CN116212990A - Array electrode card with multiple microelectrodes integrated for multi-parameter biochemical molecular detection and microfluidic detection card - Google Patents
Array electrode card with multiple microelectrodes integrated for multi-parameter biochemical molecular detection and microfluidic detection card Download PDFInfo
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Abstract
The invention provides a microfluidic detection card for multi-parameter biochemical molecule detection by using an array electrode card with a plurality of microelectrodes integrated, which comprises a detection card main body, a microfluidic system, an array electrode mounting clamping groove and an electrode card, wherein a plurality of microelectrodes are integrated on the surface of the electrode card; the microfluidic system comprises a sample inlet, a sample pipeline system, a calibration liquid system, an electrode assembly area and a liquid guiding-out system; the electrode assembly area is provided with an array electrode mounting clamping groove, a single electrode card can be loaded in the single clamping groove, a plurality of microelectrodes are integrated on the front surface of the single electrode card, the back surface of the electrode card is provided with the same number of electrode pins, and the microelectrodes and the electrode pins are correspondingly connected with the metal circuit one by one through conducting sites on the surface of the electrode sheet. The invention can reduce the cost and processing difficulty of the chip, improve the detection speed, sensitivity and accuracy while ensuring the simultaneous detection of multiple parameters, and realize the simultaneous detection of multiple indexes in the biochemical electrolyte.
Description
Technical Field
The invention belongs to the field of biomedical sensors, and relates to a microfluidic detection card with an array electrode card integrated by a plurality of microelectrodes for multi-parameter biochemical molecular detection.
Background
The current most novel POCT chemical sensing instrument product can realize that a plurality of samples are detected simultaneously, so that the detection time is greatly saved, and meanwhile, the instant detection is faster. POCT devices are mostly realized by using a biosensor, and the biological enzyme molecules are immobilized on a solid phase interface of a micro-analysis device, and after the analyte is specifically identified, the detection is performed by using an electrochemical or optical method, and the reading is immediately given. Typical biosensor-based commercial POCT devices are represented by blood glucose meters, blood gas analyzers, and are mainly used for blood glucose monitoring, blood gas and electrolyte analysis.
Aiming at a biochemical detection consumable device for simultaneously detecting a plurality of material indexes at one time, related technology exists at present, and specifically, microelectrodes for detecting specific biochemical molecules are integrated on an integral detection device and are applied to multi-parameter electrochemical detection. At present, the technology for combining a microfluidic system with POCT products exists at home and abroad. The microfluidic is a technology for controlling and operating a microfluidic on a chip with a micro-pipeline, and the conventional POCT related microfluidic system has defects such as solidification and blockage of liquid to be detected in the micro-fluidic pipeline, easy backflow and miscibility of the liquid to be detected, low liquid flow rate, low detection efficiency, influence of waste gas generated by reaction and the like.
In view of the above, the present invention aims to provide a microfluidic detection card with a plurality of microelectrode integrated array electrode cards for multi-parameter biochemical molecular detection, which integrates the functions of preventing solidification and blockage of a liquid to be detected, mixing of a calibration solution, detection of a sample solution, recovery of waste liquid, discharge of waste gas, etc. on one chip, reduces the cost and processing difficulty of the chip, ensures simultaneous detection of multiple parameters, improves the detection speed, sensitivity and accuracy, and realizes simultaneous detection of multiple indexes in biochemical electrolytes.
Disclosure of Invention
In summary, the present invention provides a microfluidic detection card with an array electrode card integrated by a plurality of microelectrodes for multi-parameter biochemical molecular detection.
The invention aims to provide an array electrode card with a plurality of microelectrodes for multi-parameter biochemical molecular detection, which comprises a detection card main body, a microfluidic system arranged in the detection card main body, an array electrode mounting clamping groove and an electrode card loaded in the array electrode mounting clamping groove, wherein a plurality of microelectrodes are integrated on the surface of the electrode card; the microfluidic system comprises a sample inlet, a sample pipeline system, a calibration liquid system, an electrode assembly area and a liquid guiding-out system; the electrode assembly area is provided with an array electrode mounting clamping groove, the array electrode mounting clamping groove is formed by arranging a plurality of electrode mounting clamping grooves in an array manner, a piece of electrode card can be loaded in a single clamping groove, a plurality of microelectrodes are integrated on the front surface of the single electrode card, the back surface of the electrode card is provided with the same number of electrode pins, and the microelectrodes and the electrode pins are correspondingly connected with metal circuits one by one through conducting sites on the surface of an electrode sheet; a sample test groove is formed in each clamping groove of the array electrode mounting clamping groove, the front surface of the electrode card is downwards loaded in the clamping groove, and microelectrodes modified on the surface of the electrode card are correspondingly embedded into the sample test grooves; one end of the electrode assembly area is connected with the sample injection pipeline system, and the other end of the electrode assembly area is communicated with the liquid guiding-out system; the sample introduction pipe system passes through the electrode assembly area, specifically each clamping groove of the array electrode installation clamping groove, and is connected with the sample test groove.
Further, be provided with the exhaust hole on the detection card main part, exhaust hole and liquid export system intercommunication.
Further, the detection card main body is also provided with a detection card positioning hole.
