CN111729696A - Microfluid detection chip adopting immune electrode - Google Patents
Microfluid detection chip adopting immune electrode Download PDFInfo
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- CN111729696A CN111729696A CN201911286651.0A CN201911286651A CN111729696A CN 111729696 A CN111729696 A CN 111729696A CN 201911286651 A CN201911286651 A CN 201911286651A CN 111729696 A CN111729696 A CN 111729696A
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Abstract
The invention discloses a microfluid detection chip adopting an immune electrode, which sequentially comprises a lower chip, a middle chip and an upper chip from bottom to top; the lower chip, the middle chip and the upper chip are matched to define a closed micro-channel and a plurality of mutually independent chambers; the micro-channel and the cavity are arranged on the middle chip in a penetrating way; the upper chip is provided with a sample inlet which is communicated with the cavity through the micro-channel; the microfluid detection chip further comprises an immune electrode system, wherein the immune electrode system comprises a standard electrode and a working electrode, the standard electrode comprises a gold-plated base layer, the working electrode comprises a gold-plated base layer, a conductive polymer layer and an antibody layer, and the gold-plated base layer, the conductive polymer layer and the antibody layer are sequentially attached from bottom to top. The chip has simple structure and convenient operation, improves the detection efficiency and precision and greatly reduces the consumption of resources.
Description
Technical Field
The invention belongs to the technical field of medical equipment, and particularly relates to a microfluid detection chip adopting an immune electrode.
Background
Microfluidics is a technology applied across various disciplines including engineering, physics, chemistry, microtechnology, and biotechnology. Microfluidics involves the study of minute quantities of fluids and how to manipulate, control and use such small quantities of fluids in various microfluidic systems and devices, such as microfluidic detection chips. For example: microfluidic biochips (known as "lab-on-a-chip") are used in the field of molecular biology to integrate assay operations for purposes such as analyzing enzymes and DNA, detecting biochemical toxins and pathogens, diagnosing diseases, and the like.
Microfluidic chip (microfluidic chip) is a hot spot area for the development of current micro total Analysis Systems (miniaturedtotal Analysis Systems). The micro-fluid detection chip analysis takes a chip as an operation platform, simultaneously takes analytical chemistry as a basis, takes a micro-electromechanical processing technology as a support, takes a micro-pipeline network as a structural characteristic, takes life science as a main application object at present, and is the key point of the development of the field of the current micro total analysis system. Its goal is to integrate the functions of the entire laboratory, including sampling, dilution, reagent addition, reaction, separation, detection, etc., on a microchip. The microfluid detection chip is a main platform for realizing microfluid technology. The device is characterized mainly by the fact that its active structures (channels, chambers and other functional elements) containing the fluid are of micron-scale dimensions at least in one dimension. Due to the micro-scale structure, the fluid exhibits and develops specific properties therein that differ from those of the macro-scale. Thus developing unique assay-generated properties. The characteristics and development advantages of the microfluid detection chip are as follows: the microfluid detection chip has the characteristics of controllable liquid flow, extremely less consumption of samples and reagents, ten-fold or hundred-fold improvement of analysis speed and the like, can simultaneously analyze hundreds of samples in a few minutes or even shorter time, and can realize the whole processes of pretreatment and analysis of the samples on line. The application of the micro total analysis system aims to realize the ultimate goal of the micro total analysis system, namely a lab-on-a-chip, and the key application field of the current work development is the field of life science.
In recent years, electrochemical immunosensors have attracted attention and are widely used for detecting tumor markers. When the immunosensor is used for continuous target detection, the reproducibility of the biomolecule probe is a big problem which restricts the practical application of the type of sensor. Therefore, the development of the immunosensor which can be produced in a large scale and has low cost can overcome the defects of complex operation, time and labor waste and the like caused by the fact that the biomolecular probe needs to be regenerated when the immunosensor is used as a disposable sensor. In the face of more and more special detection environments, development of new sensor technologies based on new materials and new processes has become a development trend. The flexible sensor based on the flexible matrix material has the characteristics of flexibility, extensibility, free bending or even folding, portability, wearability and the like, has flexible and various structural forms, can be randomly placed according to the requirements of measurement conditions, and is suitable for a plurality of special application scenes and environments.
