on the market, the chemiluminescence measurement device of the present invention combines the advantages of chemiluminescence, microfluidics and POCT, and is centrifugable, compact, easy to move, simple to operate, and highly automated. It has the characteristics of high, low equipment cost, short detection time, flexible use and high detection accuracy. It is an instant chemiluminescence detection system that is more suitable for on-site rapid detection (POCT) application scenarios. At the same time, with the help of the disposable microfluidic of the present invention The cooperation between chips and detection equipment is more suitable for rapid multi-index or multi-item joint detection of POCT in primary medical units, such as multi-index joint and rapid detection of critical diseases such as myocardial infarction, which has important clinical application value and significance. .

Description of the drawings

Figure 1 is a perspective view of the right front side of the chemiluminescence measurement device of the present invention;

Figure 2 is a perspective view of the left rear side of the chemiluminescence measurement device of the present invention;

Figure 3 is a three-dimensional view of the structure of the microfluidic chip and magnetic bead control layer of the chemiluminescence measurement device of the present invention;

Figure 4 is a top view of the microfluidic chip cover of the chemiluminescence assay device of the present invention;

Figure 5 is a top view of the microfluidic chip base plate of the chemiluminescence assay device of the present invention;

Figure 6 is a top view of the middle layer of the microfluidic chip of the chemiluminescence measurement device of the present invention;

Figure 7 is a top view of the magnetic bead control layer cover plate and the magnetic bead control layer bottom plate of the chemiluminescence measurement device of the present invention;

Figure 8 is a top view of the middle layer of the magnetic bead control layer of the chemiluminescence measurement device of the present invention;

Figure 9 is an enlarged view of the partial structure of the middle layer of the magnetic bead control layer of the chemiluminescence measurement device of the present invention;

Figure 10 is a perspective view of the left front side of the internal structure of the device housing of the chemiluminescence measurement device of the present invention;

Figure 11 is a perspective view of the rear right side of the internal structure of the device housing of the chemiluminescence measurement device of the present invention;

Figure 12 is a perspective view of the front left side of the reagent module of the chemiluminescence measurement device of the present invention;

Figure 13 is a perspective view of the left rear side of the X-axis part of the automatic pipetting module of the chemiluminescence assay device of the present invention;

Figure 14 is a perspective view of the front right side of the X-axis part of the automatic pipetting module of the chemiluminescence assay device of the present invention;

Figure 15 is a side perspective view of the Y-axis part of the automatic pipetting module of the chemiluminescence assay device of the present invention;

Figure 16 is a side perspective view of the Z-axis part of the automatic pipetting module of the chemiluminescence assay device of the present invention;

Figure 17 is a side perspective view of the incubation chamber module of the chemiluminescence assay device of the present invention;

Figure 18 is a bottom view and a perspective view of the heated upper cover of the incubation chamber module of the chemiluminescence measurement device of the present invention;

Figure 19 is a perspective view and a bottom view of the sealed lower cover of the incubation chamber module of the chemiluminescence measurement device of the present invention;

Figure 20 is a front view of the chemiluminescence signal detection module of the chemiluminescence measurement device of the present invention;

Figure 21 is a perspective view of the centrifugal motor drive module of the chemiluminescence measurement device of the present invention;

Figure 22 is a perspective view of the external magnet driving motor of the chemiluminescence measurement device of the present invention.

In the picture: 1-Reaction chamber sample hole; 2-Buffer chamber air hole; 3-Buffer chamber sample hole; 4-Reaction chamber air hole; 5-Detection chamber air hole; 6-Bonding positioning hole; 7-Pin hole; 8 -Reaction chamber; 9-Reaction chamber sample addition channel; 10-Buffer chamber pore channel; 11-Buffer chamber sample addition channel; 12-Buffer chamber; 13-Buffer chamber liquid channel; 14-Detection chamber; 15-Circular blocking Valve; 16-second capillary valve; 17-waste liquid chamber; 18-reaction chamber pore channel; 19-first capillary valve; 20-detection chamber pore channel; 21-light guide hole; 22-internal magnet; 23-spherical Magnetic beads; 24-microfluidic chip cover; 25-microfluidic chip middle layer; 26-microfluidic chip bottom plate; 27-magnetic bead control layer cover; 28-magnetic bead control layer middle layer; 29-magnetic beads Bead control layer bottom plate; 30-touch screen; 31-upper side panel; 32-front panel; 33-hatch handle; 34-hatch door; 35-rubber foot pad; 36-hinge; 37-right side panel; 38-right side panel handle; 39-rear side panel; 40-heat sink; 41-heat sink fan; 42-left side panel; 43-left side panel handle; 44-automatic pipetting module; 45-reagent module; 46 -Stepper motor driver; 47-Device base plate; 48-Chemiluminescence signal detection module; 49-Centrifugal motor drive module; 50-Incubation chamber module; 51-Microfluidic chip module; 52-Main control board; 53-Main control Plate bracket; 54-centrifugal motor driver; 55-piston pump fixing piece; 56-piston pump solenoid valve fixing piece; 57-piston pump solenoid valve; 58-piston pump; 59-centrifugal motor driver partition; 60 -Driving shaft bearing; 61-reagent kit; 62-reagent kit support plate; 63-reagent kit support frame; 64-latching structure; 65-gun tip waste bin; 66-reagent test tube; 67-wash solution test tube; 68- Gun tip; 69-X-axis left driving wheel; 70-X-axis touch switch holder; 71-X-axis touch switch; 72-X-axis left gear plate fixing piece; 73-X-axis stepper motor; 74-X Axis left gear plate; 75-X-axis drag chain; 76-X-axis left support plate; 77-X-axis left driven wheel fixing; 78-X-axis left driven wheel; 79-support column; 80-X-axis right cantilever pin ; 81-X-axis guide rail; 82-X-axis guide rail slider; 83-Y-axis support plate; 84-X-axis left cantilever pin; 85-X-axis right driven wheel fixed piece; 86-X-axis right driven wheel; 87- X-axis right support plate; 88-X-axis belt; 89-X-axis right tooth plate fixing piece; 90-X-axis right tooth plate; 91-X-axis right driving wheel; 92-X-axis right bearing fixing piece; 93-Drive Axis; 94-X-axis stepper motor synchronous wheel; 95-X-axis stepper motor holder; 96-drive shaft belt; 97-drive shaft synchronous wheel; 98-X-axis left bearing fixation; 99-Y-axis stepper Motor; 100-Y-axis stepper motor bracket; 101-Y-axis driving wheel; 102-Y-axis belt; 103-pipettor base plate; 104-Y-axis tooth plate; 105-Y-axis guide rail frame; 106-Y-axis slave Moving wheel; 107-Y-axis driven wheel fixing; 108-Y-axis cantilever pin; 109-Y-axis touch switch; 110-Y-axis touch switch holder; 111-Y-axis drag chain bottom plate; 112-Y-axis guide rail slide block; 113-pipettor bottom plate block; 114-Y-axis drag chain bracket; 115-Y-axis drag chain; 116-Y-axis guide rail; 117-Z-axis motor; 118-Z-axis touch switch; 119-Z-axis Guide shaft; 120-top bracket of pipette slider; 121-pipettor slider; 122-hose connector; 123-pipettor body; 124-pipettor; 125-pipettor bottom plate tail stop ; 126-bottom bracket of pipette slider; 127-Z-axis guide shaft bearing; 128-side connector of pipette slider; 129-spring; 130-Z-axis motor fixing piece; 131-heating upper cover connection Parts; 132-Slide motor support frame; 133-Slide motor; 134-Heated upper cover; 135-Sealed lower cover; 136-Heated aluminum block; 137-Heated film; 138-Light pipe channel; 139-External magnet Channel; 140-positioning optocoupler; 141-positioning optocoupler wiring hole; 142-light guide; 143-photomultiplier tube; 144-butterfly screw; 145-chip clamping piece; 146-microfluidic chip; 147 -Magnetic bead control layer; 148-grating plate; 149-flange base; 150-centrifugal motor; 151-centrifugal motor bracket; 152-external magnet drive motor; 153-external magnet fixing piece; 154-external magnet; 155-DC Motor touch switch fixing; 156-DC motor touch switch; 157-guide rail frame; 158-guide rail bearing; 159-DC motor slider; 160-guide rail; 161-DC motor; 162-DC motor bracket; 163-capsule Shaped through structure; 164-air hose.

