CN109946460B - Chemiluminescence immune detection device based on micro-fluidic chip and using method - Google Patents
Chemiluminescence immune detection device based on micro-fluidic chip and using method Download PDFInfo
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Abstract
The invention provides a chemiluminescence immunoassay device based on a microfluidic chip and a use method thereof, belonging to the field of biochemical inspection equipment, and comprising a reaction chamber, the microfluidic chip is arranged in the reaction chamber and driven to rotate by a rotating motor, a lifting assembly is used for driving the rotating motor to lift up and down so as to realize the switching of chip washing, reaction and acquisition positions, a magnetic ring is arranged at the lower end inside the reaction chamber and used for absorbing magnetic beads to provide continuous magnetic attraction when the chip rotates at a high speed, an acquisition module is used for acquiring optical signals and uploading the optical signals to a control panel, a temperature control assembly is used for controlling the temperature of the reaction chamber so as to meet the enzyme reaction requirement, and the control panel, the temperature control assembly and the action of the rotating motor are controlled to control the rotation and stop of the microfluidic chip. The invention has high integration level, simple and convenient operation and realizes full-automatic detection.
Description
Technical Field
The invention belongs to the field of biochemical inspection equipment, and relates to a chemiluminescence immunoassay device based on a microfluidic chip and a use method thereof.
Background
The micro-fluidic chip technology is a new technology for accurately manipulating and controlling nano-liter and pico-liter fluid (biological sample fluid) in a micrometer-scale runner, and by using the technology, basic operation units such as sample preparation, reaction, separation, detection, cell culture, separation, cracking and the like related in the fields of chemistry, biology and the like can be integrated or basically integrated on a chip with a few square centimeters (even smaller), and a network is formed by the micro-runner so as to control the fluid to penetrate through the whole system and replace a technical platform with various functions of a conventional chemistry or biology laboratory. The basic characteristics and the greatest advantages of the microfluidic chip laboratory are that a plurality of unit technologies are flexibly combined and integrated on a small platform with controllable whole. .
Chemiluminescent immunoassay (chemiluminescence immunoassay, CLIA) combines a chemiluminescent assay technique with high sensitivity with a highly specific immune reaction, and is used for detection and analysis of various antigens, haptens, antibodies, hormones, enzymes, fatty acids, vitamins, drugs, etc. Is a latest immunoassay technology developed after the analysis of radioimmunoassay, enzyme immunoassay, fluorescence immunoassay and time-resolved fluorescence immunoassay.
Chemiluminescent immunoassay methods are basically classified into the following three types:
1) Direct chemiluminescence, wherein the label is an acridinium ester;
2) Enzymatic chemiluminescence, wherein the label is alkaline phosphatase or horseradish peroxidase;
3) Electrochemiluminescence, wherein the marker is terpyridyl ruthenium;
at present, related equipment based on microfluidic chips and capable of detecting multiple items at the same time does not exist in the market, and after the production of the double-antibody sandwich compound, in the process of washing and removing unbound enzyme-labeled antibodies, a method adopted by the traditional large-scale detection equipment adopts a mode of multiple absorption, dilution and absorption, so that the flow is complex and more waste liquid is generated.
Disclosure of Invention
The invention aims to solve the problem of providing a chemiluminescence immunoassay device based on a microfluidic chip and a use method thereof, and the chemiluminescence immunoassay device has the advantages of high integration degree, simplicity and convenience in operation and realization of full-automatic detection.
In order to solve the technical problems, the invention adopts the following technical scheme: the chemiluminescence immunoassay device based on the microfluidic chip comprises a reaction chamber, and the microfluidic chip is arranged in the reaction chamber and driven to rotate by a rotating motor;
the lifting assembly is used for driving the rotating motor to lift up and down so as to realize the switching of chip washing, reaction and acquisition positions;
the magnetic ring is arranged at the lower end of the inside of the reaction chamber and is used for adsorbing magnetic beads to provide continuous magnetic attraction when the chip rotates at a high speed;
the acquisition module is used for acquiring the optical signals and uploading the optical signals to the control panel;
the temperature control component is used for controlling the temperature of the reaction chamber and meeting the enzyme reaction requirement;
and the control panel is used for controlling the actions of the acquisition module, the temperature control assembly and the rotating motor and controlling the rotation and stop of the microfluidic chip.