Therefore, there is a need to develop a microfluidic chip suitable for an immuno-electrode, which can realize electrochemical detection, and has high detection sensitivity and stability, small interference between electrodes, and high accuracy.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a microfluid detection chip suitable for an immune electrode, which can realize electrochemical detection, and has high detection sensitivity and stability, small interference between electrodes and high accuracy.
In order to solve the technical problems, the technical scheme adopted by the invention is that the microfluid detection chip adopting the immune electrode sequentially comprises a lower chip, a middle chip and an upper chip from bottom to top; the lower chip, the middle chip and the upper chip are matched to define a closed micro-channel and a plurality of mutually independent chambers; the micro-channel and the cavity are arranged on the middle chip in a penetrating way; the upper chip is provided with a sample inlet which is communicated with the cavity through the micro-channel; the microfluid detection chip further comprises an immune electrode system, wherein the immune electrode system comprises a standard electrode and a working electrode, the standard electrode comprises a gold-plated base layer, the working electrode comprises a gold-plated base layer, a conductive polymer layer and an antibody layer, and the gold-plated base layer, the conductive polymer layer and the antibody layer are sequentially attached from bottom to top.
By adopting the technical scheme, the microfluid detection chip has the characteristics of high precision, high speed and low detection cost, is suitable for detection in an accurate medical link, adopts the chips with the structures of the lower chip, the middle chip and the upper chip, has reasonable design and simple and compact structure, and reduces the production cost; the detection reagent is embedded in the cavity in advance, the chip has a simple structure, is convenient to operate, improves the detection efficiency, and greatly reduces the consumption of resources; the rapid detection is realized, and the cost is reduced; the immune electrode system adopts a three-electrode system, a gold-plated base layer is a composite electrode of a standard electrode and a counter electrode, and an antibody/polypyrrole/gold-plated base layer is a detection electrode; then, carrying out immunoreaction on the PS microsphere-labeled antibody prepared based on the antigen-antibody specific reaction, an immunological electrode and a sample antigen to obtain a detection result of a sample index, so that the detection aim is realized by detecting the change of the dielectric constant in an immunological electrode system before and after the reaction; the polypyrrole has excellent conductivity, can accelerate electron transfer on the surface of the electrode, and improves the sensitivity of the immune electrode.
As a preferred technical scheme of the present invention, the chamber includes a reaction chamber and a waste liquid chamber, the reaction chamber and the waste liquid chamber are both disposed on the middle chip, the micro flow channel includes a first flow channel, a second cut-off valve site is disposed on the first flow channel between the reaction chamber and the waste liquid chamber, and the immunity electrode is divided into an upper immunity electrode disposed on the back of the upper chip and a lower immunity electrode disposed on the front of the lower chip; a gap is arranged between the upper layer immunity electrode and the lower layer immunity electrode; the upper layer immunity electrode is arranged on the back surface of the upper layer chip and the lower layer immunity electrode is arranged on the front surface of the lower layer chip and corresponds to the position of the reaction cavity of the middle layer chip, and the upper layer immunity electrode and the lower layer immunity electrode are communicated through the reaction cavity; the upper layer immunity electrode is a standard electrode, and the lower layer immunity electrode is a working electrode. The reaction cavity is internally embedded with a detection reagent in advance, and after blood to be detected flows into the reaction cavity, the upper layer immunity electrode and the lower layer immunity electrode are both contacted with the blood in the reaction cavity, so that the blood is in conductive communication; by designing the upper immune electrode and the lower immune electrode and arranging the electrodes of the microfluid detection chip adopting the immune electrodes in two layers, the mutual interference between the electrodes can be reduced, and the accuracy of a detection result is improved.
As a preferable technical scheme of the invention, the reaction cavity comprises a first reaction cavity and a second reaction cavity, the micro flow channel further comprises a second flow channel, a first cut-off valve position is arranged on the second flow channel between the first reaction cavity and the second reaction cavity, the upper layer immunity electrode is arranged on the back surface of the upper layer chip and the lower layer immunity electrode is arranged on the front surface of the lower layer chip and corresponds to the position of the second reaction cavity of the middle layer chip, and the upper layer immunity electrode and the lower layer immunity electrode are communicated through the second reaction cavity. The first cut-off valve site is arranged to ensure that when the first reaction cavity is used for reaction, the blood sample and the reacted reagent are only in the first reaction cavity and cannot flow into other cavities, so that the first reaction cavity can be used for reaction independently.