In the microfluidic system, the sample injection pipeline system comprises a pretreatment chamber, a pretreatment buffer channel, a miniature check valve and a sample injection liquid channel, wherein the sample injection liquid channel is connected with a sample injection port and is sequentially communicated with the pretreatment chamber, the pretreatment buffer channel and the miniature check valve, and the calibration liquid system is connected with the miniature check valve.
Furthermore, immune magnetic beads can be placed in the pretreatment chamber, and anticoagulant drugs can be placed in the pretreatment chamber or the pretreatment chamber is empty.
Further, the pretreatment buffer channel can be a micro flow channel formed by a plurality of micro saddle-shaped structures.
In the microfluidic system, the calibration liquid system comprises a calibration liquid channel and a calibration liquid package, and a flow guide valve is arranged between the calibration liquid channel and the calibration liquid package; the calibration fluid channel is connected to the sample introduction tubing.
In the microfluidic system, the liquid guiding-out system comprises a simple check valve, a waste liquid tank and a sample liquid outlet channel, wherein the sample liquid outlet channel is connected with a sample inlet pipeline system, and the sample liquid outlet channel is sequentially communicated with the simple check valve and the waste liquid tank.
Further, the simple check valve structure includes a tesla valve structure.
In the array electrode mounting clamping groove, the sample test groove is a groove with a fixed volume.
Further, each clamping groove of the array electrode mounting clamping groove is internally provided with a positioning column, the surface of the electrode card is provided with a positioning hole, the positioning column corresponds to the positioning hole in position and is in accordance with the size, and the electrode card can be fixed in the array electrode mounting clamping groove through the positioning hole.
The microelectrode integrated on the surface of the electrode card comprises a reference electrode, an auxiliary electrode and a working electrode, wherein the working electrode comprises a calibration electrode.
Further, the microelectrode with the surface modified by the electrode card comprises a reference electrode, an auxiliary electrode, a working electrode A, a working electrode B and a calibration electrode.
As shown in fig. 1, fig. 2 and fig. 3, the structure, front and back of the microfluidic detection card provided by the invention are respectively shown.
The sample inlet can inject blood/test sample into the microfluidic pipeline through a 1-3 ml needle tube; the pretreatment chamber is used for pretreatment of sample injection solution, such as separation, filtration, mixing and the like, and magnetic bead microspheres can be placed in the pretreatment chamber for molecular extraction of RNA and ctDNA, namely, the pretreatment chamber is mixed with a sample for DNA detection; the pretreatment buffer channel has the functions of uniformly mixing and reducing the flow rate, can improve the pretreatment efficiency of the sample solution and prevent insufficient reaction in the pretreatment chamber; the miniature check valve is of a miniature inverted truncated cone-shaped structure, the detection card is arranged in the vertical direction when being horizontally placed, the opening is large at the upper part and small at the lower part, and the sample is prevented from flowing backwards through the flowing resistance; the flow guide valve can guide the calibration liquid to the correct circulation direction and has a structure with a certain height difference to prevent the liquid from flowing backwards into the calibration liquid bag, particularly the calibration liquid bag has a higher direction and is lowered along the outflow direction of the calibration liquid; the calibration liquid package is used for loading calibration liquid with standard concentration, and the calibration liquid can flow out through a mechanical device or a manual extrusion mode.
The electrode assembly area digs out an area and is used for sample detection on detecting the card main part, for array electrode installation draw-in groove, specifically can divide into 6 draw-in grooves that the specification is unanimous, all is provided with the recess of fixed volume in every draw-in groove, as the sample test groove, is provided with the reference column in both corners department simultaneously, and the position of sample test groove and reference column corresponds with microelectrode detection site and locating hole on the electrode card respectively, and when the electrode card was installed in the draw-in groove inside, its integrated microelectrode was just fixed in the sample test groove.
As shown in fig. 4 and fig. 5, the front and back structures of the electrode card are respectively shown.
Five microelectrodes are integrated on the front face of the electrode card, and the microelectrode card comprises 1 reference electrode, 1 auxiliary electrode and 3 working electrodes, wherein the 1 working electrodes can be used as calibration electrodes. The surface of the reference electrode is modified with silver-silver chloride, the surface of the auxiliary electrode is modified with a platinum layer, the surface of the working electrode can be modified with a gold layer, and different sensitive substances can be modified according to detection indexes. Each electrode card can detect at least two material indexes simultaneously.
The calibration electrode can achieve the purpose of self-calibration, so that the detection result of the working electrode is more accurate.
The back of the electrode card is provided with metal pins with the same number as the microelectrodes, and the microelectrodes are correspondingly connected with the metal pins through metal circuits in the electrode card or metal circuits of the through holes.
The electrode card is installed in the array electrode installation card slot with the front face downwards, and the microelectrode is correspondingly embedded into the sample test slot.
The number of the clamping grooves of the array electrode mounting clamping groove can be changed according to the number of the detection indexes, meanwhile, the size of the clamping grooves and the size of the electrode cards can be changed according to requirements, and the number of the positioning columns can be changed according to the number and the size of the electrode cards.
The sample liquid channel of the detection card main body sequentially passes through six sample test grooves of the array electrode mounting clamping groove, and is connected with the sample liquid channel at the outlet, and the simple check valve adopts a special structure to prevent the liquid of the waste liquid groove from flowing backwards; the waste liquid tank can be loaded with tested liquid, including calibration liquid and test sample, and the exhaust hole is used for exhausting gas in the flow channel, so that the liquid can flow in the micro flow channel conveniently, and the waste liquid tank has a simple anti-overflow structure.
