High-flux microfluidic analysis device

Abstract

The invention provides a high-flux micro-fluidic analysis device which comprises a shell, a micro-fluidic disc, a fluorescence detection module and a driving module, wherein the micro-fluidic disc, the fluorescence detection module and the driving module are mounted in the shell; the microfluidic disc comprises a central positioning hole and a plurality of liquid inlet areas distributed around the central positioning hole, each liquid inlet area comprises a sample inlet hole, the sample inlet hole is communicated with an elution quantitative hole through a first liquid channel, the elution quantitative hole is communicated with a waste liquid hole through a second liquid channel, the second liquid channel is communicated with a third liquid channel through a capillary tube, and the third liquid channel is communicated with a second liquid channel through a capillary tube. The third liquid channel is connected with a plurality of detection holes; the driving module comprises a stepping motor, and a micro-fluidic disc fixing seat is mounted on an upper output shaft of the stepping motor. According to the high-throughput microfluidic analysis device, sample treatment, reaction, analysis and the like are integrated, the sample elution and quantitative sample injection process can be rapidly completed through the microfluidic disc, the sample treatment time is greatly shortened, and the detection efficiency is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of biological sample detection, and in particular relates to a high-throughput microfluidic analysis device. Background Art

[0002] A high-throughput microfluidic analysis device is an experimental device based on microfluidic technology, which has the advantages of high efficiency, precision, miniaturization and reagent saving. A high-throughput microfluidic analysis device is usually composed of a housing, a microfluidic disk, a fluorescence detection module and a drive module, including sample reaction, separation, detection and other functions. It can process a large number of samples at the same time, significantly improve experimental efficiency and reduce experimental costs. Therefore, it is widely used in drug screening, genomics, proteomics, environmental monitoring and food safety. In the prior art, the microfluidic disk of the high-throughput microfluidic analysis device is not convenient for eluting and enriching magnetically absorbed microbial samples, and cannot be multi-channel quantitatively injected, which affects the detection efficiency. Summary of the invention

[0003] In view of the technical problems in the prior art that the microfluidic disk of the high-throughput microfluidic analysis device is not convenient for eluting magnetically absorbed microbial samples and cannot perform multi-channel quantitative sampling, the present invention proposes a high-throughput microfluidic analysis device that integrates sample processing, reaction and analysis to improve detection efficiency.

[0004] In order to achieve the above object, the technical solution adopted by the present invention is:

[0005] A high-throughput microfluidic analysis device comprises a housing and a microfluidic disk, a fluorescence detection module and a driving module installed in the housing;

[0006] The microfluidic disk includes a central positioning hole and a plurality of liquid inlet areas distributed around the central positioning hole, wherein the liquid inlet area includes a sample injection hole, the sample injection hole is connected to the elution quantitative hole via a first liquid channel, the elution quantitative hole is connected to the waste liquid hole via a second liquid channel, the second liquid channel is connected to the third liquid channel via a capillary, and the third liquid channel is connected to a plurality of detection holes;

[0007] The driving module comprises a stepping motor, and a microfluidic disk fixing seat is installed on the upper output shaft of the stepping motor.

[0008] Preferably, the housing comprises a box body and an upper cover hinged to the box body, and a flip mechanism is connected between the box body and the upper cover.

[0009] Preferably, the flip cover mechanism includes a reduction motor fixed in the box body, the output shaft of the reduction motor is connected to the first rocker, a second rocker is arranged on the inner side of the upper cover, a connecting rod is hinged between the first rocker and the second rocker, and the reduction motor rotates forward and reverse to realize the opening and closing of the upper cover.

[0010] Preferably, a cover opening limit switch and a cover closing limit switch are installed in the box body.

[0011] Preferably, the driving module comprises a mounting frame, on which a capacitive proximity switch for detecting the microfluidic disk is arranged.

[0012] Preferably, the driving module includes a shading plate and a magnetic encoder, the shading plate and the rotor of the magnetic encoder are mounted on the lower output shaft of the stepper motor, a slot-type optical coupler cooperating with the shading plate is mounted on the mounting frame, and a radial magnet of the magnetic encoder is installed under the stepper motor.