Further, the lifting assembly is of a scissor type lifting table structure, the lifting assembly comprises a lifting table with the upper end horizontally arranged, the rotating motor is fixedly arranged on the lifting table, the lifting assembly is driven by the lifting motor, and the lifting motor is controlled to rotate by the control panel.
Further, the lifting assembly further comprises a base, the structure of the base is the same as that of the lifting platform and is arranged symmetrically up and down, a first connecting rod, a second connecting rod and a rotating shaft which form a scissor assembly are arranged between the base and the lifting platform, the lower end of the scissor assembly is in sliding connection with the base through a sliding block, the upper end of the scissor assembly is provided with a lead screw and a screw driving relative lifting platform which are in sliding connection, the lead screw is driven to rotate by a lifting motor, and the screw is in sliding connection with the lifting platform through a guide rod erected at the lower end of the lifting platform.
Further, collection module includes balance frame, collector and sealing member, reaction chamber's upper end is equipped with the collection window, the balance frame is in the upper end of sealed cavity, the sealing member is established the lower extreme of balance frame, the sealing member is flexible material, under the collection state, microfluidic chip upwards move with the sealing member contact sets up, the sealing member is established collection window's outer lane, the collector is established the upper end of balance frame and correspond collection window sets up.
Further, three collection windows are arranged at the upper end of the reaction chamber, and sealing rings are arranged at the positions, corresponding to the collection windows, of the lower end of the balancing stand.
Further, the control by temperature change subassembly is including establishing at reaction chamber rear end and the U-shaped wind channel of its inside intercommunication, the one end in U-shaped wind channel is as the air intake, and the other end is as the air outlet, and inside middle part is equipped with heating element, air intake and air outlet are close to reaction chamber department all is equipped with axial fan, the control by temperature change subassembly is still including setting up the temperature collector in reaction chamber, the temperature collector with the control panel electricity is connected.
Further, the microfluidic chip comprises a diluent sample adding hole, a sample adding hole, a detection groove and a waste liquid groove, wherein the diluent sample adding hole and the sample adding hole are communicated with the mixing groove, the mixing groove is communicated with the detection groove, the lower end of the detection groove is provided with a cleaning liquid quantitative groove communicated with the detection groove, and the detection groove is communicated with the waste liquid groove.
The method for using the chemiluminescence immune detection device based on the micro-fluidic chip comprises the following steps of S1, equipment starting preparation;
s2, adding a detection sample to a sample adding hole, adding diluent to a diluent adding hole of the microfluidic chip, and placing the microfluidic chip after adding into a reaction cavity and fixing the microfluidic chip with a rotating motor;
s3: after the installation is finished, the rotating motor and the microfluidic chip are located at a reaction position after descending, the rotating motor drives the microfluidic chip to rotate at a high speed, and the microfluidic chip is matched to sequentially finish the separation, quantification, dilution and heating of a sample and a diluent; enabling the mixed solution of the quantified sample and the diluent to enter a detection groove of the microfluidic chip, dissolving the magnetic bead antibody freeze-dried pellets and the enzyme-labeled antibody freeze-dried pellets pre-installed in the detection groove, and enabling the magnetic bead antibody freeze-dried pellets and the enzyme-labeled antibody freeze-dried pellets to react to form a double-antibody sandwich compound;
s4, the lifting assembly descends to enable the microfluidic chip to reach a washing position, magnetic beads are attached to the bottom surface of a chip detection groove under the action of magnetic force of the magnetic ring, the rotating motor rotates at a high speed, liquid containing redundant enzyme-labeled antibodies enters the waste liquid groove under the action of the centrifugal force, and double-antibody sandwich compound is reserved in the detection groove under the action of the magnetic attraction;
s5, lifting the lifting assembly to enable the magnetic beads to be separated from the magnetic force constraint of the magnetic ring, releasing the cleaning liquid in the cleaning liquid quantitative tank, enabling the cleaning liquid to enter the detection tank by rotation of the motor, and enabling the magnetic beads to be resuspended;
s6, returning the rotating motor and the microfluidic chip to a reaction position, releasing the luminescent substrate, rotating the microfluidic chip to enable the luminescent substrate to enter a detection groove, and completing mixing and reaction of detection liquid by matching with the microfluidic chip;
and S7, the rotating motor drives the microfluidic chip to rotate, so that the detection groove is aligned with the acquisition module, and the acquired optical signals are uploaded to the control panel for quantitative analysis.