All the structures except the electrode card needing to be additionally arranged are all contained in the detection card main body, the processing can be finished at one time by an injection molding method, and additional parts are not needed to be additionally arranged, so that the processing steps of a microfluidic system are greatly simplified, the integration of the microfluidic and sensing detection card is realized, the electrode card is fixed in the detection card slot, various biochemical molecules are simultaneously detected through a microelectrode integration scheme, and the multi-parameter electrochemical detection of the microfluidic sensing detection card is realized.
Fig. 6 is a schematic diagram of liquid sample injection of the microfluidic detection card according to the present invention.
When the sample is added, the detection card is horizontally placed, the front face is upward, the liquid to be measured is injected from the sample inlet, flows through the pretreatment chamber, flows into the pretreatment buffer channel and then flows through the miniature check valve, meanwhile, the calibration liquid in the calibration liquid bag is extruded and flows out by using a mechanical device or manpower to extrude the calibration liquid bag, flows into the calibration liquid channel after passing through the flow guide valve, and is mixed with the liquid to be measured in the sample liquid channel through the miniature check valve; the mixed liquid sequentially passes through the sample liquid channel, the sample testing groove and the microelectrode which is contacted with the front surface of the electrode card for modification, then passes through the sample liquid channel, flows through the simple check valve, enters the waste liquid groove, generates gas in the process and is discharged through the exhaust hole connected with the waste liquid groove, and the detection electric signal is transmitted through the electrode pin on the back surface of the electrode card.
The beneficial effects of the invention are as follows:
(1) According to the invention, through the design of a microfluidic structure, the functions of preventing the to-be-detected liquid from solidifying and blocking pretreatment, mixing the calibration liquid, detecting the sample liquid, recovering the waste liquid, discharging the waste gas and the like are concentrated on one chip, so that the cost and the processing difficulty of the chip are reduced, the simultaneous detection of multiple parameters is ensured, the detection speed, the sensitivity and the accuracy are improved, and the simultaneous detection of multiple indexes in the biochemical electrolyte is realized.
(2) According to the microfluidic chip, the sample test groove is designed, the measurement volume is fixed, the measurement result is more accurate, meanwhile, the microfluidic chip and the electrode card can be separated by the design of the positioning column, the structure of the microfluidic chip is simplified, the production process difficulty and the production efficiency of the microfluidic chip are greatly reduced, the electrode card can be installed according to a detection target object, and the microfluidic chip is suitable for multiple index analysis in blood gas detection.
(3) The array electrode plate is loaded in the clamping groove of the microfluidic chip, a plurality of microelectrodes are integrated on the electrode plate, and the microfluidic sensing chip is combined, so that a plurality of biochemical molecules can be detected simultaneously, and the array electrode plate can be applied to multi-parameter electrochemical detection.
Drawings
The invention will be further described with reference to the accompanying drawings, in which embodiments do not constitute any limitation of the invention, and other drawings can be obtained by one of ordinary skill in the art without inventive effort from the following drawings.
Fig. 1 is a schematic structural diagram of a microfluidic detection card provided by the present invention;
fig. 2 is a schematic front view of a microfluidic detection card provided by the present invention;
fig. 3 is a schematic back view of a microfluidic detection card provided by the present invention;
FIG. 4 is a schematic diagram of the front structure of an electrode card;
FIG. 5 is a schematic view of the back structure of an electrode card;
fig. 6 is a schematic diagram of liquid sample injection of the microfluidic detection card provided by the invention.
The attached drawings are identified:
1. a detection card body; 11. an exhaust hole; 12. a detection card positioning hole;
2. a microfluidic system; 21. a sample inlet; 22. a pretreatment chamber; 23. a pre-treatment buffer channel; 24. a miniature check valve; 25. a diverter valve; 26. a calibration liquid bag; 27. an electrode assembly region; 28. a simple check valve; 29. a waste liquid tank;
3. an array electrode mounting clamping groove; 31. a sample test slot; 32. positioning columns;
4. an electrode card; 41. positioning holes;
51. a reference electrode; 52. an auxiliary electrode; 53. a working electrode A; 54. a working electrode B; 55. calibrating the electrodes.
Detailed Description
The invention will be further described in detail with reference to the following specific examples, with reference to the accompanying drawings, in order to make the objects, technical solutions and advantages of the invention more apparent.
Example 1
As shown in figure 1, the microfluidic detection card provided by the invention specifically comprises a detection card body 1, wherein the width of the detection card body is 70mm, the height of the detection card body is 70mm, and the thickness of the detection card body is 3.0mm.
In this embodiment, the sample inlet 21 on the main body 1 of the test card is provided with a sample inlet pipe inside, and the width is 1.0mm and the height is 1.0mm.
In this embodiment, the sample injection pipe system and the calibration fluid bag 26 are located inside the detection card body 1, and the width of the calibration fluid bag 26 is 13mm and the height is 31mm.
