CN103675306A - Automatic analysis instrument and system for microfluidic immunodetection chip - Google Patents

Abstract

The invention discloses an automatic analysis instrument and system for a microfluidic immune detection chip. The automatic analysis instrument includes an instrument frame, a mechanical movement module fixed on the instrument frame, a liquid pipetting module and a detection module; The mechanical movement module includes a three-dimensional mechanical arm and a metal working tube fixed on the three-dimensional mechanical arm, the movement of the three-dimensional mechanical arm drives the metal working tube to move for the assembly and removal of the suction head; the liquid transfer The extraction module is connected to the metal working tube for liquid suction and injection; the detection module is used for reading and transmission of chemiluminescence signals on the microfluidic immune detection chip. The instrument is connected to a computer to form a system, which fully realizes the organic integration of sample pretreatment, target protein separation and purification, sample and reagent injection and aspiration, chip cleaning, signal acquisition and analysis processing, making microfluidic technology automatic, miniaturized and more accurate and easier.

Description

Automated analysis instrument and system for microfluidic immunoassay chip

technical field

The invention relates to the technical field of microfluidic chip analysis systems, in particular to an automatic analysis instrument and system for microfluidic immune detection chips.

Background technique

Microfluidic immunoassay method is a newly developed technology in recent years. This method is mainly based on analytical chemistry, combined with the achievements of related disciplines such as biochemistry, physical chemistry and immunology, and manipulates nanoliter to Fluid in picoliter volume, for immunoreaction detection, has the advantages of small volume, large specific surface area, short reaction time, fast analysis speed, less reagent and sample consumption, and simultaneous detection of multiple samples and multiple indicators. It has important practical value for the discovery of disease markers and early diagnosis of diseases in clinical disease proteomics research. Conventional immunoassays require a relatively long analysis time, the liquid handling process is also cumbersome, the throughput is relatively small (only one or a few samples can be detected at a time), and more antibody reagents are required. The microfluidic analysis chip can effectively overcome these shortcomings. For example, when analyzing samples with very small sample volumes, microfluidic technology shows great advantages. Usually, experiments that require a few milliliters of sample volume only need a few microliters of sample volume after using microfluidic technology, which is greatly improved. Consumption of samples and reagents is saved. Moreover, the combination of microfluidic analysis technology and immune analysis can also overcome other shortcomings of traditional immune analysis to a certain extent, so it has attracted extensive attention of researchers in recent years. The application of microfluidic technology in immunoassay will greatly promote the improvement of immunoassay technology, and has broad application prospects and important application value.

However, in the current microfluidic immunoassay method, in addition to the steps that can be completed on the chip, other operations (such as sample pretreatment, introduction, cleaning, and signal readout detection) mostly need to be done manually, and the steps are cumbersome. The workload is increased, and detection errors are prone to occur, so it cannot be conveniently applied to real-time field detection, and it is not convenient for the wide application and promotion of microfluidic detection methods. Moreover, it cannot fully reflect the advantages of microfluidics, which is essentially high-throughput detection in this regard. Therefore, it is necessary to develop a fully automatic microfluidic detection and analysis instrument that fully realizes the organic integration of sample pretreatment, target protein separation and purification, sample and reagent injection and aspiration, chip cleaning, signal acquisition and analysis processing, making microfluidic Control technology automation, miniaturization, more accurate and simpler.

Contents of the invention

The purpose of the present invention is to provide an automatic analysis instrument and system for microfluidic immunoassay chip, which fully realizes sample pretreatment, target protein separation and purification, sample and reagent injection and aspiration, chip cleaning, signal acquisition and The organic integration of analysis and processing makes microfluidic technology automatic, miniaturized, more accurate and easier.

For realizing the purpose of the present invention, provide following technical scheme:

An automatic analysis instrument for a microfluidic immune detection chip, comprising an instrument frame and a mechanical movement module fixed on the instrument frame, a liquid pipetting module and a detection module; the mechanical movement module includes a three-dimensional mechanical arm and a fixed The metal working tube on the three-dimensional mechanical arm, the movement of the three-dimensional mechanical arm drives the metal working tube to move for the assembly and removal of the suction head; the liquid pipetting module communicates with the metal working tube for liquid absorption and injection; the detection module is used for reading and transmitting chemiluminescent signals on the microfluidic immunodetection chip.