Further, the control temperature of the temperature control component is 37 degrees + -0.3 degrees.
Further, repeating the steps 4, 5 and 4 between S5 and S, washing with the washing liquid for 2 times to completely eliminate the unbound enzyme-labeled antibody, and completing the washing step.
Compared with the prior art, the invention has the following advantages and positive effects.
1. The invention sets up the micro-fluidic chip driven by the motor, can realize the ration and separation of the liquid under the high-speed rotation of the motor, set up the magnetic ring at the same time, absorb the magnetic bead and offer the continuous magnetic attraction when being used for the chip to rotate at a high speed, have realized the separation and cleaning of the liquid, set up the lifting assembly for the motor at the same time, have realized the micro-fluidic chip is in the free switching of washing position, reaction position, collection position, centrifuge, quantitate, dilute, mix, wash in an organic whole of the detection sample, set up collection module and temperature control assembly at the same time, guarantee the temperature environment of the reaction is invariable, confirm the accuracy of detection, collect assembly carries on the collection and processing of the data in time, all electric components are controlled by the control panel, the degree of automation is high, the middle process is unmanned to participate, has promoted the work efficiency, has promoted the intensity of labour, and promoted the precision of detection, the degree of integration is high, easy and simple and convenient to operate, realize the full-automatic detection;
2. three collection windows are arranged at the upper end of the reaction chamber, sealing rings are arranged at the positions, corresponding to the collection windows, of the lower end of the balancing frame, when the optical signals are collected, in order to avoid light leakage caused by the fact that the chip is partially extruded and unbalanced, collection windows which are uniformly distributed relative to the axis of the microfluidic chip are added on the upper surface of the reaction chamber, and symmetrical points are formed between the support points and the collection modules so as to ensure the stress balance of the microfluidic chip;
3. the arrangement of the two axial flow fans ensures that air in the reaction chamber where the microfluidic chip is located flows, the heating component is positioned at the middle part of the U-shaped air channel, the air passing through the air channel is circularly heated under the drive of the axial flow fans, the temperature collector is arranged at the front end in the reaction chamber, the heating output proportion of the circulating air in the air channel is controlled according to the feedback of the temperature collector, the temperature of the reaction chamber is ensured to be 37+/-0.3 ℃, and the constant temperature effect is good.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention. In the drawings:
FIG. 1 is a schematic diagram of an enzymatic chemiluminescent process reaction involved in the present invention;
FIG. 2 is a schematic structural diagram of a chemiluminescent immunoassay device based on a microfluidic chip of the present invention;
FIG. 3 is a top view of a microfluidic chip-based chemiluminescent immunoassay device of the present invention;
FIG. 4 is a cross-sectional view B-B of FIG. 3 in accordance with the present invention;
FIG. 5 is a detail view of section A of FIG. 4 in accordance with the present invention;
FIG. 6 is a front view of a microfluidic chip-based chemiluminescent immunoassay device of the present invention;
FIG. 7 is a cross-sectional view of the C-C of FIG. 6 in accordance with the present invention;
FIG. 8 is a schematic structural view of a balance frame according to the present invention;
FIG. 9 is a schematic diagram of a chemiluminescent immunoassay device based on a microfluidic chip in a washing position;
FIG. 10 is a schematic structural view of a chemiluminescent immunoassay device based on a microfluidic chip in a reaction position;
FIG. 11 is a schematic diagram of a chemiluminescent immunoassay device based on a microfluidic chip in a collection position;
fig. 12 is a schematic structural view of a microfluidic chip according to the present invention.
Reference numerals:
1. a reaction chamber; 11. a collection window; 2. a lifting assembly; 21. a lifting table; 22. a base; 23. a first link; 24. a second link; 25. a rotating shaft; 26. a slide block; 27. a screw rod; 28. a nut; 3. an acquisition module; 31. a collector; 32. a balancing stand; 33. a seal; 4. a temperature control assembly; 41. u-shaped air pipes; 42. an axial flow fan; 43. a heating assembly; 45. a temperature collector; 5. a control panel; 6. a rotating electric machine; 7. a microfluidic chip; 71. a diluent sample adding hole; 72. a sample loading hole; 73. a detection groove; 74. a waste liquid tank; 75. a mixing tank; 76. a luminescent substrate storage tank; 77. a cleaning liquid quantitative tank; 8. and a magnetic ring.