In this embodiment, the sample feeding pipe system includes a pretreatment chamber 22, a pretreatment buffer channel 23 and a micro check valve 24, which are sequentially connected to the sample feeding port, the electrode assembly area 27 is connected to the sample feeding pipe system through a sample feeding liquid channel buried in the detection card body 1, the sample feeding liquid channel is connected to the micro check valve 24, and a calibration liquid channel is extended from the micro check valve 24 to be connected to a calibration liquid bag 26, and a flow guiding valve 25 is disposed between the calibration liquid channel and the calibration liquid bag 26.
The pretreatment chamber 22 houses the immune microspheres.
The pretreatment buffer channel 23 is a micro flow channel formed by a plurality of micro saddle-shaped structures.
The width of the sample injection liquid channel and the calibration liquid channel is 1.0mm, and the height is 1.0mm.
In this embodiment, the electrode mounting area 27 and the liquid lead-out system of the back surface of the card body 1 are detected.
In this embodiment, one end of the electrode assembly area 27 is connected to the sample liquid channel, and the other end is connected to the liquid guiding-out system through the sample liquid channel buried in the detection card body 1; the liquid guiding system is connected with an exhaust hole 11.
In this embodiment, the electrode assembly area 27 is a region dug out on the test card body 1 for sample detection, and is an array electrode mounting card slot 3, which can be specifically divided into two rows of 6 card slots with identical specifications, each card slot 8 is provided with a groove with a fixed volume, as a sample test slot 31, and a positioning column 32 is disposed at a corner, where the positions of the sample test slot 31 and the positioning column 32 respectively correspond to the microelectrode detection sites and the positioning holes 41 on the electrode card 4.
As shown in fig. 4 to 5, five microelectrodes are integrated on the front surface of the electrode card 4, and include 1 reference electrode, 1 auxiliary electrode and 3 working electrodes, wherein 1 working electrode can be used as a calibration electrode. The device comprises a reference electrode 51, an auxiliary electrode 52, a working electrode A53, a working electrode B54 and a calibration electrode 55, wherein the surface of the reference electrode 51 is modified with silver-silver chloride, the surface of the auxiliary electrode 52 is modified with a platinum layer, the surface of the working electrode can be modified with a gold layer, and different sensitive substances can be modified according to detection indexes. Each electrode card 4 can detect two material indexes simultaneously.
The back of the electrode card 4 is provided with metal pins with the same number as the microelectrodes, and the microelectrodes are correspondingly connected with the metal pins through metal circuits or through hole metal circuits in the electrode card 4.
The surface of the detection card main body 1 is provided with 6 electrode cards 4, the front surfaces of the electrode cards 4 are downwards arranged in the array electrode mounting card slots 3, the microelectrodes are correspondingly embedded into the sample test slots 31, and the sample liquid channels sequentially pass through all the sample test slots 31 and are connected with the sample liquid channels, so that the liquid to be detected contacts with the microelectrodes when passing through the sample test slots 31; the back of the electrode card 4 is upward, the electrode pins are exposed, the electric signal change of the microelectrode can be collected through the connection electrode pins, and the concentration of the object to be detected, which is detected by the microelectrode, is detected by adopting an electrochemical method.
Each electrode plate 4 can detect 2 different target objects, and the whole microfluidic detection card can detect 12 different material indexes simultaneously, so that multi-parameter instant electrochemical detection is realized.
The sample test slot 31 is a fixed volume groove with a width of 8.1mm and a height of 1.5mm; the positioning column 32 is cylindrical and has a radius of 0.2mm; the individual card slot width of the array electrode mounting card slot 3 is 13.4mm and the height is 7.0mm.
In this embodiment, the liquid delivery system includes a simple check valve 28, a waste liquid tank 29, and a vent 11.
The simple check valve 28 is a tesla valve.
In this embodiment, the detection card body 1 is further provided with a detection card positioning hole 12 for fixing the detection card to a detection instrument or other devices, and the aperture is 1.0mm.
Example 2
The embodiment provides a microfluidic detection card with an array electrode card integrated by a plurality of microelectrodes for multi-parameter biochemical molecular detection, which specifically comprises a detection card main body 1, wherein the width of the detection card main body 1 is 70mm, the height of the detection card main body 1 is 70mm, and the thickness of the detection card main body is 3.0mm.
In this embodiment, the sample inlet 21 on the main body 1 of the test card is provided with a sample inlet pipe inside, and the width is 1.5mm and the height is 1.5mm.
In this embodiment, the sample feeding pipe system and the calibration fluid bag 26 are located on the front surface of the detection card body 1, and the width of the calibration fluid bag 26 is 12mm, and the height is 30mm.
In this embodiment, the sample feeding pipe system includes a pretreatment chamber 22, a pretreatment buffer channel 23 and a micro check valve 24, which are sequentially connected to the sample feeding port, the electrode assembly area 27 is connected to the sample feeding pipe system through a sample feeding liquid channel buried in the detection card body 1, the sample feeding liquid channel is connected to the micro check valve 24, and a calibration liquid channel is extended from the micro check valve 24 to be connected to a calibration liquid bag 26, and a flow guiding valve 25 is disposed between the calibration liquid channel and the calibration liquid bag 26.
The pretreatment chamber 22 houses an anticoagulant drug.
The pretreatment buffer channel 23 is a micro flow channel formed by a plurality of micro saddle-shaped structures.