As a preferable technical scheme of the present invention, the chamber further includes a cleaning liquid chamber, the microchannel further includes a sub-channel, the second channel is provided with the sub-channel extending outward from the outside of the first channel between the second reaction chamber and the first stop valve, and the sub-channel is connected to the cleaning liquid chamber. The second interception valve site is arranged to ensure that when the second reaction cavity reacts, the blood sample and the reacted reagent are only in the second reaction cavity and cannot be left in the waste liquid cavity, and meanwhile, when the reaction is ensured, waste liquid in the waste liquid cavity cannot pollute the second reaction cavity, and the reaction of the second reaction cavity is prevented from being interfered. The cleaning liquid adopts the liquid bag to set up in the cleaning liquid chamber, and during the use, supporting detecting instrument's power rod device presses the liquid bag, and liquid bag front end and subchannel intercommunication department are broken by the extrusion to the inside liquid of liquid bag flows out.
As a preferable technical scheme of the invention, the reaction cavity II is communicated with the waste liquid cavity through the first flow channel.
As a preferred technical scheme of the invention, an upper layer connecting through hole is arranged on the upper layer chip, a lower layer connecting through hole is arranged on the lower layer chip, and the upper layer immunity electrode is in contact connection with a matched detection instrument through the lower layer connecting through hole; and the lower layer immunity electrode is in contact connection with the matched detection instrument through the upper layer connecting through hole. The arrangement ensures that the immunity electrode can be connected with a matched instrument through the upper-layer communicating hole and the lower-layer communicating hole without additionally arranging a connecting end with the matched instrument.
As a preferred technical scheme of the invention, the middle layer chip is provided with a liquid receiving port, the position of the liquid receiving port corresponds to that of the sample inlet, the micro flow channel further comprises a third flow channel, and the first reaction cavity is communicated with the liquid receiving port through the third flow channel.
As a preferable technical scheme of the invention, the chamber further comprises a buffer liquid cavity, the sub-runners extend outwards along the direction outside the sub-runners and are provided with a first sub-runner, and the first sub-runner is connected with the buffer liquid cavity. The buffer solution adopts the liquid bag setting in the buffer solution chamber, and during the use, supporting detecting instrument's power rod device presses the liquid bag, and liquid bag front end and subchannel intercommunication department are broken by the extrusion to the inside liquid of liquid bag flows out.
As a preferred technical scheme of the present invention, the upper chip is further provided with at least one vent hole, and the vent hole is arranged at one end of the upper chip and at a position corresponding to the waste liquid cavity. The upper-layer chip is provided with at least one vent hole in a penetrating manner, so that the flow resistance of the fluid to be tested is reduced, the flow is quicker, and the chamber is quickly filled; the exhaust hole is arranged to facilitate the flow of the sample, the sample introduction is convenient, if the exhaust hole is not arranged, the sample cannot flow into the reaction cavity to react, and the detection reagent is embedded in the reaction cavity in advance.
As a preferred technical scheme of the invention, the sample inlet is provided with a sample injection cover, and the sample injection cover covers the sample inlet after sample injection so as to enable a sample to flow.
As a preferred technical scheme of the invention, the gold-plated base layer comprises a base layer and a gold layer; the working electrode further includes a porous protective layer disposed on a surface of the antibody layer. Wherein the gold-plated base layer is a purchased finished product, and gold is connected with the base layer by means of vacuum magnetic sputtering, electroplating, silk-screen printing and the like; the base layer is a flexible base layer and is made of materials such as PET, PP, PE, ABS and the like; wherein, the PET has strong hardness and bending resistance, and the surface is smooth and can be coated; the surface of the flexible base layer is plated with gold, so that the base layer has conductivity, the gold nano layer or the non-gold nano layer has good conductivity, stronger biocompatibility and protein activity can be maintained to a certain extent, and the conductivity and the stability of the immune electrode are improved; wherein the porous protection layer is used for carrying out porous modification treatment on the surface of the antibody layer.
As a preferred technical scheme of the invention, the lower chip, the middle chip and the upper chip are bonded into a whole in a double-sided gluing mode through the middle chip.