Detailed Description
It should be noted that, without conflict, the embodiments of the present invention and features of the embodiments may be combined with each other.
In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first", "a second", etc. may explicitly or implicitly include one or more such feature. In the description of the present invention, unless otherwise indicated, the meaning of "a plurality" is two or more.
In the description of the present invention, it should be noted that, unless explicitly specified and limited otherwise, the terms "mounted," "connected," and "connected" are to be construed broadly, and may be either fixedly connected, detachably connected, or integrally connected, for example; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art in a specific case.
Specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings.
As shown in FIG. 1, the reaction principle of the enzymatic chemiluminescence method is as follows: and (3) putting the antibody coated with the magnetic beads, the mould-labeled antibody and the detection sample containing the antigen together for reacting for a period of time to form a double-antibody sandwich compound, washing and removing the unbound enzyme-labeled antibody, adding a quantitative luminescent substrate for reacting with the mould to emit light with a specific wavelength, and collecting the photon number by adopting a photomultiplier tube for quantitative analysis.
As shown in fig. 2 to 7, the present invention is a chemiluminescent immunoassay device based on a microfluidic chip, comprising a reaction chamber 1, and a microfluidic chip 7 disposed in the reaction chamber 1 and driven to rotate by a rotating motor 6;
the lifting assembly 2 is used for driving the rotating motor 6 to lift up and down so as to realize the washing, reaction and acquisition position switching of the microfluidic chip 7;
the magnetic ring 8 is arranged at the lower end of the inside of the reaction chamber 1 and is used for adsorbing magnetic beads to provide continuous magnetic attraction when the microfluidic core chip rotates at a high speed;
the acquisition module 3 is used for acquiring optical signals and uploading the optical signals to the control panel 5;
the temperature control component 4 is used for controlling the temperature of the reaction chamber 1 and meeting the enzyme reaction requirement;
and the control panel 5 controls the actions of the acquisition module 3, the temperature control assembly 4 and the rotating motor 6 and controls the rotation and stop of the micro-fluidic chip 7.
Preferably, the lifting assembly 2 is a scissor type lifting platform 21 structure, the lifting assembly 2 comprises a lifting platform 21 with a horizontally arranged upper end, a rotating motor 6 is fixedly arranged on the lifting platform 21, the lifting assembly 2 is driven by the lifting motor, the lifting motor is controlled by a control panel 5 to rotate, preferably, the lifting assembly 2 further comprises a base 22, the structure of the lifting platform 21 of the base 22 is the same and the two are symmetrically arranged up and down, a first connecting rod 23, a second connecting rod 24 and a rotating shaft 25 which form the scissor assembly are arranged between the base 22 and the lifting platform 21, the lower end of the scissor assembly is in sliding connection with the base 22 through a sliding block 26, the upper end of the scissor assembly is provided with a lead screw 27, the lead screw 28 is in sliding connection with the lifting platform 21, the lead screw 27 is driven to rotate by the lifting motor, the lead screw 27 is in matching arrangement with the lead screw 28, the horizontal movement of the lead screw 28 on the lead screw is realized, the cross angle change of the first connecting rod 23 and the second connecting rod 24 is realized, the lifting action is finally realized, the lifting motor is simple in structure, the lifting panel is controlled reliably, the control panel is controlled to be not to be high, the PLC is controlled to be high and the PLC is convenient to position is convenient to control the position and is convenient to use, and the program is stable to be controlled to be 5 is easy to be controlled to be high.
Preferably, the collection module 3 includes balancing stand 32, collector 31 and sealing member 33, the upper end of reaction chamber 1 is equipped with collection window 11, balancing stand 32 is established in the upper end of sealed cavity, sealing member 33 is established in the lower extreme of balancing stand 32, sealing member 33 is flexible material, under the collection state, microfluidic chip 7 upwards moves and contacts the setting with sealing member 33, sealing member 33 establishes the outer lane at collection window 11, collector 31 establishes the upper end at balancing stand 32 and corresponds collection window 11 setting, sealing member 33's setting has guaranteed the leakproofness under the collection state, guarantee signal acquisition's efficiency and accuracy.