Detailed ways

Exemplary embodiments of the present invention are described in detail below with reference to the accompanying drawings, in which the same or similar reference numerals represent the same or similar elements. Additionally, in the following detailed description, for convenience of explanation, numerous specific details are set forth to provide a comprehensive understanding of the disclosed embodiments. It will be apparent, however, that one or more embodiments may be practiced without these specific details. In other instances, well-known structures and devices are illustrated in order to simplify the drawings.

According to the general concept of the present invention, a new chemiluminescence assay device for detecting microfluidic chips is provided. The chemiluminescence assay device includes a device shell, a microfluidic chip module and a reagent module arranged inside the device shell. , automatic pipetting module, incubation chamber module, centrifugal motor drive module and chemiluminescence signal detection module, as well as a data interaction module and a power supply module set above the device shell. The chemiluminescence assay device integrates a microfluidic chip module with multiple parallel detection units, an automatic pipetting module driven by 3 sets of motors to move in 3 directions, and a centrifugal motor drive module that combines a centrifugal motor and an external magnet drive motor. And the unique structure of the sealable incubation chamber module, and the structure of the reagent module matches the number of parallel detection units of the microfluidic chip module, and the gap between the internal magnet of the microfluidic chip module and the external magnet of the centrifugal motor drive module Magnetic force enables real-time control and rapid switching of different reaction states of magnetic beads. The chemiluminescence measurement device of the present invention has the characteristics of small size, convenient movement, simple operation, low equipment cost, short detection time, flexible use, high detection accuracy, etc. It can also realize automatic control and is particularly suitable for promotion and application in the field of POCT.

The present invention will be further described in detail below with reference to Figures 1-22.

Figures 1 and 2 respectively show a perspective view of the front right side and the rear left side of the chemiluminescence measurement device of the present invention. It can be seen from Figure 10 that the chemiluminescence measurement device of the present invention includes a device shell and components arranged inside the device shell. A data interaction module and a power module are also provided above the device shell. The device shell includes an upper side plate 31, a device bottom plate 47, a front panel 32, a rear side plate 39, a left side plate 42 and a right side plate 37; the upper side plate 31 and the device bottom plate 47 are respectively connected to the left side plate 42 and the right side plate through screws. The plate 37, the front panel 32 and the rear side panel 39 are connected to ensure the fastening of the shell; the device shells are all made of metal materials with good thermal conductivity, which can quickly and effectively conduct heat away.

The data interaction module is arranged on the upper side plate 31 of the device housing. A rubber foot pad 35 is provided at the bottom of the device bottom plate 47 so that the device is in soft contact with the placement surface to avoid bumping. The hatch 34 is set on the front panel 32 through the hinge 36 to facilitate the user to place and take out the microfluidic chip module 51; the convex structure on the inside of the hatch 34 corresponds to the concave structure on the front panel 32 to ensure sealing; The hatch 34 is also provided with a hatch handle 33, which is fixed on the hatch 34 with screws to facilitate the user to open and close the hatch 34; the rear side panel 39 is provided with a heat sink 40, and the heat sink 40 A heat sink fan 41 is also fixedly installed in the middle to further prevent the device of the present invention from being interfered and affected by the heat generated by the operation of each component during operation, thereby providing a stable temperature environment; the left side panel 42 and the right side panel are The board 37 is respectively provided with a right panel handle 38 and a left panel handle 43. The left panel handle 43 and the right panel handle 38 are fixed on the left panel 42 and the right panel 37 through screws to facilitate the user to carry the mobile device.

It can also be seen in Figure 1 that the data interaction module includes a touch screen 30. The touch screen 30 is fixed above the outer surface of the upper side plate 31 of the device housing. The data interaction module provides the user with a friendly human-computer interaction interface and transmits corresponding instructions to the assistant. The module performs corresponding actions and receives and processes the corresponding data collected by the auxiliary module. It can realize automatic real-time drawing of the luminescence value curve and display the detection results. Specifically, the data interaction module displays the numerical value or numerical curve of the detected chemiluminescence signal in real time through the touch screen 30; the data interaction module is also configured with data processing software to convert the chemiluminescence signal detected from the photomultiplier tube 143 to Calculation is performed through a four-parameter model algorithm or other algorithms to calculate the concentration of the sample to be tested added to the microfluidic chip 146, and the concentration data is displayed in real time through the touch screen 30, thereby achieving quantitative detection of the chemiluminescence signal.

Figure 3 is a three-dimensional view of the microfluidic chip and magnetic bead control layer structure of the chemiluminescence measurement device of the present invention. As can be seen from Figure 3, the microfluidic chip module 51 of the chemiluminescence measurement device of the present invention includes a microfluidic chip 146 and a magnetic bead control layer. The bead control layer 147 (see Figure 21 for location) and the magnetic bead control layer 147 are placed close to the lower surface of the microfluidic chip 146. The three-layer structure of the microfluidic chip 146 includes a microfluidic chip cover 24, a microfluidic chip middle layer 25 and a microfluidic chip bottom plate 26; the magnetic bead control layer 147 includes a magnetic bead control layer cover 27, a magnetic bead control layer The middle layer 28 and the magnetic bead control layer bottom 29.

Figures 4, 5 and 6 respectively show the three-layer structure of the microfluidic chip cover plate 24, the microfluidic chip bottom plate 26 and the microfluidic chip middle layer 25 of the microfluidic chip module 51 of the present invention. These three-layer structures are all arranged axially symmetrically. Each of these three-layer structures is provided with bonding positioning holes 6 and pin holes 7. The three-layer structures are bonded by hot pressing to ensure that the microfluidic chip 146 is loaded. accuracy.

The microfluidic chip 146 is provided with multiple groups of parallel detection units with the same structure. It can be seen from the figure that the number of parallel detection units arranged axially symmetrically is preferably 3 groups. The following description takes one group of them as an example. Regarding parallel detection units: Setting up multiple groups of parallel detection units can, on the one hand, achieve multi-index joint detection for the same sample to be tested. For example, three parallel detection units with the same structure can be constructed to detect three myocardial necrosis markers related to myocardial infarction. Including: troponin (cTnI), creatine kinase isoenzyme (CK-MB) and myoglobin (Myo) to achieve rapid parallel detection; on the other hand, it can be used to detect multiple different samples to be tested at the same time. Fast parallel detection of indicators.