As a preferred technical scheme of the invention, the middle layer chip is double-sided adhesive tape, the upper layer chip and/or the lower layer chip is made of any one of PMMA, PP, PE and PET, and hydrophilic films are arranged on the surfaces of the upper layer chip and the lower layer chip, so that a sample quickly flows into the micro-channel through the sample inlet and then flows into each chamber. The pressure-sensitive adhesive tape is preferably selected as the middle chip, and the technical scheme is adopted, so that the material is easy to obtain, and the thickness of the pressure-sensitive adhesive tape can be precisely controlled by the manufacturing process of the pressure-sensitive adhesive tape, so that the depth and the size of the micro-channel can be precisely controlled, and the depth of the cavity can be conveniently controlled, so that the thickness deviation of each cavity of the micro-fluid detection chip is small, the consistency is high, and the detection accuracy is improved; the surfaces of the upper chip and the lower chip are provided with hydrophilic films, so that samples can flow into the micro-channel and each cavity through the sample inlet more quickly, the flow speed is accelerated, and the detection efficiency can be improved.
As the preferred technical scheme of the invention, the thickness of the middle layer chip is 0.1-1.0 mm; the surface of the lower chip is flat, the depth of a closed micro-channel defined by the lower chip, the middle chip and the upper chip in a matched mode is 0.1-1.0 mm, and the width of the cavity defined by the lower chip, the middle chip and the upper chip in a matched mode is 1.0-2.0 mm.
Compared with the prior art, the microfluid detection chip adopting the immune electrode improves the accuracy of the detection result; meanwhile, a reaction chamber, a waste liquid chamber, a cleaning liquid chamber and a buffer liquid chamber in a specific structural form are designed, so that detection reaction is finished in a microfluid detection chip, and a reaction result is obtained; the chip has simple structure and convenient operation, improves the detection efficiency and precision and greatly reduces the consumption of resources; and the rapid detection is realized, and the cost is reduced.
Drawings
The following further detailed description of embodiments of the invention is made with reference to the accompanying drawings:
FIG. 1 is a schematic perspective view of the entire front face of a microfluidic detection chip using an immuno-electrode according to the present invention;
FIG. 2 is a schematic diagram of a three-layer explosion structure of a microfluidic detection chip using an immuno-electrode according to the present invention;
FIG. 3 is a schematic diagram of the front structure of the upper chip of the microfluidic detection chip using the immuno-electrode according to the present invention;
FIG. 4 is a schematic diagram of the back side structure of the upper chip of the microfluidic detection chip using the immuno-electrode according to the present invention;
FIG. 5 is a schematic diagram of the front side of a middle chip of the microfluidic detection chip using an immuno-electrode according to the present invention;
FIG. 6 is a schematic diagram showing the reverse structure of a middle chip of the microfluidic detection chip using an immuno-electrode according to the present invention;
FIG. 7 is a schematic diagram of the structure of the front surface of the lower chip of the microfluidic detection chip using the immuno-electrode according to the present invention;
FIG. 8 is a schematic diagram showing the structure of the lower chip of the microfluidic detection chip using an immuno-electrode according to the present invention;
FIG. 9 is a block diagram of an immunizing electrode of the present invention;
wherein: 1-lower chip; 101-lower layer communicating holes; 2-middle layer chip; 201-middle layer communication hole; 202-a liquid receiving port; 3-upper chip; 301-upper layer connecting through hole; 4-upper immuno-electrode; 5-lower immunization electrode; 6-micro flow channel; 601-runner one; 602-channel two; 603-runner III; 604-a shunt; 605-branch channel one; 7-air vent; 8-a sample inlet; 901-a first reaction chamber; 902-reaction chamber two; 903-waste liquid cavity; 904-buffer chamber; 905-cleaning liquid cavity; 10-a base layer; 11-gold layer; 12-a conductive polymer layer; 13-an antibody layer; 14-porous protective layer.