Preferably, three collection windows 11 are arranged at the upper end of the reaction chamber 1, sealing rings are arranged at the positions, corresponding to the collection windows 11, of the lower end of the balancing frame 32, when the optical signals are collected, in order to avoid light leakage caused by the fact that the chip is out of balance due to local extrusion, and to influence the collection result, the collection windows 11 which are uniformly distributed relative to the axis of the microfluidic chip 7 are added on the upper surface of the reaction chamber 1, and symmetrical points are formed between the support points and the collection modules 3 so as to ensure that the microfluidic chip 7 is stressed and balanced.
Preferably, the temperature control assembly 4 includes a U-shaped air channel disposed at the rear end of the reaction chamber 1 and communicated with the interior thereof, one end of the U-shaped air channel is used as an air inlet, the other end is used as an air outlet, a heating assembly 43 is disposed in the middle of the interior, the air inlet and the air outlet are both provided with an axial flow fan 42 near the reaction chamber 1, the temperature control assembly 4 further includes a temperature collector 4531 disposed in the reaction chamber 1, the temperature collector 4531 is electrically connected with the control panel 5, the two axial flow fans 42 are disposed, the air in the reaction chamber 1 where the microfluidic chip 7 is disposed flows, the heating assembly 43 is disposed in the middle of the U-shaped air channel, the air passing through the air channel is circularly heated under the driving of the axial flow fan 42, the temperature collector 4531 is disposed at the front end in the reaction chamber 1, the heating output ratio of the circulating air in the air channel is controlled according to the feedback of the temperature collector 4531, the temperature of the reaction chamber 1 is guaranteed to be 37 degrees + -0.3 degrees, the constant temperature effect is good, and the heating assembly 43 can be an electric heating plate or other existing structure.
Preferably, the microfluidic chip 7 comprises a diluent sample adding hole 71, a sample adding hole 72, a detection groove 73 and a waste liquid groove 74, wherein the diluent sample adding hole 71 and the sample adding hole 72 are communicated with the mixing groove 75, the mixing groove 75 is communicated with the detection groove 73, a cleaning liquid quantitative groove 77 communicated with the detection groove 73 is arranged at the lower end of the detection groove 73, the detection groove 73 is communicated with the waste liquid groove 74, a luminous substrate storage groove 76 communicated with the detection groove 73 is further arranged at the lower end of the detection groove 73, the sample and the diluent are firstly mixed in the mixing groove 75, the sample and the diluent are rotated at a high speed and a non-uniform speed under the driving of a motor, the full fusion of the sample and the diluent is realized, then the mixed liquid enters the detection groove 73, the secondary fusion reaction is realized under the rotation of the microfluidic chip 7, the double-antibody sandwich compound is formed, the whole structure is simple, the principle of centrifugal force is fully utilized, the quantitative and fusion of the liquid are realized by combining different rotation speeds and rotation directions, the effect of the magnetic ring 8 is realized, the cleaning and the separation are realized, and the structural basis is provided for automatic chemiluminescence immunodetection.