Figure 6 shows: each group of parallel detection units in the middle layer 25 of the microfluidic chip includes a reaction chamber 8, a buffer chamber 12, a detection chamber 14 and a waste liquid chamber 17 in sequence; the reaction chamber 8 communicates with the detection chamber through the first capillary valve 19 14 is connected, the buffer chamber 12 is connected to the detection chamber 14 through the buffer chamber liquid channel 13, and the detection chamber 14 is connected to the waste liquid chamber 17 through the second capillary valve 16 with a circular blocking valve 15; the reaction chamber 8 is provided with a reaction chamber The pore channel 18, the buffer chamber 12 is provided with the buffer cavity pore channel 10, the detection chamber 14 is provided with the detection cavity pore channel 20; the reaction chamber 8 is provided with a reaction chamber sample addition channel 9, and the buffer chamber 12 is provided with a buffer cavity sample addition channel 11, The buffer chamber sample adding channel 11 is used to add excitation liquid into the buffer chamber 12, and under the centrifugal force of the centrifugal motor 150, it is thrown into the detection chamber 14 through the buffer chamber liquid channel 13 for luminescence reaction.

The first capillary valve 19 can prevent magnetic beads, samples or reagents from being thrown into the detection chamber 14 when the reaction chamber 8 is mixed, thereby avoiding uneven mixing of some magnetic beads; the second capillary valve 16 can prevent the magnetic beads and pre-excitation liquid from being thrown into the detection chamber 14. When transferred to the detection chamber 14, they are directly thrown into the waste liquid chamber 17 to avoid the loss of magnetic beads; the circular blocking valve 15 and the second capillary valve 16 prevent the reagent from entering the waste liquid chamber at a lower speed. Avoid reagent loss. Among them, the circular blocking valve 15 can further prevent the breakthrough rotation speed of the second capillary valve 16, thereby avoiding further loss of magnetic beads or reagents. In addition, the first capillary valve 19, the second capillary valve 16 and the circular blocking valve 15 respectively prevent the samples or reagents in the reaction chamber 8 and the detection chamber 14 from being thrown into the reaction chamber before the end of the reaction. Waste liquid chamber 17.

Specifically, the magnetic bead solution and the sample are injected into the reaction chamber 8 through the reaction chamber sampling channel 9 to perform a mixing reaction. After the mixing is completed, the chip is centrifuged under the action of the centrifugal force (or Euler force) generated by the centrifugal motor 150, and the Solutions other than magnetic beads are thrown into the waste liquid chamber 17 through the first capillary valve 19 and the second capillary valve 16, and then the washing liquid is added from the reaction chamber sampling channel 9, and the magnetic beads are mixed and cleaned three times. After completion, centrifugation is performed under the centrifugal force generated by the centrifugal motor 150, and the washing liquid is thrown into the waste liquid chamber 17 through the first capillary valve 19 and the second capillary valve 16, and then the luminescent marker is added from the reaction chamber sampling channel 9 The materials are mixed and reacted. After three times of mixing, centrifugation is performed at a high speed generated by the centrifugal motor, and the luminescent marker waste liquid except the magnetic beads is thrown into the waste liquid through the first capillary valve 19 and the second capillary valve 16. In the liquid chamber 17, add the washing liquid from the reaction chamber sampling channel 9, mix and clean the magnetic beads three times, centrifuge at the high speed generated by the centrifuge motor 150, and pass through the first capillary valve 19 and the second capillary valve 16 Throw the washing liquid into the waste liquid chamber 17, and then add the pre-excitation liquid from the reaction chamber sample addition channel 9. The pre-excitation liquid and the magnetic beads in the reaction chamber are transferred to the detection chamber 14 by the centrifugal force generated by the rotation of the centrifuge motor 150. , then add the excitation liquid from the buffer chamber sample adding channel 11 and throw it into the detection chamber 14 to emit light.

Regarding the centrifugal effect in the above reaction: driven by the centrifugal motor 150, the microfluidic chip 146 generates Euler force or centrifugal force, and the magnetic beads, samples or reagents sequentially complete chemiluminescence in each chamber under the action of Euler force or centrifugal force. Various steps of the reaction: First, add the magnetic bead solution and sample to the reaction chamber 8, and the two are mixed under the action of Euler force. After the mixing is completed, the magnetic beads are adsorbed, and the solution and sample are finally thrown in under the action of centrifugal force. to the waste liquid chamber 17; secondly, the magnetic beads are cleaned by passing the washing liquid three times. The cleaned waste liquid is finally thrown into the waste liquid chamber 17 under the action of centrifugal force; then, add The luminescent markers are mixed with the magnetic beads again, and the magnetic beads are washed by passing washing liquid three times. The waste liquid generated in this process is finally thrown into the waste liquid chamber 17; after that, in the reaction chamber 8 Add the pre-excitation liquid and transfer it to the detection chamber 14 together with the magnetic beads; finally, add the excitation liquid to the buffer chamber 12 and throw it into the detection chamber 14 under the action of centrifugal force. The magnetic beads, pre-excitation liquid and excitation liquid are three. They jointly perform a luminescence reaction in the detection chamber 14 to complete the chemiluminescence reaction experiment.

In Figure 6, since both the microfluidic chip cover plate 24 and the microfluidic chip base plate 26 are provided with air holes at corresponding positions of the air hole channels, the reaction chamber 8 located on the middle layer 25 of the microfluidic chip is provided with reaction chamber air hole channels 18, The buffer chamber 12 is provided with a buffer chamber pore channel 10 and the detection chamber 14 is provided with a detection chamber pore channel 20, which communicate with the internal and external atmospheric pressure of the microfluidic chip 146 through the reaction chamber pore 5, the buffer chamber pore 2 and the detection chamber pore 4 respectively, so that Smooth sample addition.

Figures 7, 8 and 9 respectively show a top view of the magnetic bead control layer cover plate 27 and the magnetic bead control layer bottom plate 29, a top view of the magnetic bead control layer middle layer 28 and the magnetic bead control layer in the microfluidic chip module 51 of the present invention. An enlarged view of the local structure of the position of the spherical magnetic beads 23 in the middle layer 28. It can be seen from Figure 7 that the magnetic bead control layer cover 27 and the magnetic bead control layer bottom plate 29 have the same structure, and both include light guide holes 21 and pin holes 7. The positions of three light guide holes 21 correspond to the microfluidic chip 146 respectively. The position of the detection cavity 14 facilitates the detection of the luminescent signal. Figures 8 and 9 both involve the magnetic bead control layer middle layer 28. It can be seen from Figure 8 that corresponding to the number of parallel detection units in the microfluidic chip 146, the number of specific structural components of the magnetic bead control layer middle layer 28 is also axial. Multiple sets of arc-shaped magnet channels are symmetrically arranged. The figure shows that it is preferable to set three sets of arc-shaped magnet channels with the same structure. The inner magnet 22 is disposed in an arc-shaped magnet channel, and the arc-shaped magnet channel can limit the movement of the inner magnet 22 in the arc-shaped magnet channel. Moreover, in each group of parallel detection units, the distance from the arc-shaped magnet channel to the center of the magnetic bead control layer middle layer 28 and the distance from the reaction chamber 8 to the center of the microfluidic chip middle layer 25 are consistent, and the arc-shaped magnet channel One end corresponds to the position of the reaction chamber 8 up and down. Figure 9 particularly shows an optimization strategy for rapid switching of reaction states through an enlarged view of the position of the spherical magnetic beads 23. The spherical magnetic beads 23 are arranged at the left and right ends of the arc-shaped magnet channel where the internal magnet 22 is located. The number of spherical magnetic beads 23 provided at each end of the internal magnet 22 is preferably two.