Detailed Description
Example 1: as shown in fig. 1 to 9, the micro-fluidic detection chip using the immuno-electrode sequentially comprises a lower chip 1, a middle chip 2 and an upper chip 3 from bottom to top; the lower chip 1, the middle chip 2 and the upper chip 3 are matched to define a closed micro-channel and a plurality of mutually independent chambers; the micro flow channel 6 and the cavity are both arranged on the middle layer chip 2 in a penetrating way; the upper chip 3 is provided with a sample inlet 8, and the sample inlet 8 is communicated with the cavity through the micro-channel 6; the microfluid detection chip further comprises an immune electrode system, wherein the immune electrode system comprises a standard electrode and a working electrode, the standard electrode comprises a gold-plated base layer, the working electrode comprises a gold-plated base layer, a conductive polymer layer 12 and an antibody layer 13, the gold-plated base layer comprises a base layer 10 and a gold layer 11, the base layer 10, the gold layer 11, the conductive polymer layer 12 and the antibody layer 13 are sequentially attached from bottom to top, and the base layer 10 is a PET flexible base layer; the cavity comprises a reaction cavity and a waste liquid cavity 903, the reaction cavity and the waste liquid cavity 903 are both arranged on the middle chip 2, the micro flow channel 6 comprises a first flow channel 601, a second cut valve position point is arranged on the first flow channel 601 between the reaction cavity and the waste liquid cavity 903, and the immune electrode is divided into an upper immune electrode 4 arranged on the back surface of the upper chip 3 and a lower immune electrode 5 arranged on the back surface of the lower chip 1; a gap is arranged between the upper layer immunity electrode 4 and the lower layer immunity electrode 5; the upper layer immunity electrode 4 is arranged on the back surface of the upper layer chip 3 and the lower layer immunity electrode 5 is arranged on the front surface of the lower layer chip 1 and is corresponding to the position of the reaction cavity of the middle layer chip 2, and the upper layer immunity electrode 4 is communicated with the lower layer immunity electrode 5 through the reaction cavity; the upper layer immunity electrode 4 is a standard electrode, and the lower layer immunity electrode 5 is a working electrode; the detection reagent is embedded in the reaction cavity in advance, and after blood to be detected flows into the reaction cavity, the upper layer immunity electrode 4 and the lower layer immunity electrode 5 are both contacted with the blood in the reaction cavity, so that the blood is in conductive communication; the reaction cavity comprises a first reaction cavity 901 and a second reaction cavity 902, the micro flow channel further comprises a second flow channel 602, a first cut-off valve position is arranged on the second flow channel 602 between the first reaction cavity 901 and the second reaction cavity 902, the upper layer immunity electrode 4 is arranged on the back surface of the upper layer chip 3 and the lower layer immunity electrode 5 is arranged on the front surface of the lower layer chip 1 and corresponds to the position of the second reaction cavity 902 of the middle layer chip 2, and the upper layer immunity electrode 4 is communicated with the lower layer immunity electrode 5 through the second reaction cavity 902; the chamber further comprises a cleaning liquid cavity 905, the micro flow channel 6 further comprises a sub flow channel 604, the second flow channel 602 extends outwards from the position between the second reaction cavity 902 and the first stop valve position towards the outer side of the second flow channel 602, and the sub flow channel 604 is connected with the cleaning liquid cavity 905; the second reaction cavity 902 is communicated with the waste liquid cavity 903 through the first flow channel 601; the upper chip 3 is provided with an upper layer communicating hole 301, the lower chip 1 is provided with a lower layer communicating hole 101, and the upper layer immunity electrode 4 is in contact connection with a matched detection instrument through the lower layer communicating hole 101; the lower layer immunity electrode 5 is in contact connection with the matched detection instrument through the upper layer connection through hole 301; the middle chip 2 is provided with a liquid receiving port 202, the position of the liquid receiving port 202 corresponds to that of the sample inlet 8, the micro flow channel 6 further comprises a third flow channel 603, and the first reaction cavity 901 is communicated with the liquid receiving port 202 through the third flow channel 603; the chamber further comprises a buffer liquid cavity 904, the sub-runner 604 extends outwards along the direction outside the sub-runner 604 to form a first sub-runner 605, and the first sub-runner 605 is connected with the buffer liquid cavity 904; the upper chip 3 is further provided with at least one vent hole 7, and the vent hole 7 is arranged at one end of the upper chip 3 and at a position corresponding to the waste liquid cavity 903; the arrangement of the exhaust hole 7 is beneficial to the flow of the sample and is convenient for sample introduction; a sample injection cover is arranged on the sample injection port 8, and the sample injection cover covers the sample injection port 8 after sample injection so as to enable a sample to flow; the immunity electrode also comprises a porous protection layer 14, and the porous protection layer 14 is prepared on the surface of the antibody layer 13.