Preferably, the mechanical movement module also includes a working platform, on which a chip slot, a reagent slot and a tip slot are arranged, the chip slot is used to place a microfluidic immunoassay chip, and the reagent slot is used to place As for the reagent bottle, the tip groove is used to place the tip for the assembly of the metal working tube.

Preferably, the chip tank, the reagent tank and the tip tank are arranged side by side in sequence, and a waste liquid tank is arranged behind the tip tank.

Preferably, a suction tip slot is provided on the edge of the waste liquid tank for the suction tip on the metal working tube to be stuck.

Preferably, the liquid removal module includes a stepping motor, a syringe pump controlled by the stepping motor, and a catheter, and the syringe pump communicates with the metal working tube through a catheter.

Preferably, the detection module is a CCD camera, and the CCD camera is fixed on the three-dimensional manipulator and moves with the movement of the three-dimensional manipulator.

Preferably, the three-dimensional mechanical arm is composed of three sets of servo motors in the directions of X, Y and Z axes, lead screws and guide rails.

Preferably, the instrument frame is a light-tight structure.

Preferably, the instrument frame is mainly composed of a metal frame and a metal plate.

Preferably, the CCD camera is connected to a computer, and the computer receives and processes the chemiluminescent signal transmitted by the CCD camera.

An automatic analysis system for a microfluidic immunoassay chip, comprising the above-mentioned automatic analysis instrument and a computer connected thereto.

The beneficial effects of the present invention are: the automatic analysis instrument for the microfluidic immune detection chip of the present invention includes an instrument frame and a mechanical movement module fixed on the instrument frame, a liquid pipetting module and a detection module; The motion module includes a three-dimensional mechanical arm and a metal working tube fixed on the three-dimensional mechanical arm. The movement of the three-dimensional mechanical arm drives the metal working tube to move for the assembly and removal of the suction head; the liquid pipetting module The metal working tube is connected to be used for drawing and injecting the liquid; the detection module is used for reading and transmitting the chemiluminescent signal on the microfluidic immune detection chip. The above-mentioned automatic analysis instrument is connected to a computer to form an automatic analysis system, which can align the microfluidic pipeline inlet, fix the position of the reagent, absorb and dispense the liquid, and read out and analyze the chemiluminescent signal for the microfluidic immune detection chip. Functions, fully realize the organic integration of sample pretreatment, target protein separation and purification, sample and reagent injection and aspiration, chip cleaning, signal acquisition and analysis processing, making microfluidic technology automatic, miniaturized, more accurate and easier. Using the automated analysis system can realize high-throughput detection with simple operation and low workload.

Description of drawings

Fig. 1 is the top view of the main structure of the automatic analysis instrument for microfluidic immune detection chip of the present invention, wherein, 1 represents a three-dimensional mechanical arm, 2 represents a metal working tube, 3 represents a working platform, 4 represents a CCD camera, and 31 represents a chip groove , 32 indicates the reagent tank, 33 indicates the tip tank, 34 indicates the waste liquid tank, and 35 indicates the tip card slot.

2 is a side view of the main structure of the automatic analysis instrument for microfluidic immune detection chip of the present invention, wherein, 1 represents a three-dimensional mechanical arm, 2 represents a metal working tube, 3 represents a working platform, 4 represents a CCD camera, and 35 represents a suction head slot.

Fig. 3 is a photo of the results read by the automatic analysis system for the microfluidic immunoassay chip of the present invention, wherein the white squares represent chemiluminescent signals, and brighter white squares represent stronger chemiluminescent signals.

Detailed ways

Embodiments of the present invention will be described in detail below through specific embodiments. Those skilled in the art will understand that the following embodiments are only preferred embodiments of the present invention for a better understanding of the present invention, and thus should not be considered as limiting the scope of the present invention. For those skilled in the art, the present invention can have various modifications and changes, and within the spirit and principles of the present invention, any modifications, equivalent replacements or improvements, etc., should be included in the protection scope of the present invention within.

Please refer to Figure 1 and Figure 2, the automatic analysis system for microfluidic immunoassay chip is mainly composed of instrument frame, mechanical movement module, liquid pipetting module, detection module and computer (not shown in the figure). Among them, the instrument frame is used to fix the position of each component of the instrument; the mechanical movement module is used for the three-dimensional movement of the position of the metal working tube, as well as the assembly and removal of the suction head on the metal working tube; the liquid pipetting module is used for the absorption of trace liquids and injection; the detection module is used to read the chemiluminescent signal on the microfluidic chip, and transmit the image information to the computer. The effective integration of mechanical movement module, liquid pipetting module, detection module and computer is realized. This system meets the requirements of immunoassay on microfluidic chip. It is a new type of fully automatic microfluidic chip analysis system.