The width of the sample injection liquid channel and the calibration liquid channel is 1.0mm, and the height is 1.0mm.
In this embodiment, the electrode mounting area 27 and the liquid lead-out system of the back surface of the card body 1 are detected.
In this embodiment, one end of the electrode assembly area 27 is connected to the sample liquid channel, and the other end is connected to the liquid guiding-out system through the sample liquid channel buried in the detection card body 1; the liquid guiding system is connected with an exhaust hole 11.
In this embodiment, the electrode assembly area 27 is a region dug out on the main body of the detection card for sample detection, and is an array electrode mounting card slot 3, which can be divided into two rows of 4 card slots with identical specifications, each card slot is provided with a groove with a fixed volume, as a sample test slot 31, and meanwhile, a positioning column 32 is disposed at a corner, and the positions of the sample test slot 31 and the positioning column 32 respectively correspond to the microelectrode detection sites and the positioning holes 41 on the electrode card 4.
Five microelectrodes are integrated on the front face of the electrode card 4, and comprise 1 reference electrode, 1 auxiliary electrode and 3 working electrodes, wherein the 1 working electrodes can be used as calibration electrodes. The device comprises a reference electrode 51, an auxiliary electrode 52, a working electrode A53, a working electrode B54 and a calibration electrode 55, wherein the surface of the reference electrode is modified with silver-silver chloride, the surface of the auxiliary electrode is modified with a platinum layer, the surface of the working electrode can be modified with a gold layer, and different sensitive substances can be modified according to detection indexes. Each electrode card 4 can detect two material indexes simultaneously.
The back of the electrode card 4 is provided with metal pins with the same number as the microelectrodes, and the microelectrodes are correspondingly connected with the metal pins through metal circuits or through hole metal circuits in the electrode card 4.
The surface of the detection card main body 1 is provided with 4 electrode cards 4, the front surfaces of the electrode cards 4 are downwards arranged in the array electrode mounting card slots 3, the microelectrodes are correspondingly embedded into the sample test slots 31, and the sample liquid channels sequentially pass through all the sample test slots 31 and are connected with the sample liquid channels, so that the liquid to be detected contacts with the microelectrodes when passing through the sample test slots 31; the back of the electrode card 4 is upward, the electrode pins are exposed, the electric signal change of the microelectrode can be collected through the connection electrode pins, and the concentration of the object to be detected, which is detected by the microelectrode, is detected by adopting an electrochemical method.
Each electrode plate can detect 2 different target objects, and the whole microfluidic detection card can detect 8 different material indexes simultaneously, so that multi-parameter instant electrochemical detection is realized.
The sample test slot 31 is a fixed volume groove with a width of 8.0mm and a height of 1.5mm; the positioning column is cylindrical, and the radius is 0.2mm; the individual card slot width of the array electrode mounting card slot 3 is 14.5mm, and the height is 7.5mm.
In this embodiment, the liquid delivery system includes a simple check valve 28, a waste liquid tank 29, and a vent 11.
The simple check valve 28 is a tesla valve.
In this embodiment, the detection card body 1 is further provided with a detection card positioning hole 12, and the aperture is 1.0mm.
Example 3
The embodiment provides a microfluidic detection card with an array electrode card integrated by a plurality of microelectrodes for multi-parameter biochemical molecular detection, which specifically comprises a detection card main body 1, wherein the width of the detection card main body 1 is 70mm, the height of the detection card main body 1 is 70mm, and the thickness of the detection card main body is 3.0mm.
In this embodiment, the sample inlet 21 on the main body 1 of the test card is provided with a sample inlet pipe inside, and the width is 1.5mm and the height is 1.5mm.
In this embodiment, the sample feeding pipe system and the calibration fluid bag 26 are located on the front surface of the detection card body 1, and the width of the calibration fluid bag 26 is 12mm, and the height is 30mm.
In this embodiment, the sample feeding pipe system includes a pretreatment chamber 22, a pretreatment buffer channel 23 and a micro check valve 24, which are sequentially connected to the sample feeding port, the electrode assembly area 27 is connected to the sample feeding pipe system through a sample feeding liquid channel buried in the detection card body 1, the sample feeding liquid channel is connected to the micro check valve 24, and a calibration liquid channel is extended from the micro check valve 24 to be connected to a calibration liquid bag 26, and a flow guiding valve 25 is disposed between the calibration liquid channel and the calibration liquid bag 26.
The pretreatment chamber 22 is empty.
The pretreatment buffer channel 23 is a micro flow channel formed by a plurality of micro saddle-shaped structures.
The width of the sample injection liquid channel and the calibration liquid channel is 1.5mm, and the height is 1.5mm.
In this embodiment, the electrode mounting area 27 and the liquid lead-out system of the back surface of the card body 1 are detected.
In this embodiment, one end of the electrode assembly area 27 is connected to the sample liquid channel, and the other end is connected to the liquid guiding-out system through the sample liquid channel buried in the detection card body 1; the liquid guiding system is connected with an exhaust hole 11.
In this embodiment, the electrode assembly area 27 is a region dug out on the main body of the detection card for sample detection, and is an array electrode mounting card slot 3, which can be specifically divided into a row of 3 card slots with identical specifications, each card slot is provided with a groove with a fixed volume, as a sample test slot 31, and meanwhile, a positioning column 32 is disposed at a corner, and the positions of the sample test slot 31 and the positioning column 32 respectively correspond to the microelectrode detection site and the positioning hole 41 on the electrode card 4, so that the electrode card 4 can be mounted and fixed in the sample test slot 31.