[0013] Preferably, the liquid inlet area is arranged into four groups, each group comprising 7 detection holes.

[0014] Preferably, the liquid inlet area further comprises a sample redundant hole, and the sample redundant hole is connected to the elution quantitative hole via a fourth liquid channel.

[0015] Preferably, a temperature control module is further included, wherein the temperature control module includes an upper heating plate installed on the inner side of the upper cover and a lower heating plate installed on the microfluidic plate fixing seat.

[0016] Preferably, a controller is also included, and the stepper motor, reduction motor, upper heating plate, lower heating plate, capacitive proximity switch, magnetic encoder, slot-type optical coupler, cover opening limit switch and cover closing limit switch are all electrically connected to the controller.

[0017] Compared with the prior art, the advantages and positive effects of the present invention are:

[0018] 1. The high-throughput microfluidic analysis device of the present invention adopts an innovatively designed microfluidic disk, which only requires a very small amount of sample and reagent to complete the test, and can reduce the experimental cost. The microfluidic disk can quickly complete the sample elution and quantitative injection process, greatly shortening the sample processing time, especially the sample preparation and reaction time, and improving the detection efficiency.

[0019] 2. The high-throughput microfluidic analysis device of the present invention has a high degree of integration and automation, integrating sample processing, reaction and analysis, reducing the risks of sample transmission and cross contamination, and improving the automation level of the experiment.

[0020] 3. The high-throughput microfluidic analysis device of the present invention is provided with a temperature control module, which can realize dual amplification of target sequence and signal under isothermal conditions by isothermal amplification, thereby greatly improving the sensitivity of detection and being applicable to the detection of low-concentration targets.

[0021] 4. The equipment is integrated as a whole, small in size, easy to carry, and suitable for on-site detection and instant analysis. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Schematic diagram of the overall structure of the high-throughput microfluidic analysis device of the present invention;

[0023] Figure 2 Schematic diagram of the microfluidic disk structure of the high-throughput microfluidic analysis device of the present invention;

[0024] Figure 3 A cross-sectional view of a microfluidic disk of a high-throughput microfluidic analysis device of the present invention;

[0025] Figure 4 Schematic diagram of the structure of the driving module of the high-throughput microfluidic analysis device of the present invention;

[0026] Figure 5 for Figure 4 Section view in the AA direction;

[0027] Figure 6 It is a schematic diagram of the structure of the flip cover mechanism of the high-throughput microfluidic analysis device of the present invention;

[0028] In the above figures: 1, shell; 11, upper cover; 12, box; 2, microfluidic disk; 21, central positioning hole; 211, positioning groove; 22, injection hole; 221, first liquid channel; 23, elution quantitative hole; 231, second liquid channel; 24, waste liquid hole; 25, detection hole; 251, third liquid channel; 252, capillary; 26, sample redundant hole; 261, fourth liquid channel; 27, first exhaust hole; 28, second exhaust hole; 3, fluorescence detection module; 4, drive module; 4 1. Mounting frame; 42. Stepper motor; 43. Sunshade; 44. Magnetic encoder; 45. Microfluidic disk fixing seat; 46. Capacitive proximity switch; 47. Slot-type optical coupler; 5. Temperature control module; 51. Upper heating plate; 52. Lower heating plate; 521. Proximity switch hole; 522. Fluorescence detection hole; 6. Circuit board; 7. Flip mechanism; 71. Gear motor; 72. First rocker; 73. Second rocker; 74. Connecting rod; 75. Open cover limit switch; 76. Close cover limit switch; 8. Polyimide film. DETAILED DESCRIPTION

[0029] In order to better understand the present invention, a detailed description is given below in conjunction with the accompanying drawings and embodiments.

[0030] Example: Figure 1 As shown, a high-throughput microfluidic analysis device includes a housing 1 and a circuit board 6 installed in the housing 1, a microfluidic disk 2, a fluorescence detection module 3, a drive module 4 and a temperature control module 5, wherein a controller is integrated on the circuit board 6;

[0031] like Figure 2 , Figure 3As shown, the microfluidic disk 2 is made of acrylic material, and its structure includes a central positioning hole 21 and multiple liquid inlet areas distributed around the central positioning hole 21. In this embodiment, four liquid inlet areas are set. A positioning groove 211 is set on one side of the central positioning hole 21, and the positioning groove 211 can be used to distinguish the installation direction for fast and correct installation.