Example 2: the microfluid detection chip adopting the immune electrode sequentially comprises a lower chip 1, a middle chip 2 and an upper chip 3 from bottom to top; the lower chip 1, the middle chip 2 and the upper chip 3 are matched to define a closed micro-channel and a plurality of mutually independent chambers; the micro flow channel 6 and the cavity are both arranged on the middle layer chip 2 in a penetrating way; the upper chip 3 is provided with a sample inlet 8, and the sample inlet 8 is communicated with the cavity through the micro-channel 6; the microfluid detection chip further comprises an immunity electrode, wherein the immunity electrode comprises a standard electrode and a working electrode, the standard electrode comprises a gold-plated base layer, the working electrode comprises a gold-plated base layer, a conductive polymer layer 12, an antibody layer 13 and a porous protection layer 14, the gold-plated base layer comprises a base layer 10 and a gold layer 11, the base layer 10, the gold layer 11, the conductive polymer layer 12, the antibody layer 13 and the porous protection layer 14 are sequentially attached from bottom to top, and the base layer 10 is a PET flexible base layer; the cavity comprises a reaction cavity and a waste liquid cavity 903, the reaction cavity and the waste liquid cavity 903 are both arranged on the middle chip 2, the micro flow channel 6 comprises a first flow channel 601, a second cut valve position point is arranged on the first flow channel 601 between the reaction cavity and the waste liquid cavity 903, and the immune electrode is divided into an upper immune electrode 4 arranged on the back surface of the upper chip 3 and a lower immune electrode 5 arranged on the back surface of the lower chip 1; a gap is arranged between the upper layer immunity electrode 4 and the lower layer immunity electrode 5; the upper layer immunity electrode 4 is arranged on the back surface of the upper layer chip 3 and the lower layer immunity electrode 5 is arranged on the front surface of the lower layer chip 1 and is corresponding to the position of the reaction cavity of the middle layer chip 2, and the upper layer immunity electrode 4 is communicated with the lower layer immunity electrode 5 through the reaction cavity; the upper layer immunity electrode 4 is a standard electrode, and the lower layer immunity electrode 5 is a working electrode; the detection reagent is embedded in the reaction cavity in advance, and after blood to be detected flows into the reaction cavity, the upper layer immunity electrode 4 and the lower layer immunity electrode 5 are both contacted with the blood in the reaction cavity, so that the blood is in conductive communication; the reaction cavity comprises a first reaction cavity 901 and a second reaction cavity 902, the micro flow channel further comprises a second flow channel 602, a first cut-off valve position is arranged on the second flow channel 602 between the first reaction cavity 901 and the second reaction cavity 902, the upper layer immunity electrode 4 is arranged on the back surface of the upper layer chip 3 and the lower layer immunity electrode 5 is arranged on the front surface of the lower layer chip 1 and corresponds to the position of the second reaction cavity 902 of the middle layer chip 2, and the upper layer immunity electrode 4 is communicated with the lower layer immunity electrode 5 through the second reaction cavity 902; the chamber further comprises a cleaning liquid cavity 905, the micro flow channel 6 further comprises a sub flow channel 604, the second flow channel 602 extends outwards from the position between the second reaction cavity 902 and the first stop valve position towards the outer side of the second flow channel 602, and the sub flow channel 604 is connected with the cleaning liquid cavity 905; the second reaction cavity 902 is communicated with the waste liquid cavity 903 through the first flow channel 601; the upper chip 3 is provided with an upper layer communicating hole 301, the lower chip 1 is provided with a lower layer communicating hole 101, and the upper layer immunity electrode 4 is in contact connection with a matched detection instrument through the lower layer communicating hole 101; the lower layer immunity electrode 5 is in contact connection with the matched detection instrument through the upper layer connection through hole 301; the middle chip 2 is provided with a liquid receiving port 202, the position of the liquid receiving port 202 corresponds to that of the sample inlet 8, the micro flow channel 6 further comprises a third flow channel 603, and the first reaction cavity 901 is communicated with the liquid receiving port 202 through the third flow channel 603; the chamber further comprises a buffer liquid cavity 904, the sub-runner 604 extends outwards along the direction outside the sub-runner 604 to form a first sub-runner 605, and the first sub-runner 605 is connected with the buffer liquid cavity 904; the upper chip 3 is further provided with at least one vent hole 7, and the vent hole 7 is arranged at one end of the upper chip 3 and at a position corresponding to the waste liquid cavity 903; the arrangement of the exhaust hole 7 is beneficial to the flow of the sample and is convenient for sample introduction; a sample injection cover is arranged on the sample injection port 8, and