Each component of the automated analysis system will be described in detail below with reference to FIG. 1 and FIG. 2 .

The instrument frame is mainly composed of metal frame and metal plate. Since the chemiluminescent signal detection of the microfluidic chip needs to be protected from light, the frame of the instrument is designed as a light-tight structure, and an openable door structure (not shown in the figure) is designed on the front side of the instrument for the removal of chips and reagents. The mechanical movement module of the main structure of the system is fixed on the instrument frame.

The mechanical movement module mainly includes a three-dimensional mechanical arm 1 , a metal working tube 2 and a working platform 3 . The three-dimensional robotic arm 1 is composed of three sets of servo motors in the directions of X, Y and Z axes, lead screws and guide rails (not shown in the figure). The bottom structure of the metal working tube 2 is designed according to the structure of the top of the suction head, and can be closely matched with the suction head. The top of the metal working tube 2 is sealed and connected with the syringe pump and the catheter to meet the airtightness requirements of the liquid pipeline. A chip groove 31, a reagent groove 32, a tip groove 33 and a waste liquid groove 34 are arranged on the working platform 3, the chip groove 31 is used to place a microfluidic immune detection chip, and the reagent groove 32 is used to place a reagent bottle. The suction tip tank 33 is used to place suction tips for metal working tube assembly, and the waste liquid tank 34 is used to contain system waste liquid. The edge of the waste liquid tank 34 is provided with a suction tip clamping groove 35, which is designed according to the diameter of the tip of the suction tip for the clamping of the suction tip on the metal working tube 2.

The liquid pipetting module is composed of a syringe pump controlled by a stepping motor and a catheter. The syringe pump communicates with the metal working tube 2 through the catheter, and can accurately absorb and inject 3-100 μL of liquid.

The detection module is a CCD (Charge-coupled Device) camera 4, also known as a CCD image sensor, which is a semiconductor device that can convert optical images into digital signals and complete the detection of chemiluminescent signals on the microfluidic chip. Image and transfer to computer. In addition, any camera used to read the chemiluminescent signal on the microfluidic chip and transmit the image information to the computer can be used as the detection module of the present invention.

The working principle and process of the automatic analysis system for the microfluidic immune detection chip are as follows: the microfluidic chip is placed in the chip slot 31, the reagent used in the test is placed in the reagent slot 32, and the suction tip is placed in the tip slot 33 In standby mode, start the instrument, execute the command after writing the relevant program. Under the control of the program, after the three-dimensional robotic arm 1 moves the metal working tube 2 to the position directly above the suction head, the Z axis moves downward to closely connect with the suction head, and the metal working tube 2 is moved to the reagent by the movement of the three-dimensional mechanical arm 1 Directly above the bottle, the Z axis moves downward, immerse the suction head on the bottom end of the metal working tube 2 below the liquid level of the reagent bottle, suck the quantitative liquid through the syringe pump, and move it directly above the injection port of the microfluidic chip, The Z-axis moves downward to realize the close connection between the suction head and the microfluidic chip inlet, and injects quantitative liquid into the microfluidic chip through the syringe pump. This process is a liquid suction and injection action. Through the orderly By repeating, all the liquid manipulation work of microfluidic chip immunoassay can be realized. The microfluidic chip is designed with a waste liquid pool, which can effectively collect and store waste liquid. After the immunoassay reaction, the three-dimensional robotic arm 1 moves the CCD camera 4 directly above the reaction area of the chip, and the CCD camera 4 converts the chemiluminescent signal on the chip into an electrical signal, which is then transmitted to the computer for data processing and analysis.

After testing, the system has high three-axis movement precision, and can realize the precise docking of the suction head and the microfluidic chip inlet, as well as the docking of the metal working tube 2 and the suction head, with a movement accuracy of 0.01mm; the system With high liquid control precision, it can realize the precise suction and injection of 3μL-100μL liquid, and the error of pipetting accuracy is ±5%. High-efficiency reading, the exposure time can reach more than 50min; the system has a high automatic control ability, after programming, all the immunoassay tests can be operated with one key; the system is simple and compact in structure, small in size, Ease of use.