Regarding the strategy of rapid switching of the reaction state: through the movement of the internal magnet 22, the adsorption state of the magnetic beads in the reaction chamber 8 is switched, thereby completing different magnetic reactions such as adsorption and fixation of the magnetic beads or free mixing, and realizing the reaction. Real-time control and rapid switching of different reaction states of magnetic beads. Specifically, the magnetic bead control layer middle layer 28 is driven by the external magnet driving motor 152 to realize real-time control of the position of the internal magnet 22 and switching of reaction states. Specifically, the magnetic bead control layer 147 is placed close to the lower surface of the microfluidic chip 146, where the middle layer 28 of the magnetic bead control layer includes pin holes 7, light guide holes 21, internal magnets 22 and spherical magnetic beads 23; the internal magnet 22 is provided In the arc-shaped magnet channel, the distance from the arc-shaped magnet channel to the center of the magnetic bead control layer middle layer 28 is consistent with the distance from the reaction chamber 8 to the center of the microfluidic chip middle layer 25, and one end of the arc-shaped magnet channel Corresponds to the reaction chamber 8 of the microfluidic chip 146 up and down. The internal magnet 22 is located at one end of the reaction chamber 8 away from the microfluidic chip 146 in the arc-shaped magnet channel, which is called the distal end (as shown on the left in Figure 8 ). Correspondingly, when the internal magnet 22 is in the arc The end of the magnet channel located directly below the reaction chamber 8 of the microfluidic chip 146 is called the proximal end (as shown in the right picture in Figure 8 ). Combined with the partial enlarged view of the position of the spherical magnetic bead 23 in Figure 9, it can be seen that when the internal magnet 22 is at the proximal or distal end, the spherical magnetic bead 23 will exert a magnetic attraction on the internal magnet 22 to prevent it from being attached to the microfluidic chip 146 Movement during rotation affects the state of the magnetic beads.

During the reaction process, it is necessary to transfer the reagents through high-speed rotation or throw the waste liquid into the waste liquid chamber 17. In order to prevent the magnetic beads from being thrown into the next chamber in advance, the internal magnets 22 are required to adsorb and fix the magnetic beads. First, the external magnets 22 are required to adsorb and fix the magnetic beads. The magnet 154 is driven by the external magnet driving motor 152 and extends into the bottom of the magnetic bead control layer 147 in the horizontal direction, causing the magnetic bead control layer 147 to rotate counterclockwise. Under the influence of the magnetic force of 154, it automatically moves along the arc-shaped magnet channel to the proximal end (ie, below the reaction chamber 8), and efficiently adsorbs the magnetic beads, thus fixing the magnetic beads in the reagent. Secondly, during the reaction process, when the magnetic beads in the reagent and the sample need to be mixed, the external magnet 154, driven by the external magnet driving motor 152, extends into the bottom of the magnetic bead control layer 147 in the horizontal direction to control the magnetic beads. When the layer 147 rotates clockwise, the internal magnet 22 will automatically move to the far end (that is, away from the reaction chamber 8) along the arc-shaped magnet channel under the magnetic force of the external magnet 154, without adsorbing the magnetic beads, thereby realizing the freedom of the magnetic beads. Movement and mixing.

Figures 10 and 11 respectively show the left front side and the right rear perspective view of the internal structure of the device housing of the chemiluminescence measurement device of the present invention, mainly showing the positional relationship of each module in the internal structure of the device housing. It can be seen from the combination of Figure 10 and Figure 11 that the internal structure of the device shell of the chemiluminescence measurement device of the present invention includes a reagent module 45, an automatic pipetting module 44, an incubation chamber module 50, a chemiluminescence signal detection module 48, a centrifugal motor drive module 49 and an auxiliary module. , mainly fixed on the device bottom plate 47 of the device shell. Among them, the stepper motor driver 46, plunger pump 58, plunger pump fixing part 55, plunger pump solenoid valve 57 and plunger pump solenoid valve fixing part 56 in the automatic pipetting module 44 are all fixed on the device bottom plate 47 . It can also be seen in Figure 10 that an external magnet driving motor 152 is provided below the incubation chamber module 50. The external magnet driving motor 152 is fixed on the support column 79 and is used to drive the external magnet 154 (visible in conjunction with Figure 21) extending into the horizontal direction. Inside the incubation chamber module 50 and below the magnetic bead control layer 147, the internal magnet 22 is driven to rotate counterclockwise or clockwise in the arc-shaped magnet channel under the action of magnetic force, thereby achieving real-time control of the position of the internal magnet 22 Switching of reaction states with the magnetic bead control layer 147 .

The chemiluminescence measurement device of the present invention is also provided with an auxiliary module. The auxiliary module includes a main control board 52 and a main control board bracket 53. The main control board 52 is fixed on the main control board bracket 53. The main control board bracket 53 is fixed on the device shell. On the base plate 47, the main control board 52 is arranged in parallel with the rear side plate 39 of the device housing; the main control board 52 controls the operation of other modules and provides data interaction with the data interaction module.

Figures 10 and 11 also show the centrifugal motor drive module 49 and its centrifugal motor driver 54 respectively. A centrifugal motor driver partition 59 is provided on the front side of the centrifugal motor driver 54, wherein the centrifugal motor driver 54 and the centrifugal motor driver partition The plate 59 is also fixed on the device bottom plate 47, and the centrifugal motor driver partition 59 can avoid the potential impact that the heat of the centrifugal motor driver 54 may have on the reaction temperature. The centrifugal motor driver 54 is close to the rear side plate 39, and the heat generated is taken away in time through the heat sink 40 and its heat sink fan 41 provided on the rear side plate 39, thereby further ensuring temperature control while reducing the temperature interference to the device. accuracy.

It can be seen from Figure 10 and Figure 11 that the microfluidic chip module 51 is arranged in the middle of the incubation chamber module 50 . The drive shaft bearing 60 is a component in the X-axis direction of the automatic pipetting module 44 and can reduce the friction generated during the operation of the X-axis part of the automatic pipetting module 44.

Figure 12 shows a perspective view of the left front side of the reagent module 45 of the present invention, mainly showing the reagent box 61, the reagent box support plate 62 and the reagent box support frame 63 in the reagent module 45. It can be seen from the figure that the gun tip 68, the reagent test tube 66 and the washing liquid test tube 67 are all located in the corresponding holes on the reagent box 61. The gun tip 68, the reagent test tube 66 and the washing liquid test tube 67 on the reagent box 61 are arranged in parallel. There are multiple rows arranged, and the number of parallel arrangements is consistent with the number of parallel detection units on the microfluidic chip 146; a gun tip waste bin 65 is also provided on the test kit 61 to recover the waste after each sample addition. Gun tip; the reagent box 61 is fixed by the raised locking structure 64 on the reagent box support plate 62, and the reagent box support frame 63 is fixed on the device bottom plate 47 by screws.

The automatic pipetting module 44 includes automatic control in three directions: left and right, front and back, and up and down driven by three stepper motors: the X-axis stepper motor 73 controls the left and right movement in the horizontal direction (i.e., the X-axis); the Y-axis stepper controls The motor 99 controls the forward and backward movement in the horizontal direction (i.e., Y-axis); the Z-axis motor 117 controls the up-and-down movement in the vertical direction (Z-axis); that is, three sets of motors drive the automatic pipetting module 44 to achieve all-round movements of front and back, left and right, and up and down. Multi-angle pipetting movement.

Figures 13 and 14 respectively show a perspective view of the left rear side and the right front side of the automatic pipetting module 44 of the present invention, mainly showing the core components of the X-axis part of the automatic pipetting module 44. As can be seen in the figure, four support columns 79 are in a vertical "concave" shape and are fixed on the bottom plate 47 of the device. The automatic pipetting module 44 is fixed and supported above the reagent module 45 through the four support columns 79; automatic pipetting Two X-axis support plates are symmetrically arranged on the left and right sides of the module 44, namely, the X-axis left support plate 76 and the X-axis right support plate 87. These two X-axis support plates are symmetrically fixed on the support column 79 respectively. Specifically, the X-axis left support plate 76 and the X-axis right support plate 87 are in an "L" shape and are connected to the X-axis guide rails 81 at the left and right ends through screws. The two X-axis support plates are connected through a drive shaft 93.