the sample injection cover covers the sample injection port 8 after sample injection so as to enable a sample to flow; the immunity electrode also comprises a porous protection layer 14, and the porous protection layer 14 is prepared on the surface of the antibody layer 13; the lower chip 1, the middle chip 2 and the upper chip 3 are bonded into a whole in a double-sided gluing mode through the middle chip 2; the middle chip 2 is a double-sided adhesive tape, the upper chip 3 and/or the lower chip 1 is made of any one of PMMA, PP, PE and PET, and the surfaces of the upper chip 3 and the lower chip 1 are provided with hydrophilic films, so that a sample can rapidly flow into the micro-channel 6 through the sample inlet 8 and then flow into each chamber. The pressure-sensitive adhesive tape is preferably selected as the middle chip 2, and the technical scheme is adopted, so that the material is easy to obtain, and the thickness of the pressure-sensitive adhesive tape can be precisely controlled by the manufacturing process of the pressure-sensitive adhesive tape, so that the depth and the size of the micro-channel can be precisely controlled by adopting the technical scheme, and the depth of the cavity can be conveniently controlled, so that the thickness deviation of each cavity of the micro-fluid detection chip is small, the consistency is high, and the detection accuracy is improved; hydrophilic films are arranged on the surfaces of the upper chip 3 and the lower chip 1; the thickness of the middle layer chip 2 is 0.1-1.0 mm; the surface of the lower-layer chip 1 is flat, the depth of a closed micro-channel 6 defined by the lower-layer chip 1, the middle-layer chip 2 and the upper-layer chip 3 in a matched mode is 0.1-1.0 mm, and the width of a cavity defined by the lower-layer chip 1, the middle-layer chip 2 and the upper-layer chip in a matched mode is 1.0-2.0 mm.
When in specific use: firstly, closing the first interception valve site, enabling a buffer liquid sac in the buffer liquid cavity 904 to be broken under the action of a power rod device matched with a detection instrument, enabling a buffer liquid in the liquid sac to enter the second reaction cavity 902 under the action of liquid driving by a power rod, then closing the second interception valve site, reading data on the detection instrument within 30 seconds, and opening the second interception valve site to enable liquid in the second reaction cavity 902 to enter the waste liquid cavity 903; injecting a blood sample into the sample inlet 8, covering a sample injection cover, enabling the sample to flow from the liquid receiving port 202 to the first reaction cavity 901 through the third flow channel 603, reacting with the internal fixed antibody for 1-5 minutes, opening the first interception valve site after the reaction is finished, enabling the sample to enter the second reaction cavity 902, closing the first interception valve site, ensuring that the second interception valve site is in a closed state, reacting for 1-5 minutes, and opening the second interception valve site; the effect of the power rod device of the matched detection instrument enables the cleaning liquid bag in the cleaning liquid cavity 905 to be broken, the internal cleaning liquid enters the second reaction cavity 902 to clean the immune electrode, the cleaning waste liquid enters the waste liquid cavity, the instrument receives an electric signal in the reaction cavity, and the instrument software calculates to obtain the index content of the sample.
The foregoing illustrates and describes the principles, general features, and advantages of the present invention. It will be understood by those skilled in the art that the present invention is not limited to the above embodiments, and the above embodiments and descriptions are only illustrative of the principles of the present invention, and that various changes and modifications may be made without departing from the spirit and scope of the present invention, such as the layout structure of the chambers, the arrangement of the immuno-electrodes (e.g., the immuno-electrodes may also be arranged on the same chip), and the shape of the immuno-electrodes, and other minor modifications may be made, and such changes and modifications are within the scope of the present invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (11)
1. A microfluid detection chip adopting an immune electrode comprises a chip body, wherein the chip body sequentially comprises a lower chip, a middle chip and an upper chip from bottom to top; the lower chip, the middle chip and the upper chip are matched to define a closed micro-channel and a plurality of mutually independent chambers; the micro-channel and the cavity are arranged on the middle chip in a penetrating way; the upper chip is provided with a sample inlet which is communicated with the cavity through the micro-channel; the microfluid detection chip is characterized by further comprising an immune electrode system, wherein the immune electrode system comprises a standard electrode and a working electrode, the standard electrode comprises a gold-plated base layer, the working electrode comprises a gold-plated base layer, a conductive polymer layer and an antibody layer, and the gold-plated base layer, the conductive polymer layer and the antibody layer are sequentially attached from bottom to top.