[0032] The liquid inlet area includes an injection hole 22, an elution quantitative hole 23 and a waste liquid hole 24 that spread outward from the center of the circle, the injection hole 22 is connected to the elution quantitative hole 23 through the first liquid channel 221, the elution quantitative hole 23 is connected to the waste liquid hole 24 through the second liquid channel 231, the second liquid channel 231 is connected to the third liquid channel 251 through the capillary 252, and the third liquid channel 251 is sequentially connected to the seven detection holes 25 and the first exhaust hole 27 through the capillary 252. The liquid inlet area also includes a sample redundant hole 26, and the elution quantitative hole 23 is connected to the fourth liquid channel 261 and the second exhaust hole 28 through the sample redundant hole 26. When in use, first paste the polyimide film 8 (PDMS) on the lower surface of the microfluidic disk 2, and then pre-embed the primers to be detected in the liquid inlet. After the primers are pre-embedded, paste the polyimide film 8 (PDMS) on the upper surface. After that, holes are opened at all exhaust holes and an opening with a length of 1 cm is cut at the liquid inlet to facilitate the addition of sample liquid and other reagents.

[0033] like Figure 4 , Figure 5 As shown, the driving module 4 includes a mounting frame 41, a stepper motor 42, a shading plate 43 and a magnetic encoder 44. The stepper motor 42 is fixed on the mounting frame 41. A microfluidic disk fixing seat 45 is installed on the upper output shaft of the stepper motor 42. The upper end shape of the microfluidic disk fixing seat 45 matches the central positioning hole 21 of the microfluidic disk 2. A capacitive proximity switch 46 for detecting the microfluidic disk 2 is also provided on the mounting frame 41. The capacitive proximity switch 46 is electrically connected to the controller and is used to detect whether the microfluidic disk 2 is installed on the microfluidic disk fixing seat 45, and then the controller controls the switch of the upper cover 11 according to the detection signal.

[0034] The shading plate 43 and the rotor of the magnetic encoder 44 are mounted on the lower output shaft of the stepper motor 42, and rotate with the stepper motor 42. At the same time, a slotted optical coupler 47 that cooperates with the shading plate 43 is mounted on the mounting frame 41, and a radial magnet of the magnetic encoder 44 is mounted below the stepper motor 42. The stepper motor 42, the magnetic encoder 44 and the slotted optical coupler 47 are all electrically connected to the controller, and the stepper motor 42 controls the rotation angle and speed through the controller. After the high-throughput microfluidic analysis device starts to be powered on, the zero position is first calibrated, that is, when the shading plate 43 rotates with the stepper motor 42 and enters the gap of the slotted optical coupler 47, the slotted optical coupler 47 transmits the detection signal to the controller, and the controller controls the stepper motor 42 to stop rotating. At this time, the microfluidic disk 2 is at the zero position, and the sample liquid can be added at this time.

[0035] The temperature control module 5 includes an upper heating plate 51 mounted on the inner side of the upper cover 11 and a lower heating plate 52 mounted on the microfluidic plate fixing seat 45. The lower heating plate 52 is fixed on the mounting frame 41 and does not rotate with the microfluidic plate fixing seat 45. The lower heating plate 52 is provided with a proximity switch hole 521 and a fluorescence detection hole 522. When the microfluidic plate 2 is at the zero position, the capacitive proximity switch 46 is located directly below the proximity switch hole 521, and the microfluidic plate 2 can be detected. The upper heating plate 51 and the lower heating plate 52 are both electrically connected to the controller. The temperature control module 5 can make the microfluidic plate in a relatively constant temperature environment, and the nucleic acid rapid isothermal detection amplification technology (RIDA) can be implemented on the sample to achieve dual amplification of the target sequence and the signal under isothermal conditions, thereby greatly improving the sensitivity and specificity of the detection.