In the actual working process, the method is carried out according to the following steps of S1, equipment is started up and prepared; s2, adding a detection sample to a sample adding hole 72, adding diluent to a diluent adding hole 71 of the microfluidic chip 7, and placing the microfluidic chip 7 with the added sample in the reaction chamber 1 and fixing the microfluidic chip with a motor, wherein the steps are performed when the lifting table 21 is at the highest point as shown in FIG. 11; s3: after the installation is completed, the rotating motor 6 and the microfluidic chip 7 are located at a reaction position after descending, as shown in fig. 10, the rotating motor 6 drives the microfluidic chip 7 to rotate at a high speed, and the separation, quantification, dilution and heating of a sample and a diluent are sequentially completed by matching the chips; the mixed solution of the quantified sample and the diluent enters a detection groove 73 of the microfluidic chip 7, and the magnetic bead antibody freeze-dried pellets and the enzyme-labeled antibody freeze-dried pellets preloaded in the detection groove are dissolved and reacted to form a double-antibody sandwich compound; s4, the lifting assembly 2 descends to enable the microfluidic chip 7 to reach a washing position, as shown in FIG. 9, magnetic beads are attached to the bottom surface of the chip detection groove 73 under the action of magnetic force of the magnetic ring 8, the rotating motor rotates at a high speed, liquid containing redundant enzyme-labeled antibodies enters the waste liquid groove 74 under the action of centrifugal force, and the double-antibody sandwich compound is reserved in the detection groove 73 under the action of magnetic attraction; s5, lifting the lifting assembly 2 to enable the magnetic beads to be separated from the magnetic force constraint of the magnetic ring 8, releasing the cleaning liquid in the cleaning liquid quantitative tank, enabling the cleaning liquid to enter the detection tank 73 by rotating the motor, and re-suspending the magnetic beads; s6, repeating the steps 4, 5 and 4, washing with the cleaning liquid for 2 times, thoroughly removing the unbound enzyme-labeled antibody, and finishing the washing step; s7, returning the rotating motor and the microfluidic chip to a reaction position, releasing the luminescent substrate, rotating the microfluidic chip 7 to enable the luminescent substrate to enter the detection groove 73, and completing mixing and reaction of detection liquid by matching with the microfluidic chip 7; and S8, the motor drives the micro-fluidic chip 7 to rotate, so that the detection groove 73 is aligned with the acquisition window 11, the lifting assembly 2 is lifted to the acquisition position, is contacted with the sealing piece 33 and is compressed and deformed, and the tightness is ensured, so that a closed cavity is formed with the acquisition window 11 for optical signal acquisition, the acquired optical signal is uploaded to the control panel 5 by the collector 31 for quantitative analysis, the detection principle is that the structure improves the detection convenience.
In the whole process, the temperature is controlled to be 37+/-0.3 ℃ through the temperature control assembly 4, the structure of the temperature control assembly 4 is simple, the heating mode has the advantages of saving energy consumption, high temperature control precision and temperature uniformity in the whole chamber, the whole structure is matched with the micro-fluidic chip 7 through the magnetic ring 8, under the condition of high-speed rotation, the washing step of enzymatic chemiluminescence is realized through a centrifugal mode, the processing of a sample and the detection of the enzymatic chemiluminescence method are completed through the matched chip, the free switching of the washing position, the reaction position and the acquisition position of the micro-fluidic chip 7 is realized through the matched lifting assembly 2, the micro-fluidic chip 7 is driven to be centrifuged at a high speed, the liquid is mixed in a positive and negative rotation mode, the accurate position is stopped, the stable operation of the chip is ensured, the structure is compact, the operation is simple, the operation is reliable, the rapid detection is carried out, the degree of automation is high, the working efficiency is improved, and the detection precision is ensured.
The foregoing describes one embodiment of the present invention in detail, but the description is only a preferred embodiment of the present invention and should not be construed as limiting the scope of the invention. All equivalent changes and modifications within the scope of the present invention are intended to be covered by the present invention.
Claims (4)
1. Chemiluminescence immunoassay device based on micro-fluidic chip, its characterized in that: the micro-fluidic chip is arranged in the reaction chamber and driven to rotate by a rotating motor;
the lifting assembly is used for driving the rotating motor to lift up and down so as to realize the switching of the washing, reaction and acquisition positions of the microfluidic chip;
the magnetic ring is arranged at the lower end of the inside of the reaction chamber and is used for adsorbing magnetic beads to provide continuous magnetic attraction when the microfluidic chip rotates at a high speed;
the acquisition module is used for acquiring the optical signals and uploading the optical signals to the control panel;
the temperature control component is used for controlling the temperature of the reaction chamber and meeting the enzyme reaction requirement;
the control panel is used for controlling the actions of the acquisition module, the temperature control assembly and the rotating motor and controlling the rotation and stop of the microfluidic chip;
the collection module comprises a balance frame, a collector and a sealing element, wherein the upper end of the reaction chamber is provided with a collection window, the balance frame is arranged at the upper end of the reaction chamber, the sealing element is arranged at the lower end of the balance frame and is made of flexible materials, the microfluidic chip moves upwards to be in contact with the sealing element in a collection state, the sealing element is arranged at the outer ring of the collection window, and the collector is arranged at the upper end of the balance frame and is arranged corresponding to the collection window;
the microfluidic chip comprises a diluent sample adding hole, a sample adding hole, a detection groove and a waste liquid groove, wherein the diluent sample adding hole and the sample adding hole are communicated with a mixing groove, the mixing groove is communicated with the detection groove, the lower end of the detection groove is provided with a cleaning liquid quantitative groove communicated with the detection groove, and the detection groove is communicated with the waste liquid groove;
the temperature control assembly comprises a U-shaped air duct which is arranged at the rear end of the reaction chamber and is communicated with the rear end of the reaction chamber, one end of the U-shaped air duct is used as an air inlet, the other end of the U-shaped air duct is used as an air outlet, a heating assembly is arranged in the middle of the interior of the U-shaped air duct, the air inlet and the air outlet are close to the reaction chamber, an axial flow fan is arranged at the reaction chamber, the temperature control assembly further comprises a temperature collector which is arranged in the reaction chamber, and the temperature collector is electrically connected with the control panel.