Five microelectrodes are integrated on the front face of the electrode card 4, and comprise 1 reference electrode, 1 auxiliary electrode and 3 working electrodes, wherein the 1 working electrodes can be used as calibration electrodes. The device comprises a reference electrode 51, an auxiliary electrode 52, a working electrode A53, a working electrode B54 and a calibration electrode 55, wherein the surface of the reference electrode is modified with silver-silver chloride, the surface of the auxiliary electrode is modified with a platinum layer, the surface of the working electrode can be modified with a gold layer, and different sensitive substances can be modified according to detection indexes. Each electrode card 4 can detect two material indexes simultaneously.
The back of the electrode card 4 is provided with metal pins with the same number as the microelectrodes, and the microelectrodes are correspondingly connected with the metal pins through metal circuits or through hole metal circuits in the electrode card 4.
The surface of the detection card main body 1 is provided with 3 electrode cards 4, the front surfaces of the electrode cards 4 are downwards arranged in the array electrode mounting card slots 3, the microelectrodes are correspondingly embedded into the sample test slots 31, and the sample liquid channels sequentially pass through all the sample test slots 31 and are connected with the sample liquid channels, so that the liquid to be detected contacts with the microelectrodes when passing through the sample test slots 31; the back of the electrode card 4 is upward, the electrode pins are exposed, the electric signal change of the microelectrode can be collected through the connection electrode pins, and the concentration of the object to be detected, which is detected by the microelectrode, is detected by adopting an electrochemical method.
Each electrode plate 4 can detect 2 different target objects, and the whole microfluidic detection card can detect 6 different material indexes simultaneously, so that multi-parameter instant electrochemical detection is realized.
The sample test slot 31 is a fixed volume groove with a width of 8.0mm and a height of 1.5mm; the positioning column is cylindrical, and the radius is 0.2mm; the width of the single electrode mounting clamping groove is 13.0mm, and the height is 6.5mm.
In this embodiment, the liquid delivery system includes a simple check valve 28, a waste liquid tank 29, and a vent 11.
The simple check valve 28 is a tesla valve.
In this embodiment, the detection card main body is further provided with a detection card positioning hole 12, and the aperture is 1.0mm.
Example 4
The embodiment provides a microfluidic detection card with an array electrode card integrated by a plurality of microelectrodes for multi-parameter biochemical molecular detection, which specifically comprises a detection card main body 1, wherein the width of the detection card main body 1 is 70mm, the height of the detection card main body 1 is 70mm, and the thickness of the detection card main body is 3.0mm.
In this embodiment, the sample inlet 21 on the main body 1 of the test card is provided with a sample inlet pipe inside, and the width is 1.5mm and the height is 1.5mm.
In this embodiment, the sample feeding pipe system and the calibration fluid bag 26 are located on the front surface of the detection card body 1, and the width of the calibration fluid bag 26 is 12mm, and the height is 30mm.
In this embodiment, the sample feeding pipe system includes a pretreatment chamber 22, a pretreatment buffer channel 23 and a micro check valve 24, which are sequentially connected to the sample feeding port, the electrode assembly area 27 is connected to the sample feeding pipe system through a sample feeding liquid channel buried in the detection card body 1, the sample feeding liquid channel is connected to the micro check valve 24, and a calibration liquid channel is extended from the micro check valve 24 to be connected to a calibration liquid bag 26, and a flow guiding valve 25 is disposed between the calibration liquid channel and the calibration liquid bag 26.
The pretreatment chamber 22 is empty.
The pretreatment buffer channel 23 is a micro flow channel formed by a plurality of micro saddle-shaped structures.
The width of the sample injection liquid channel and the calibration liquid channel is 1.5mm, and the height is 1.5mm.
In this embodiment, the electrode mounting area 27 and the liquid lead-out system of the back surface of the card body 1 are detected.
In this embodiment, one end of the electrode assembly area 27 is connected to the sample liquid channel, and the other end is connected to the liquid guiding-out system through the sample liquid channel buried in the detection card body 1; the liquid guiding system is connected with an exhaust hole 11.
In this embodiment, the electrode assembly area 27 is a region dug out on the test card body 1 for sample detection, in which an electrode mounting slot is provided, in which a fixed-volume groove is provided as the sample test slot 31, and positioning posts 32 are provided at corners, and the positions of the sample test slot 31 and the positioning posts 32 correspond to the microelectrode detection sites and the positioning holes 41 on the electrode card 4, respectively.
Five microelectrodes are integrated on the front face of the electrode card 4, and comprise 1 reference electrode, 1 auxiliary electrode and 3 working electrodes, wherein the 1 working electrodes can be used as calibration electrodes. The device comprises a reference electrode 51, an auxiliary electrode 52, a working electrode A53, a working electrode B54 and a calibration electrode 55, wherein the surface of the reference electrode is modified with silver-silver chloride, the surface of the auxiliary electrode is modified with a platinum layer, the surface of the working electrode can be modified with a gold layer, and different sensitive substances can be modified according to detection indexes. Each electrode card 4 can detect two material indexes simultaneously.