[0036] The fluorescence detection module 3 is fixed on the mounting frame 41 and is located below the lower heating plate 52. When the microfluidic plate 2 is at the zero position, the detection point is aligned with the fluorescence detection hole 522 of the lower heating plate 52. The fluorescence detection module 3 uses an integrated transceiver fluorescence detector, which sends light with a wavelength of 470um. When the light irradiates the medium of the sample and primer reaction, fluorescence is generated. The fluorescence is detected by the detection module, converted into a voltage value and transmitted to the controller to obtain a detection value. It has the advantages of high sensitivity, good selectivity, and fast response speed.

[0037] like Figure 6As shown, the housing 1 includes a box body 12 and an upper cover 11 hinged on the box body 12, and a flip mechanism 7 is connected between the box body 12 and the upper cover 11 to realize automatic opening and closing of the upper cover 11. The flip mechanism 7 includes a reduction motor 71 fixed in the box body 12, the output shaft of the reduction motor 71 is connected to the first rocker 72, a second rocker 73 is arranged inside the upper cover 11, and a connecting rod 74 is hinged between the first rocker 72 and the second rocker 73. A cover opening limit switch 75 and a cover closing limit switch 76 are also installed in the box body 12, and the cover opening limit switch 75, the cover closing limit switch 76 and the reduction motor 71 are all electrically connected to the controller, and the controller controls the reduction motor 71 to rotate forward and reverse to realize the opening and closing of the upper cover 11. When the upper cover 11 is opened to the maximum angle, the cover opening limit switch 75 is triggered, and the reduction motor 71 stops rotating. When the upper cover 11 is completely closed, the cover closing limit switch 76 is triggered to complete the cover closing.

[0038] The detection process of the high-throughput microfluidic analysis device is as follows:

[0039] The controller controls the reduction motor 71 to rotate so as to open the upper cover 11, and the prepared microfluidic disk 2 is placed on the microfluidic disk fixing seat 45. The preparation work includes pre-embedded primers in the detection holes 25, pasting transparent substrate materials on both sides, and opening exhaust holes and liquid filling ports on the transparent substrate materials. A small amount of sample is added to the four liquid inlets respectively, and the upper cover 11 is closed. The stepper motor 42 starts to rotate for centrifugal treatment at 1000r / min. After the centrifugal treatment is completed, the upper cover 11 is opened again, and a small amount of sterile water is added to the four liquid inlets respectively, and the upper cover 11 is closed; the stepper motor 42 rotates at a low and high speed. After the rotation is completed, the fluorescence detection module 3 measures the 28 detection holes 25 in turn. During the detection process, the stepper motor 42 cooperates with the magnetic encoder to rotate a fixed angle so that the detection holes 25 are aligned with the fluorescence detection module 3 in turn, thus completing one detection. The above steps are repeated according to the number of detections set by the user to complete the entire detection process.

[0040] During the detection process, a maximum of 300 ml of sample liquid can be added to the liquid inlet area of ​​the microfluidic disk 2 at a time. The process of adding samples is to first use a program-controlled motor to drive the microfluidic disk 2 to rotate so that the microfluidic disk 2 is at the zero position. In this embodiment, a liquid adding hole is provided on the upper cover 11, so liquid can be added directly from the liquid adding hole without opening the upper cover 11. Of course, the upper cover 11 can also be opened to directly inject the sample liquid. Thereafter, the sample in the liquid inlet hole is centrifuged into the elution quantitative hole 23, and after adding pure water, it is centrifuged again. The fine magnetic beads with adsorbed bacteria are centrifuged into the waste liquid hole 24, and the bacteria are eluted and squeezed into the elution quantitative hole 23. Under the action of the suction force generated by the capillary 252 and the centrifugation, the bacteria that have been eluted in the elution quantitative hole 23 are quantitatively transported to the detection hole 25 to react with the primers pre-embedded in the detection hole 25. When the detection holes 25 are filled with sample liquid, the excess liquid enters the sample remaining hole. By checking whether there is sample liquid in the sample remaining hole, it can be quickly determined whether the current added sample liquid volume is sufficient.