2. The microfluidic chip-based chemiluminescent immunoassay device of claim 1 wherein: the upper end of the reaction chamber is provided with three collection windows, and the lower end of the balancing stand and the position corresponding to the collection windows are provided with sealing rings.
3. A method of using the microfluidic chip-based chemiluminescent immunoassay device of claim 1 characterized by:
s1, equipment is started up and prepared;
s2, adding a detection sample to a sample adding hole, adding diluent to a diluent adding hole of the microfluidic chip, and placing the microfluidic chip after adding into a reaction cavity and fixing the microfluidic chip with a rotating motor;
s3: the rotating motor and the microfluidic chip are positioned at a reaction position after descending, the rotating motor drives the microfluidic chip to rotate at a high speed, and the microfluidic chip is matched to sequentially finish separation, quantification, dilution and heating of a sample and a diluent; enabling the mixed solution of the quantified sample and the diluent to enter a detection groove of the microfluidic chip, dissolving the magnetic bead antibody freeze-dried pellets and the enzyme-labeled antibody freeze-dried pellets pre-installed in the detection groove, and enabling the magnetic bead antibody freeze-dried pellets and the enzyme-labeled antibody freeze-dried pellets to react to form a double-antibody sandwich compound;
s4, the lifting assembly descends to enable the microfluidic chip to reach a washing position, magnetic beads are attached to the bottom surface of a chip detection groove under the action of magnetic force of the magnetic ring, the rotating motor rotates at a high speed, liquid containing redundant enzyme-labeled antibodies enters the waste liquid groove under the action of the centrifugal force, and double-antibody sandwich compound is reserved in the detection groove under the action of the magnetic attraction;
s5, lifting the lifting assembly to enable the magnetic beads to be separated from the magnetic force constraint of the magnetic ring, releasing the cleaning liquid in the cleaning liquid quantitative tank, enabling the cleaning liquid to enter the detection tank by rotation of the motor, and enabling the magnetic beads to be resuspended;
s6, repeating the steps 4, 5 and 4, washing with the cleaning liquid for 2 times, thoroughly removing the unbound enzyme-labeled antibody, and finishing the washing step;
s7, returning the rotating motor and the microfluidic chip to a reaction position, releasing the luminescent substrate, rotating the microfluidic chip to enable the luminescent substrate to enter a detection groove, and completing mixing and reaction of detection liquid by matching with the microfluidic chip;
and S8, the rotating motor drives the microfluidic chip to rotate, so that the detection groove is aligned with the acquisition module, and the acquired optical signals are uploaded to the control panel for quantitative analysis.
4. The method for using a microfluidic chip-based chemiluminescent immunoassay device of claim 3 wherein: the control temperature of the temperature control component is 37 degrees + -0.3 degrees.
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| CN201910313140.7A CN109946460B (en) | 2019-04-18 | 2019-04-18 | Chemiluminescence immune detection device based on micro-fluidic chip and using method |
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| CN201910313140.7A CN109946460B (en) | 2019-04-18 | 2019-04-18 | Chemiluminescence immune detection device based on micro-fluidic chip and using method |
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| CN109946460A CN109946460A (en) | 2019-06-28 |
| CN109946460B true CN109946460B (en) | 2023-09-22 |
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| CN113009136B (en) * | 2020-08-21 | 2024-04-05 | 东莞东阳光医疗智能器件研发有限公司 | Small multi-index detection sample analysis device |
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