In the X-axis part, the X-axis drag chain 75 is also fixed above the X-axis left support plate 76, and the X-axis stepper motor 73 is fixed on the end of the X-axis left support plate 76 close to the drive shaft 93 through the X-axis stepper motor holder 95. below. Further, the shaft of the X-axis stepper motor 73 and the drive shaft 93 are respectively fixed on the X-axis stepper motor synchronous wheel 94 and the drive shaft synchronous wheel 97. The two synchronous wheels are connected through the drive shaft belt 96, and through The X-axis stepper motor 73 drives the drive shaft 93 to rotate, thereby realizing movement in the Inside the X-axis bearing fixtures (X-axis right bearing fixture 92 and X-axis left bearing fixture 98), the drive shaft bearing 60 can reduce the movement between the two X-axis bearing fixtures and the drive shaft 93. Friction to achieve fast and smooth movement.

In the X-axis part, 2 driven wheels (X-axis left driven wheel 78 and X-axis right driven wheel 86) and 2 driving wheels (X-axis left driving wheel 69 and X-axis right driving wheel 91) are arranged symmetrically on the left and right. , and corresponding accessories; among them, the 2 driven wheels pass through 2 cantilever pins (X-axis left cantilever pin 84 and X-axis right cantilever pin 80) and 2 driven wheel fixing parts (X-axis left driven wheel fixing parts 77 and Axis right driven wheel fixing piece 85) is fastened; these two driven wheel fixing pieces are respectively fixed on the head of the X-axis left supporting plate 76 and the X-axis right supporting plate 87; the two driving wheels (X-axis left driving wheel 69 and The X-axis right driving wheel 91) is fixed on both ends of the drive shaft 93 by screws respectively. The drive shaft 93 is fixed by 2 bearing fixing parts (X-axis right bearing fixing part 92 and X-axis left bearing fixing part 98); 2 slave The moving wheel and the corresponding two driving wheels are connected using two X-axis belts 88 respectively. The two X-axis belts 88 are respectively pressed against the two X-axis tooth plates (X-axis left tooth plate) that are symmetrically arranged at the left and right positions. plate 74 and X-axis right tooth plate 90) and the corresponding two tooth plate fixing parts (X-axis left tooth plate fixing part 72 and X-axis right tooth plate fixing part 89), these two X-axis tooth plates and the corresponding The two tooth plate fixings are connected by screws; specifically, the X-axis left driven wheel fixing 77 and the X-axis right driven wheel fixing 85, the X-axis right bearing fixing 92 and the X-axis left bearing fixing 98 are all It is "L" shaped; further preferably, these two tooth plate fixing parts (X-axis left tooth plate fixing part 72 and X-axis right tooth plate fixing part 89) are in a vertical "concave" shape, with one end and 2 respectively. Two X-axis tooth plates (X-axis left tooth plate 74 and X-axis right tooth plate 90) are connected, and the other end is fixed on the Y-axis support plate 83 through screws, thereby achieving fastening of the Y-axis support plate 83.

In the X-axis part, an X-axis touch switch 71 is also provided at one end of the X-axis left support plate 76 near the drive shaft 93. The X-axis touch switch 71 is fixed on the X-axis touch switch fixing frame 70. The control switch fixing bracket 70 is further fixed on the X-axis left support plate 76, and is used to determine and control the initial position of the X-axis part through the X-axis touch switch 71, thereby achieving the purpose of limiting the position.

Figure 15 shows a side perspective view of the Y-axis part of the automatic pipetting module 44 of the present invention, mainly showing the core components of the Y-axis part. Combining Figure 13 and Figure 14, it can be seen that for the Y-axis part, firstly, the Y-axis support plate 83 is fixed on the X-axis guide rail slider 82 through screws; secondly, the stepper motor includes the X-axis stepper motor 73 and the Y-axis stepper motor 99 , the X-axis stepper motor 73 is fixed on the X-axis left support plate 76 through the ; The Y-axis driving wheel 101 is located on the central axis of the Y-axis stepper motor 99 and is fixed at the lower part of the Y-axis stepper motor bracket 100; further, the Y-axis stepper motor bracket 100 and the Y-axis driven wheel fixing member 107 are respectively Fixed on the left and right sides of the Y-axis support plate 83 with screws, the Y-axis driven wheel 106 is fastened through the Y-axis cantilever pin 108 and the Y-axis driven wheel fixing piece 107. Use Y-axis belt 102 to connect.

The Y-axis guide rail 116 is fixed on the Y-axis guide rail frame 105. The Y-axis guide rail 116 is also provided with a Y-axis guide rail slider 112. The Y-axis guide rail frame 105 is also fixed on the Y-axis support plate 83 through screws; the Y-axis drag chain 115 It is "U" shaped. One end of the "U" shaped opening of the Y-axis drag chain 115 is also provided with a Y-axis drag chain bracket 114. The Y-axis drag chain 115 is fixed on the Y-axis drag chain bottom plate 111 through screws. The Y-axis drag chain bottom plate 111 is then fixed on the rear side of the Y-axis support plate 83 through screws. The Y-axis belt 102 is pressed between the Y-axis tooth plate 104 and the convex structure behind the pipette bottom plate 103. The pipette bottom plate 103 is also provided with a pipette bottom plate spacer 113 on the upper part of the convex structure behind it. , the Y-axis tooth plate 104 and the pipette bottom plate 103 are connected through screws to realize the fastening of the Z-axis part of the automatic pipetting module 44.

In the Y-axis part, a Y-axis touch switch 109 is also provided. The Y-axis touch switch 109 is installed on the Y-axis touch switch holder 110. The Y-axis touch switch holder 110 is fixed on the Y-axis support plate 83 close to The upper end of one side of the Y-axis driven wheel 106 is used to determine and control the initial position of the Y-axis part through the Y-axis touch switch 109, thereby achieving the purpose of limiting the position.

Figure 16 shows a side perspective view of the Z-axis part of the automatic pipetting module 44 of the present invention, mainly showing the core components of the Z-axis part. As shown in Figure 16 combined with Figure 10 and Figure 15, in the Z-axis part, the Z-axis motor 117 is fixed on the top of the pipette base plate 103 through the Z-axis motor fixing piece 130, and the tail stop 125 of the pipette base plate is fixed on the pipette by screws. The tail of the pipette bottom plate 103; the Z-axis motor fixing piece 130 and the pipette bottom plate tail stop 125 are connected through two symmetrically arranged Z-axis guide shafts 119. The Z-axis guide shafts 119 are respectively connected to the Z-axis motor fixing piece through screws. 130 is fixed to the tail stop 125 of the pipette bottom plate, which plays a load-bearing and Z-axis direction guiding role; further, the two Z-axis guide shafts 119 each cooperate with a Z-axis guide shaft bearing 127, and the Z-axis guide shaft bearings 127 are symmetrical Embedded inside the pipette slider 121, the friction force generated during movement between the pipette slider 121 and the Z-axis guide shaft 119 can be reduced to achieve the purpose of fast and smooth movement. In the Z-axis part, the pipetting purpose is achieved by moving the pipette slider 121 up and down.