2. The microfluidic detection chip adopting the immuno-electrode according to claim 1, wherein the chamber comprises a reaction chamber and a waste liquid chamber, the reaction chamber and the waste liquid chamber are both disposed on the middle chip, the microchannel comprises a first flow channel, a second shut-off valve site is disposed on the first flow channel between the reaction chamber and the waste liquid chamber, and the immuno-electrode is divided into an upper immuno-electrode disposed on the back surface of the upper chip and a lower immuno-electrode disposed on the front surface of the lower chip; a gap is arranged between the upper layer immunity electrode and the lower layer immunity electrode; the upper layer immunity electrode is arranged on the back surface of the upper layer chip and the front surface of the lower layer chip are arranged corresponding to the position of the reaction cavity of the middle layer chip, the upper layer immunity electrode is communicated with the lower layer immunity electrode through the reaction cavity, the upper layer immunity electrode is a standard electrode, and the lower layer immunity electrode is a working electrode.
3. The microfluidic detection chip adopting the immunity electrode as claimed in claim 2, wherein the reaction chamber comprises a first reaction chamber and a second reaction chamber, the microchannel further comprises a second flow channel, the first flow channel between the first reaction chamber and the second reaction chamber is provided with a first cut-off valve site, the upper immunity electrode is disposed on the back surface of the upper chip and the lower immunity electrode is disposed on the front surface of the lower chip in a position corresponding to the second reaction chamber of the middle chip, and the upper immunity electrode and the lower immunity electrode are communicated through the second reaction chamber.
4. The microfluidic detection chip using the immuno-electrode according to claim 3, wherein the chamber further comprises a cleaning solution chamber, the microchannel further comprises a bypass, the bypass is provided between the second reaction chamber and the first stop valve and extends outwards from the second reaction chamber to the outside of the microchannel, and the bypass is connected to the cleaning solution chamber.
5. The microfluidic detection chip using an immuno electrode of claim 3, wherein the reaction chamber two-way channel is connected to the waste liquid chamber through the first flow channel.
6. The microfluidic detection chip adopting the immuno-electrode according to claim 4, wherein the upper chip is provided with an upper layer communication hole, the lower chip is provided with a lower layer communication hole, and the upper layer immuno-electrode is in contact connection with a matched detection instrument through the lower layer communication hole; and the lower layer immunity electrode is in contact connection with the matched detection instrument through the upper layer connecting through hole.
7. The microfluidic chip for detecting micro-fluid using immuno-electrodes of claim 4, wherein the middle chip has a liquid receiving port corresponding to the position of the sample inlet, the micro-channel further comprises a third channel, and the first reaction chamber is communicated with the liquid receiving port through the third channel.
8. The microfluid detection chip adopting an immuno electrode according to claim 4, wherein the chamber further comprises a buffer liquid cavity, the first sub-channel extends outwards along the direction outside the first sub-channel, and the first sub-channel is connected with the buffer liquid cavity.
9. The microfluidic detection chip adopting an immuno-electrode according to claim 4, wherein the upper chip is further provided with at least one vent hole, and the vent hole is arranged at one end of the upper chip and at a position corresponding to the waste liquid chamber.
10. The microfluidic detection chip using the immuno electrode of claim 7, wherein a sample injection cover is disposed on the sample injection port, and the sample injection cover covers the sample injection port after sample injection to allow the sample to flow.
11. The microfluidic detection chip using an immuno electrode of claim 1, wherein said gold-plated base layer comprises a base layer and a gold layer; the working electrode further includes a porous protective layer disposed on a surface of the antibody layer.
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| CN201911286651.0A CN111729696A (en) | 2019-12-14 | 2019-12-14 | Microfluid detection chip adopting immune electrode |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021115491A1 (en) * | 2019-12-14 | 2021-06-17 | 南京岚煜生物科技有限公司 | Preparation method for immunoelectrode |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021115491A1 (en) * | 2019-12-14 | 2021-06-17 | 南京岚煜生物科技有限公司 | Preparation method for immunoelectrode |
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