[0041] The high-throughput microfluidic analysis device of the present invention adopts an innovatively designed microfluidic disk 2, which can complete the detection with only a very small amount of sample and reagent, and can reduce the experimental cost. The microfluidic disk 2 can quickly complete the sample elution and quantitative injection process, greatly shortening the sample processing time, especially the sample preparation and reaction time, and improving the detection efficiency.

[0042] The high-throughput microfluidic analysis device of the present invention has a high degree of integration and automation, and integrates sample processing, reaction and analysis, etc., reducing the risk of sample transmission and cross contamination, and improving the automation level of the experiment. It is provided with a temperature control module 5, and isothermal amplification can be used to achieve dual amplification of the target sequence and the signal under isothermal conditions, thereby greatly improving the sensitivity of detection, and can be applied to the detection of low-concentration targets. The device is integrated as a whole, small in size, easy to carry, and suitable for on-site detection and instant analysis.

[0043] The above description is only a preferred embodiment of the present invention and does not limit the present invention in other forms. Any technician familiar with the profession may use the technical content disclosed above to change or modify it into an equivalent embodiment with equivalent changes and apply it to other fields. However, any simple modification, equivalent change and modification made to the above embodiment based on the technical essence of the present invention without departing from the content of the technical solution of the present invention still falls within the protection scope of the technical solution of the present invention.

Claims

1. A high-throughput microfluidic analysis device, characterized in that: It includes a housing and a microfluidic disk, a fluorescence detection module and a driving module installed in the housing; The microfluidic disk includes a central positioning hole and a plurality of liquid inlet areas distributed around the central positioning hole, wherein the liquid inlet area includes a sample injection hole, the sample injection hole is connected to the elution quantitative hole via a first liquid channel, the elution quantitative hole is connected to the waste liquid hole via a second liquid channel, the second liquid channel is connected to the third liquid channel via a capillary, and the third liquid channel is connected to a plurality of detection holes; The driving module comprises a stepping motor, and a microfluidic disk fixing seat is installed on the upper output shaft of the stepping motor.

2. The high-throughput microfluidic analysis device according to claim 1, characterized in that: The shell comprises a box body and an upper cover hinged on the box body, and a flip mechanism is connected between the box body and the upper cover.

3. The high-throughput microfluidic analysis device according to claim 2, characterized in that: The flip cover mechanism includes a reduction motor fixed in the box body, the output shaft of the reduction motor is connected to the first rocker, a second rocker is arranged on the inner side of the upper cover, a connecting rod is hinged between the first rocker and the second rocker, and the reduction motor rotates forward and reverse to realize the upper cover opening and closing.

4. The high-throughput microfluidic analysis device according to claim 3, characterized in that: A cover opening limit switch and a cover closing limit switch are installed in the box body.

5. The high-throughput microfluidic analysis device according to claim 4, characterized in that: The driving module comprises a mounting frame, on which a capacitive proximity switch for detecting the microfluidic disk is arranged.

6. The high-throughput microfluidic analysis device according to claim 5, characterized in that: The driving module includes a shading plate and a magnetic encoder. The shading plate and the rotor of the magnetic encoder are installed on the lower output shaft of the stepper motor. A slot-type optical coupler matching the shading plate is installed on the mounting frame. A radial magnet of the magnetic encoder is installed below the stepper motor.

7. The high-throughput microfluidic analysis device according to claim 1, characterized in that: The liquid inlet area is arranged in four groups, each group including 7 detection holes.

8. The high-throughput microfluidic analysis device according to claim 1, characterized in that: The liquid inlet area also includes a sample redundant hole, and the sample redundant hole is connected to the elution quantitative hole through a fourth liquid channel.

9. The high-throughput microfluidic analysis device according to claim 6, characterized in that: It also includes a temperature control module, which includes an upper heating plate installed on the inner side of the upper cover and a lower heating plate sleeved on the microfluidic plate fixing seat.

10. The high-throughput microfluidic analysis device according to claim 9, characterized in that: It also includes a controller, and the stepper motor, reduction motor, upper heating plate, lower heating plate, capacitive proximity switch, magnetic encoder, slot-type optical coupler, cover opening limit switch and cover closing limit switch are all electrically connected to the controller.

Images