The top bracket 120 of the pipette slider, the pipette slider 121 and the bottom bracket 126 of the pipette slider are slidably arranged on the Z-axis guide shaft 119 from top to bottom, and are fixed to the Z-axis motor. The position between the pipette member 130 and the tail stop 125 of the pipette bottom plate; a spring 129 is placed between the pipette slider 121 and the pipette slider top bracket 120 to realize the elastic movement of the pipette slider 121; The top bracket 120 of the liquid container slider and the bottom bracket 126 of the pipette slider are fixed to the side connector 128 of the pipette slider through screws; the protruding "U" on the front side of the pipette slider 121 The shape structure and the pipette body 123 are fastened by pins. The hose connector 122 and the pipette 124 are respectively fixed on the upper and lower ends of the pipette body 123 through screws, that is, one end of the pipette body 123 is fixed to the pipette body 123 . The liquid pipe 124 is used to load and drop the gun tip; the other end of the pipette body 123 is fixed with a hose connector 122, and the hose connector 122 is connected to the plunger pump 58 through the air hose 164. The movement of the pipe causes the air pressure in the air hose 164 to increase or decrease, thereby controlling operations such as sampling and injection of samples or reagents through the air pressure, that is, controlling the pipetting action through the air pressure.

In the Z-axis part, a Z-axis touch switch 118 is also provided. The Z-axis touch switch 118 is fixed on the Z-axis motor fixture 130 through screws. The Z-axis touch switch 118 is used to determine and control the initial position of the Z-axis part. position to achieve the purpose of limiting.

Comprehensive observation of Figures 13 to 16 shows that in the automatic pipetting module 44, the three sets of motors in the three directions of the X-axis, Y-axis and Z-axis are: X-axis stepper motor 73, Y-axis stepper motor 99 and Z Axis motor 117, 3 sets of motors control the number of motor movement steps by converting electrical pulse signals into corresponding linear displacements, thereby achieving precise movement of the pipette in the three directions of the X-axis, Y-axis and Z-axis; on the X-axis The movement in the and Y-axis directions is realized by the X-axis guide rail 81 and the Y-axis guide rail 116 respectively. The movement in the Z-axis direction is realized by the Z-axis motor 117. The Z-axis motor 117 is a screw motor; for the X-axis, Y-axis and Z-axis The 3 sets of motors in the 3 directions of the axis are also equipped with 3 corresponding touch switches, namely X-axis touch switch 71, Y-axis touch switch 109 and Z-axis touch switch 118, which are used to determine the zero point position of movement, which is flexible Conveniently realize operations such as loading and dropping the gun tip, sampling and injection of samples.

Figures 17, 18 and 19 respectively show a side perspective view of the incubation chamber module 50 of the present invention and a partial structural view of the heating upper cover and the sealing lower cover. The incubation chamber module 50 (see Figure 10) uses an aluminum block heating method to control the temperature of the reaction. Figure 17 shows that the incubation chamber module 50 includes a slide motor 133, a heating upper cover 134 and a sealing lower cover 135; among them, the slide motor 133 is fixed on the device bottom plate 47 (see Figure 10) through the slide motor support frame 132. The heating upper cover 134 and the slide motor 133 are connected through the heating upper cover connector 131. The slide motor 133 is controlled by the heating upper cover connector 131. The upper heating cover 134 moves vertically in the Y-axis direction, and the height of the upper heating cover 134 can be adjusted within a range of 50 mm. In this way, before placing the microfluidic chip module 51, the upper heating cover 134 is in a lifted state and will not It affects the placement action of the microfluidic chip module 51. After the placement of the microfluidic chip module 51 is completed, the heating upper cover 134 falls and is tightly fastened to the sealing lower cover 135 to form an incubation chamber.

As shown in Figure 18, there is a layer of heating film 137 closely attached to the heating aluminum block 136. The heating film 137 corresponds to the position of the heating aluminum block 136; the heating aluminum block 136 is fixed on the inside of the heating upper cover 134 through screws. At the same time, the heating upper cover 134 moves downward until it is wrapped by the sealing lower cover 135, thereby forming a sealed, dual-function incubation chamber. On the one hand, the incubation chamber can provide a closed temperature-controlled environment to achieve accurate temperature control of the microfluidic chip module 51 and avoid interference and influence caused by ambient temperature fluctuations to the greatest extent; on the other hand, the incubation chamber can also provide A light-shielding environment can minimize the interference and impact of stray light in the environment on chemiluminescence signal collection, improve the sensitivity of light signal collection, and achieve accurate signal collection and analysis. In addition, in the incubation chamber module 50, circular through holes are provided in the middle of the heating upper cover 134, the heating film 137, the heating aluminum block 136 and the sealing lower cover 135, and between the heating upper cover 134, the heating film 137 and the heating aluminum block 136 The sealing lower cover 135 is fixed on the centrifugal motor bracket 151 (see Figure 21) through screws; a capsule-shaped through structure 163 is provided at the corresponding position of the heating upper cover 134, the heating film 137 and the heating aluminum block 136, so that The light leakage area of the microfluidic chip can be minimized. The upper heating cover 134, the heating film 137 and the heating aluminum block 136 are all circular in shape. The heating film is customized and can be tightly adhered to the upper surface of the heating aluminum block 136 using double-sided tape.

Figure 19 shows that the surface of the sealing lower cover 135 is provided with a light pipe channel 138, and the side of the sealing lower cover 135 is provided with an external magnet channel 139. The bottom of the sealing lower cover 135 is fixedly provided with a positioning optical coupler 140, and the positioning optical coupler 140 is provided with a light channel. , the sealing lower cover 135 is also provided with a positioning optocoupler wiring hole 141 on the side away from the external magnet channel 139 .

Figure 20 shows a front view of the chemiluminescence signal detection module 48 of the present invention. The chemiluminescence signal detection module 48 collects the chemiluminescence signal through the photomultiplier tube 143, and the sensitivity is relatively high; in the chemiluminescence signal detection module 48, it is also provided There is a light guide 142. The light guide 142 is cylindrical and is composed of a jacket (PVC) wrapping an optical fiber (PMMA). The chemiluminescence signal detection module 48 is provided with the light guide 142, which not only solves the problem that the photomultiplier tube 143 cannot be placed directly under the microfluidic chip module 51 to detect the chemiluminescence signal, but also can detect the weak chemical generated by the reaction. The luminescent signal is quickly collected and transmitted, which avoids the loss of weak luminescent signal in the internal space of the instrument and improves the accuracy of detection. One end of the light guide tube 142 is fastened to the photomultiplier tube 143 through threads. The chemiluminescence signal detection module 48 is fixedly installed on the device bottom plate 47 through the photomultiplier tube 143 and is located below the sealing lower cover 135 in the incubation chamber module 50. The other end of the light guide 142 extends into the lower side of the alignment microfluidic chip module 51 through the light guide channel 138 (see Figure 19) of the sealed lower cover 135, and is aligned with the detection cavity 14 of the microfluidic chip 146. To detect the chemiluminescence signal generated by the microfluidic chip 146. During the detection process, when the magnetic beads, pre-excitation liquid and excitation liquid enter the detection chamber 14 to react and emit light, the weak light signal generated will be transmitted to the photomultiplier tube 143 through the light guide 142. The photomultiplier tube 143 amplifies and converts weak optical signals into electrical signals, thereby realizing the collection and detection of chemiluminescence signals, and improving the sensitivity of chemiluminescence signal collection.