The system also has the following features: use the suction head to complete the liquid transfer action, avoid mutual contamination, and save cleaning steps; use high-precision servo motors to effectively control the three-dimensional position of the metal working rod; Chemiluminescent signal on the flow control chip; reasonable design, realizes the optimization and integration of mechanical movement module, liquid pipetting module and detection module, realizes the integration of the system, small size, easy to detect and operate; fully automatic design, easy to operate , realizes the automatic operation of the microfluidic chip immunoassay test, and only needs one key to complete the entire operation process.

The automatic analysis system for the microfluidic immune detection chip of the present invention is used to detect the chemiluminescent signal after the reaction of the microfluidic immune detection chip. Specifically, the alpha-fetoprotein (AFP)-coated antibody was pre-immobilized on the 1 and 2 directions of the microfluidic immunoassay chip, and the blocking solution (small bovine serum albumin, etc.), AFP solution, cleaning solution (phosphate buffer, etc.), AFP detection antibody, cleaning solution and chemiluminescent solution (hydrogen peroxide and luminol, etc.), of which A, B, C and D tubes The concentration of the added AFP solution was 200ng/mL, 100ng/mL, 50ng/mL and 25ng/mL in sequence. The photo of the reading result (Figure 3) shows: the higher the concentration of AFP solution, the stronger the chemiluminescence signal, and the brighter the white square.

The applicant states that the present invention illustrates the detailed features and detailed methods of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned detailed features and detailed methods, that is, it does not mean that the present invention must rely on the above-mentioned detailed features and detailed methods. implement. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of selected components of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the scope of protection and disclosure of the present invention.

Claims (10)

1. for an automated analysis instrument for micro-fluidic immune detection chip, it is characterized in that, comprise apparatus frame and the mechanical motion module, the liquid that are fixed on described apparatus frame pipette module and detection module; Described mechanical motion module comprises three-dimensional machinery arm and is fixed on the working metal pipe on described three-dimensional machinery arm, and assembling and card that the motion of described three-dimensional machinery arm drives described working metal pipe to move for suction nozzle fall; Described liquid pipettes module and is communicated with described working metal pipe for absorption and the injection of liquid; Described detection module reads and transmits for chemiluminescence signal on micro-fluidic immune detection chip.

2. automated analysis instrument according to claim 1, it is characterized in that, described mechanical motion module also comprises workbench, chip groove, reagent trough and suction nozzle groove are set on described workbench, described chip groove is used for placing micro-fluidic immune detection chip, described reagent trough is used for placing reagent bottle, and described suction nozzle groove is used for placing suction nozzle and assembles for working metal pipe.

3. automated analysis instrument according to claim 2, is characterized in that, described chip groove, reagent trough and suction nozzle groove are arranged side by side successively, and after described suction nozzle groove, waste liquid tank is set.

4. automated analysis instrument according to claim 3, is characterized in that, the edge of described waste liquid tank arranges suction nozzle draw-in groove, supplies the card of suction nozzle on working metal pipe to fall.

5. according to the automated analysis instrument described in claim 1-4 any one, it is characterized in that, described liquid pipettes syringe pump and the conduit that module comprises stepper motor and described step motor control, and described syringe pump is communicated with described working metal pipe by conduit.

6. according to the automated analysis instrument described in claim 1-5 any one, it is characterized in that, described detection module is CCD camera, and described CCD camera is fixed on described three-dimensional machinery arm, with the motion of described three-dimensional machinery arm, moves.

7. according to the automated analysis instrument described in claim 1-6 any one, it is characterized in that, described three-dimensional machinery arm consists of the cover of three in X, Y and Z-direction servomotor, leading screw and guide rail.

8. according to the automated analysis instrument described in claim 1-7 any one, it is characterized in that, described apparatus frame is light closed structure;

Preferably, described apparatus frame mainly consists of metal framework and sheet metal.

9. according to the automated analysis instrument described in claim 1-8 any one, it is characterized in that, described CCD camera connects computing machine, and described computing machine receives and process the chemiluminescence signal of described CCD camera transmission.

10. for an automatic analysis system for micro-fluidic immune detection chip, comprise automated analysis instrument as described in claim 1-9 any one and the computing machine of connection thereof.

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