The back of the electrode card 4 is provided with metal pins with the same number as the microelectrodes, and the microelectrodes are correspondingly connected with the metal pins through metal circuits or through hole metal circuits in the electrode card 4.
The surface of the detection card main body 1 is provided with 1 electrode card 4, the front surface of the electrode card 4 is downwards arranged in the electrode mounting card slot, the microelectrode is correspondingly embedded into the sample test slot 31, and the sample liquid channel sequentially passes through all the sample test slots 31 and is connected with the sample liquid channel, so that the liquid to be detected contacts with the microelectrode when passing through the sample test slot 31; the back of the electrode card 4 is upward, the electrode pins are exposed, the electric signal change of the microelectrode can be collected through the connection electrode pins, and the concentration of the object to be detected, which is detected by the microelectrode, is detected by adopting an electrochemical method.
Each electrode plate 4 can detect 2 different target objects, and the whole microfluidic detection card can detect 2 different material indexes simultaneously, so that multi-parameter instant electrochemical detection is realized.
The sample test slot 31 is a fixed volume groove with a width of 8.0mm and a height of 1.5mm; the positioning column is cylindrical, and the radius is 0.2mm; the electrode mounting clamping groove has a width of 13.0mm and a height of 6.5mm.
In this embodiment, the liquid delivery system includes a simple check valve 28, a waste liquid tank 29, and a vent 11.
The simple check valve 28 is a tesla valve.
In this embodiment, the detection card body 1 is further provided with a detection card positioning hole 12, and the aperture is 1.0mm.
Example 5
According to the microfluidic chip for multi-parameter biochemical molecule detection provided in embodiment 1, as shown in fig. 6, a schematic liquid sample injection diagram of the microfluidic detection card provided in the invention is provided.
When in sample adding, the microfluidic detection card is horizontally placed, the front face is upward, liquid to be measured is injected from the sample inlet 21, flows through the pretreatment chamber 22, flows into the pretreatment buffer channel 23, flows through the micro check valve 24, simultaneously uses a mechanical device to squeeze the calibration liquid bag 26, squeezes the calibration liquid in the calibration liquid bag 26 to flow out, flows into the calibration liquid channel after passing through the flow guide valve 25, and is mixed with the liquid to be measured through the micro check valve 24; the mixed liquid sequentially passes through the sample liquid channel, the sample test groove 31 and the microelectrode which is contacted with the front surface of the electrode card 4, then passes through the sample liquid channel, flows through the simple check valve 28 and enters the waste liquid groove 29, gas generated in the process is discharged through the exhaust hole 11 connected with the waste liquid groove 29, and the detection electric signal is transmitted through the electrode pin of the electrode card 4.
The electrode mounting clamping groove is of an array design, 6 electrode cards with the same specification are mounted, five microelectrodes are modified on the front face of each electrode card, the positions of the microelectrodes correspond to the sample testing grooves 31, and the sample testing grooves 31 with continuous groove shapes are arranged in the array electrode mounting clamping groove 3 and are communicated with the sample injection liquid channel. The liquid to be tested sequentially flows through all sample test grooves 31 through the sample liquid channel and contacts with the microelectrodes corresponding to the positions of the sample test grooves, so that the aim of simultaneously detecting at most 12 different indexes is fulfilled.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present disclosure describes embodiments, not every embodiment is provided with a separate embodiment, and that this description is provided for clarity only, and that the disclosure is not limited to the embodiments described in detail below, and that the embodiments described in the examples may be combined appropriately to form other embodiments that will be understood by those skilled in the art. Technical details not described in detail in the present invention may be implemented by any prior art in the field. In particular, all technical features not described in detail in this invention can be realized by any prior art technique.
Claims (14)
1. The array electrode card with the integrated multiple microelectrodes is used for detecting multiparameter biochemical molecules and comprises a detection card main body (1), a microfluidic system (2) and an array electrode mounting clamping groove (3) which are arranged in the detection card main body (1), and an electrode card (4) which is loaded in the array electrode mounting clamping groove (3), wherein the surface of the electrode card (4) is integrated with multiple microelectrodes; the microfluidic system (2) comprises a sample inlet (21), a sample inlet pipeline system, a calibration liquid system, an electrode assembly area (27) and a liquid guiding-out system; the electrode assembly area (27) is provided with an array type electrode installation clamping groove (3), the array type electrode installation clamping groove (3) is formed by arranging a plurality of electrode installation clamping grooves in an array mode, a piece of electrode card (4) can be loaded in a single clamping groove, a plurality of microelectrodes are integrated on the front face of the electrode card (4), the back face of the electrode card (4) is provided with the same number of electrode pins, and the microelectrodes and the electrode pins are correspondingly connected with a metal circuit one by one through conducting sites on the surface of the electrode sheet (4); a sample test groove (31) is formed in each clamping groove of the array electrode mounting clamping groove (3), the front surface of the electrode card (4) is loaded in the clamping groove downwards, and microelectrodes modified on the surface of the electrode card (4) are correspondingly embedded in the sample test groove (31); one end of the electrode assembly area (27) is connected with the sample injection pipeline system, and the other end of the electrode assembly area is communicated with the liquid guiding-out system; the sample introduction pipe system passes through the electrode assembly area (27), specifically each clamping groove of the array electrode installation clamping groove (3), and is connected with the sample test groove (31).