Figure 21 shows a perspective view of the internal structure of the centrifugal motor drive module 49 of the present invention after removing the sealing lower cover 135. It is located inside the sealing lower cover 135. The structure of the centrifugal motor drive module 49 (see Figure 10) can be seen from Figure 21: centrifugal motor The bracket 151 is fixed on the bottom plate 47 of the device. The centrifugal motor 150 is fixed on the centrifugal motor bracket 151 through screws. The flange base 149 is nested above the centrifugal motor bracket 151 and is located on the central axis of the centrifugal motor 150. Driven by the centrifugal motor 150 , driving the components fixed on the flange base 149 to rotate and centrifuge around the central axis of the centrifugal motor 150; specifically, the microfluidic chip module 51 (including the microfluidic chip 146 and the magnetic bead control layer 147) passes through the butterfly The screws 144 and the chip pressing member 145 are fixed on the flange base 149 above the centrifugal motor 150, and the butterfly screws 144, the chip pressing member 145, the microfluidic chip 146, the magnetic bead control layer 147 and the flange base 149 are from They are located above the central axis of the centrifugal motor 150 from top to bottom, with the central axis of the centrifugal motor 150 as the center. The centrifugal motor 150 is used to drive the microfluidic chip module 51 and other components to perform rotation and centrifugation.

Further, the positioning optical coupler 140 is fixed on the bottom of the sealing lower cover 135, and the positioning optical coupler 140 is provided with a light channel. The upper part of the centrifugal motor 150 is also provided with a grating sheet 148. The grating sheet 148 is fixed on the side of the flange base 149 through screws, and the grating sheet 148 is provided with a slit with a width of 0.1 mm-0.5 mm. The slit is related to the positioning. The optical channel of the optocoupler 140 (see Figure 19) corresponds to the positioning of the microfluidic chip 146.

Located outside the sealed lower cover 135, the structure of the centrifugal motor drive module 49 (see Figure 10) can be seen from Figure 21: the external magnet 154 is fixed at one end of the external magnet fixing part 153, and the external magnet fixing part 153 is connected to the external magnet driving motor 152. Connection, the external magnet driving motor 152 is fixed on the support column 79 through pins and screws, and the external magnet controlling motor 152 is used to drive the external magnet 154 and control the movement state of the internal magnet 22 during the reaction process. In addition, when the centrifugal motor 150 is in a non-centrifugal state, the external magnet 154 can extend into the interior of the sealing lower cover 135 through the external magnet channel 139 provided on the side of the sealing lower cover 135, and the internal magnet 22 in the microfluidic chip module 51 Below, the positions of the external magnet 154 and the internal magnet 22 correspond up and down. Driven by the external magnet driving motor 152, the external magnet 154 performs magnetic control on the corresponding internal magnet 22, thereby controlling the switching of the reaction state of the magnetic bead control layer 147. .

Figure 22 shows a three-dimensional structural view of the core components of the external magnet drive motor 152 of the present invention, which mainly includes a DC motor touch switch fixing part 155, a DC motor touch switch 156, a guide rail frame 157, a guide rail bearing 158, and a DC motor slider 159. , guide rail 160, DC motor 161 and DC motor bracket 162, the external magnet control motor 152 is fixed on the support column 79 at a suitable height through a connecting piece, making the layout of the internal parts of the instrument more compact. Among them, the guide rail bearing 158 is symmetrically embedded and installed inside the DC motor slider 159, which can reduce the friction between the DC motor slider 159 and the guide rail 160 to achieve the purpose of fast and smooth movement. The DC motor slider 159 passes through the connecting piece. The external magnet 154 is fixed, and the external magnet drive motor 152 is also provided with a DC motor touch switch 156. The DC motor touch switch 156 is fixed on the DC motor touch switch fixture 155 and is used to determine and control the start of the external magnet drive motor 152. and stop.

The chemiluminescence measurement device of the present invention creatively integrates a microfluidic chip module with multiple parallel detection units, an automatic pipetting module that drives three sets of motors to move in three directions, and a centrifugal motor and an external magnet drive motor. Centrifugal motor drive module and other unique structures. In addition, the present invention also provides a controllable closed and light-shielding environment for the microfluidic chip module through the incubation chamber and its up and down movement driven by the slide motor. It can not only effectively control and maintain the temperature of the device, but also effectively avoid all troubles. The interference and influence of astigmatism on chemiluminescence signal collection. The final innovative and optimized chemiluminescence measurement device not only has the advantages of small size, low cost, high throughput, simple structure, convenient operation, high sensitivity and accurate results, but also can be fully automated and multiple jointly tested flexibly into the micro Samples to be tested in the fluidic chip module.

Claims (9)

1. A chemiluminescent assay device comprising a device housing and a device disposed within the device housing:

the micro-fluidic chip module (51) comprises a micro-fluidic chip (146) and a magnetic bead control layer (147), wherein the micro-fluidic chip is of a three-layer structure, multiple groups of parallel detection units with the same structure are arranged in an axisymmetric manner, the three-layer structure of the micro-fluidic chip (146) is respectively a micro-fluidic chip cover plate (24), a micro-fluidic chip middle layer (25) and a micro-fluidic chip bottom plate (26), the three-layer structure of the magnetic bead control layer (147) is respectively a magnetic bead control layer cover plate (27), a magnetic bead control layer middle layer (28) and a magnetic bead control layer bottom plate (29), and each group of parallel detection units of the micro-fluidic chip middle layer (25) sequentially comprises a reaction cavity (8), a buffer cavity (12), a detection cavity (14) and a waste liquid cavity (17); the magnetic bead control layer (147) is tightly attached to the lower surface of the microfluidic chip (146), and a circular arc-shaped magnet channel, an internal magnet (22) in the circular arc-shaped magnet channel and spherical magnetic beads (23) at the left end and the right end of the circular arc-shaped magnet channel are arranged on the middle layer (28) of the magnetic bead control layer;

The reagent module (45) comprises gun tips (68), reagent test tubes (66) and washing liquid test tubes (67), and the reagent module is arranged in a plurality of rows which are arranged in parallel, and the number of the parallel rows is consistent with the number of parallel detection units on the microfluidic chip (146);

the automatic pipetting module (44) is driven by 3 sets of motors respectively to control the pipetting movement of front and back, left and right, up and down in all directions and the movement in the X-axis and Y-axis directions is realized by an X-axis guide rail (81) and a Y-axis guide rail (116) respectively: the X-axis direction is provided with 2X-axis guide rails (81) and 2 corresponding X-axis guide rail sliding blocks (82), 2X-axis support plates are symmetrically arranged and connected with the X-axis guide rails (81) at the left end and the right end, the 2X-axis support plates are connected through driving shafts (93), shafts of X-axis stepping motors (73) and the driving shafts (93) are respectively fixed on X-axis stepping motor synchronous wheels (94) and driving shaft synchronous wheels (97), the X-axis stepping motor synchronous wheels (94) are connected with the driving shaft synchronous wheels (97) through driving shaft belts (96), and the X-axis stepping motors (73) drive the driving shafts (93) to rotate, so that left-right movement of the X-axis stepping motors (73) in the horizontal direction is realized; the Y-axis guide rail (116) is fixed on a Y-axis guide rail frame (105), a Y-axis guide rail sliding block (112) is further arranged on the Y-axis guide rail (116), the Y-axis guide rail frame (105) is fixed on a Y-axis supporting plate (83), the Y-axis supporting plate (83) is fixed on an X-axis guide rail sliding block (82), 2X-axis belts (88) are respectively pressed between 2X-axis toothed plates which are symmetrically arranged and 2 toothed plate fixing pieces which are vertical concave, one end of each of the 2 toothed plate fixing pieces is respectively connected with the 2X-axis toothed plates, and the other end of each of the 2 toothed plate fixing pieces is fixed on the Y-axis supporting plate (83), so that the Y-axis connection is realized and the horizontal direction front-back movement which is driven and controlled by a Y-axis stepping motor (99) is realized; the vertical direction of the Z-axis motor (117) is controlled to move up and down, the Z-axis motor (117) is a screw motor, a pipette (124) is fixed at one end of a pipette body (123) in the Z-axis direction, a hose connector (122) is fixed at the other end of the pipette body (123), the hose connector (122) is connected with a plunger pump (58) through a gas path hose (164), the gas pressure in the gas path hose (164) is controlled through the movement of the plunger pump (58), and then the pipetting action of the pipette body (123) is controlled through the gas pressure; the automatic pipetting module (44) is fixed and supported above the reagent module (45) by a support column (79);