2. The array electrode card with the integrated multiple microelectrodes for the multiparameter biochemical molecular detection according to claim 1, wherein the detection card main body (1) is provided with an exhaust hole (11), and the exhaust hole (11) is communicated with the liquid guiding-out system.
3. The microfluidic detection card for multi-parameter biochemical molecular detection using the array electrode card with the integrated multiple microelectrodes according to claim 1 is characterized in that the detection card main body (1) is further provided with a detection card positioning hole (12).
4. The array electrode card for multi-parameter biochemical molecular detection according to claim 1, wherein in the microfluidic system (2), the sample injection pipeline system comprises a pretreatment chamber (22), a pretreatment buffer channel (23), a micro check valve (24) and a sample injection liquid channel, the sample injection liquid channel is connected with the sample injection port (21) and is sequentially communicated with the pretreatment chamber (22), the pretreatment buffer channel (23) and the micro check valve (24), and the calibration liquid system is connected with the micro check valve (24).
5. The card for multi-parameter biochemical molecular detection according to claim 4, wherein the pretreatment chamber (22) is internally provided with immunomagnetic beads, anticoagulant drugs or empty.
6. The microfluidic detection card for multi-parameter biochemical molecular detection according to claim 4, wherein the pretreatment buffer channel (23) can be a micro-flow channel formed by a plurality of micro-saddle structures.
7. The array electrode card with the integrated multiple microelectrodes for the multiparameter biochemical molecular detection according to the claim 1, wherein in the microfluidic system (2), the calibration liquid system comprises a calibration liquid channel and a calibration liquid package (26), the calibration liquid package (26) is filled with the calibration liquid, and a flow guide valve (25) is arranged between the calibration liquid channel and the calibration liquid package (26); the calibration fluid channel is connected to the sample introduction tubing.
8. The array electrode card with the integrated multiple microelectrodes for the multi-parameter biochemical molecular detection according to claim 1 is characterized in that in the microfluidic system (2), the liquid guiding-out system comprises a simple check valve (28), a waste liquid tank (29) and a sample outlet liquid channel, wherein the sample outlet liquid channel is connected with the sample inlet pipeline system, and the sample outlet liquid channel is sequentially communicated with the simple check valve (28) and the waste liquid tank (29).
9. The microfluidic detection card for multiparameter biochemical molecular detection of claim 8, wherein the simple check valve (28) structure comprises a tesla valve structure.
10. The array electrode card for multi-parameter biochemical molecular detection of claim 1, wherein the sample test groove (31) is a groove with a fixed volume in the array electrode mounting groove (3).
11. The array electrode card with the integrated multiple microelectrodes for the multi-parameter biochemical molecular detection of claim 1 is characterized in that a positioning column (32) is arranged in each clamping groove of the array electrode mounting clamping groove (3), a positioning hole (41) is arranged on the surface of the electrode card (4), the positioning column (32) corresponds to the positioning hole (41) in position and is in a consistent size, and the electrode card (4) can be fixed inside the array electrode mounting clamping groove (8) through the positioning hole (41).
12. The array electrode card for multiparameter biochemical molecular detection according to claim 1, wherein the microelectrodes integrated on the surface of the electrode card (4) comprise a reference electrode, an auxiliary electrode and a working electrode, and the working electrode comprises a calibration electrode.
13. The microfluidic detection card for multiparameter biochemical molecular detection according to claim 12, wherein the microelectrodes of the electrode card (4) surface modification comprise a reference electrode (51), an auxiliary electrode (52), a working electrode a (53), a working electrode B (54) and a calibration electrode (55).
14. A method of using a multi-microelectrode integrated array electrode card for multiparameter biochemical molecular detection comprising the microfluidic detection card of any one of claims 1 to 13, comprising the steps of: when in sample adding, the detection card is horizontally placed, the front face is upward, liquid to be detected is injected from the sample inlet (21), flows through the pretreatment chamber (22), flows into the pretreatment buffer channel (23), flows through the miniature check valve (24), simultaneously uses a mechanical device or manpower to squeeze the calibration liquid bag (26), extrudes the calibration liquid in the calibration liquid bag (26) to flow out, flows into the calibration liquid channel after passing through the flow guide valve (25), and is mixed with the liquid to be detected in the sample injection liquid channel through the miniature check valve (24); the mixed liquid sequentially passes through a sample liquid channel, a sample test groove (31) and a microelectrode which is in contact with the front surface of the electrode card (4), then passes through a sample liquid channel, flows through a simple check valve (28) and enters a waste liquid groove (29), gas generated in the process is discharged through an exhaust hole (11) connected with the waste liquid groove (29), and a detection electric signal is transmitted through an electrode pin on the back surface of the electrode card (4).
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116539701A (en) * | 2023-07-07 | 2023-08-04 | 北京几何科技有限公司 | Reagent card for urine detection and detection method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116539701A (en) * | 2023-07-07 | 2023-08-04 | 北京几何科技有限公司 | Reagent card for urine detection and detection method |
| CN116539701B (en) * | 2023-07-07 | 2023-09-15 | 北京几何科技有限公司 | Reagent card for urine detection and detection method |
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