The incubation bin module (50) adopts an aluminum block heating mode, comprises a sliding table motor (133) and a heating aluminum block (136), and constructs a closed and shading incubation reaction bin for the microfluidic chip (146) by arranging a heating upper cover (134) and a sealing lower cover (135); the heating aluminum block (136) is fixed on the inner side of the heating upper cover (134), a heating film (137) is adhered to the upper surface of the heating aluminum block (136), the heating film (137) corresponds to the center position of the heating aluminum block (136), and a capsule-shaped through structure (163) is arranged at the corresponding positions of the heating upper cover (134), the heating film (137) and the heating aluminum block (136);

a centrifugal motor driving module (49) comprising a centrifugal motor (150) positioned inside the sealed lower cover (135), a centrifugal motor bracket (151), a flange base (149), and an external magnet (154), an external magnet fixing member (153) and an external magnet driving motor (152) positioned outside the sealed lower cover (135); the external magnet driving motor (152) is fixed on the supporting column (79) and is used for driving the external magnet (154) to magnetically control the corresponding internal magnet (22); the centrifugal motor (150) is one of a servo motor, a stepping motor, a DC brushless motor or other motors to provide a rotational speed in the range of 1000 rpm-12000 rpm;

One end of the circular arc-shaped magnet channel corresponds to the position of the reaction cavity (8) up and down, the inner magnet (22) moves to one end of the micro-fluidic chip (146) right below the reaction cavity (8) in the circular arc-shaped magnet channel, which is called a near end, the inner magnet (22) moves to one end of the reaction cavity (8) far away from the micro-fluidic chip (146) in the circular arc-shaped magnet channel, which is called a far end, in a non-centrifugal state, the outer magnet (154) stretches into the lower part of the inner magnet (22) in the sealing lower cover (135) through an outer magnet channel (139) arranged on the side surface of the sealing lower cover (135), and the magnetic force controls the inner magnet (22) to move in the circular arc-shaped magnet channel, so that the switching of the reaction state of the magnetic bead control layer (147) is controlled.

2. The chemiluminescent assay of claim 1 wherein: the three-layer structures of the microfluidic chip (146) are respectively provided with a bonding positioning hole (6) and/or a pin hole (7) with corresponding positions, and the three-layer structures are fixed through the bonding positioning holes (6) and/or the pin holes (7);

The reaction cavity (8) is connected with the detection cavity (14) through a first capillary valve (19), the buffer cavity (12) is connected with the detection cavity (14) through a buffer cavity liquid channel (13), the detection cavity (14) is connected with the waste liquid cavity (17) through a second capillary valve (16) with a circular blocking valve (15), and magnetic beads, samples or reagents sequentially complete all steps of chemiluminescent reaction in all the cavities;

the reaction cavity (8) is provided with a reaction cavity air hole channel (18), the buffer cavity (12) is provided with a buffer cavity air hole channel (10), and the detection cavity (14) is provided with a detection cavity air hole channel (20);

the reaction chamber (8) is provided with a reaction chamber sample adding channel (9), the buffer chamber (12) is provided with a buffer chamber sample adding channel (11), the buffer chamber sample adding channel (11) is used for adding excitation liquid into the buffer chamber (12), and the excitation liquid is thrown into the detection chamber (14) through a buffer chamber liquid channel (13) under the centrifugal force action of the centrifugal motor (150) to perform luminous reaction.

3. The chemiluminescent assay of claim 1 wherein: the microfluidic chip module (51) is fixed on the flange base (149) above the centrifugal motor (150), the flange base (149) is nested on the central shaft of the centrifugal motor (150) above the centrifugal motor bracket (151), and under the driving of the centrifugal motor (150), the component fixed on the flange base (149) is driven to rotate and centrifuge around the central shaft of the centrifugal motor (150).

4. The chemiluminescent assay of claim 1 wherein: the sliding table motor (133) controls the heating upper cover (134) to move up and down in the Z-axis direction, and the height of the heating upper cover (134) is adjusted.

5. The chemiluminescent assay of claim 1 wherein: the middle layer (28) of the magnetic bead control layer is also provided with a pin hole (7) and a light guide hole (21); under the drive of an external magnet driving motor (152), the magnetic bead control layer (147) rotates anticlockwise, the internal magnet (22) moves to the near end right below the reaction cavity (8) in the circular arc-shaped magnet channel, and the magnetic beads in the reaction cavity (8) are fixed; under the drive of an external magnet driving motor (152), the magnetic bead control layer (147) rotates clockwise, the internal magnet (22) moves to the far end far away from the reaction cavity (8) in the circular arc-shaped magnet channel, and the magnetic attraction fixation of the magnetic beads in the reaction cavity (8) is released, so that the magnetic beads move freely in the reaction cavity (8); the spherical magnetic beads (23) generate magnetic attraction to the internal magnets (22), and the number of the spherical magnetic beads is 2.

6. The chemiluminescent assay according to any one of claims 1-5 wherein: the device shell is made of a metal material with heat conducting performance, a radiating fin (40) is arranged on a rear side plate (39) of the device shell, and a radiating fin fan (41) is fixedly arranged in the middle of the radiating fin (40);

A data interaction module and a power module are further arranged above the device shell, and the data interaction module provides a man-machine interaction interface through a touch screen (30); the power module is matched with the power adapter to provide stable output power.

7. The chemiluminescent assay according to any one of claims 1-5 wherein: in the incubation bin module (50), the sliding table motor (133) is fixed on a device bottom plate (47) of the device shell through a sliding table motor support frame (132), and the heating aluminum block controls the temperature of the incubation bin module through a PID algorithm;

the heating aluminum block (136) moves to the upper side of the micro-fluidic chip (146) along with the heating upper cover (134), heats the micro-fluidic chip (146), and controls the temperature of the incubation bin module (50).

8. The chemiluminescent assay according to any one of claims 1-5 wherein: the device is also provided with a chemiluminescent signal detection module (48) which comprises a photomultiplier (143) and a light pipe (142), wherein one end of the light pipe (142) is fastened with a window of the photomultiplier (143) and is fixed on a device bottom plate (47) of the device shell; the other end of the light pipe (142) extends below the microfluidic chip module (51) through a light pipe channel (138) arranged on the surface of the sealing lower cover (135), is aligned to the detection cavity (14) of the microfluidic chip (146) and is used for detecting a chemiluminescent signal generated by the microfluidic chip (146);

A grating sheet (148) is arranged at the upper part of the centrifugal motor (150), a slit is arranged on the grating sheet (148), and the grating sheet (148) is fixed on the side surface of the flange base (149); a positioning optocoupler (140) is arranged at the bottom of the sealing lower cover (135), and the positioning optocoupler (140) is provided with an optical channel; the slit corresponds to the optical channel for positioning the microfluidic chip (146).

9. Use of a chemiluminescent assay device according to any one of claims 1-8 in chemiluminescent immunoassay.