CN116351483A - Integrated microfluidic biological reaction/detection device - Google Patents
Integrated microfluidic biological reaction/detection device Download PDFInfo
- Publication number
- CN116351483A CN116351483A CN202210856892.XA CN202210856892A CN116351483A CN 116351483 A CN116351483 A CN 116351483A CN 202210856892 A CN202210856892 A CN 202210856892A CN 116351483 A CN116351483 A CN 116351483A
- Authority
- CN
- China
- Prior art keywords
- sample
- polystyrene tube
- biological reaction
- detection device
- micro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 71
- 238000006243 chemical reaction Methods 0.000 title claims abstract description 53
- 239000004793 Polystyrene Substances 0.000 claims abstract description 60
- 229920002223 polystyrene Polymers 0.000 claims abstract description 60
- 239000007788 liquid Substances 0.000 claims description 21
- 125000006850 spacer group Chemical group 0.000 claims description 14
- 238000002347 injection Methods 0.000 claims description 9
- 239000007924 injection Substances 0.000 claims description 9
- 239000000523 sample Substances 0.000 abstract description 70
- 239000012472 biological sample Substances 0.000 abstract description 3
- 238000005842 biochemical reaction Methods 0.000 abstract 1
- 230000036039 immunity Effects 0.000 abstract 1
- 238000010586 diagram Methods 0.000 description 17
- 238000002965 ELISA Methods 0.000 description 10
- 238000000034 method Methods 0.000 description 9
- 102000004169 proteins and genes Human genes 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- 230000002255 enzymatic effect Effects 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 7
- -1 polyethylene Polymers 0.000 description 7
- 239000004698 Polyethylene Substances 0.000 description 6
- 229920000573 polyethylene Polymers 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 229940088598 enzyme Drugs 0.000 description 4
- 238000003018 immunoassay Methods 0.000 description 4
- 239000007790 solid phase Substances 0.000 description 4
- 239000004365 Protease Substances 0.000 description 3
- 238000001917 fluorescence detection Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 238000002038 chemiluminescence detection Methods 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000002967 competitive immunoassay Methods 0.000 description 2
- 238000007599 discharging Methods 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 235000019419 proteases Nutrition 0.000 description 2
- 108090000526 Papain Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 150000001251 acridines Chemical class 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- DZBUGLKDJFMEHC-UHFFFAOYSA-N benzoquinolinylidene Natural products C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 238000005266 casting Methods 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000012531 culture fluid Substances 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001125 extrusion Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 239000013610 patient sample Substances 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000012723 sample buffer Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Images
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Dispersion Chemistry (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
Abstract
The invention provides an integrated microfluidic biological reaction/detection device, which comprises: the device comprises a polystyrene tube body, wherein a sample inlet and a micro-channel communicated with the sample inlet are arranged on the polystyrene tube body; the interval section is arranged at one end of the polystyrene tube body far away from the sample inlet; the sample adding device is arranged at one end of the interval section and is communicated with the micro-flow channel, so that a sample enters into or exits from the micro-flow channel. The invention takes the polystyrene tube as a main body, the sample inlet and the micro-channel are arranged in the polystyrene tube, and meanwhile, the interval section and the sample adding device are integrated on the polystyrene tube, so that the polystyrene tube can be used as a device for sample collection, biological/biochemical reaction and molecular immunity detection, has simple structure and convenient use, and has smaller required sample quantity, higher reaction efficiency and no pollution to biological samples.
Description
The present application claims priority from the chinese patent office, application No. 202111629391.X, chinese patent application entitled "integrated microfluidic biological reaction/detection device", filed 28, 12, 2021, the entire contents of which are incorporated herein by reference.
Technical Field
The invention relates to the technical field of biological detection, in particular to an integrated microfluidic biological reaction/detection device.
Background
Immunosensory technology (Immunoassay) is one of the most important categories in biosensing technology, and is also one of the most dependent detection technologies in life science research and medical detection at present. Specifically, immunosensory techniques refer to detection techniques that utilize the specific binding of antibodies to antigens to convert a specific molecular concentration in a liquid sample into a readable signal.
Currently, the more widely applied immunosensor technologies are all solid-phase immunodetection. Solid phase immunoassays, in turn, are most commonly used in competitive immunoassays (Competitive Immunoassay) and sandwich immunoassays (Sandwich Immunoassay, also known as double antibody sandwich methods). The sandwich method is mainly used for detecting larger biomolecules such as proteins, and is the most commonly used immunosensor technology in life science research and medical detection at present. The principle can be briefly described as that target proteins in a liquid sample (such as serum) are specifically adsorbed to the solid surface of a sensor through a Capture Antibody (Capture Antibody), and then a detection Antibody (Detection Antibody) with a certain label (such as an enzyme, a fluorescent probe, acridine esters or radioactive elements) is used to combine with target protein molecules specifically adsorbed to the surface of the sensor, so that the concentration of the target proteins in the sample can be indirectly determined through detecting the signal intensity brought by the biomarker molecules on the sensor.
The immunosensor technology most commonly used at present is the enzyme-linked immunosorbent assay (ELISA) developed in the 1980 to 1990 s, and the magnetic bead chemiluminescent assay (CLIA) popular after 2010. Wherein ELISA is mainly used for laboratory detection, and magnetic beads CLIA is mainly used for clinical diagnosis. ELISA is widely popularized in the field of life science research once being pushed out due to the characteristics of higher sensitivity, lower cost and safety without radiation risk, and has been popular in the field of medical detection once. Sample types that can be detected by ELISA include animal body fluids (serum, urine, cerebrospinal fluid, etc.), cell and tissue culture fluids, patient samples (e.g., serum), etc. Almost all aspects related to life sciences are covered.
Since ELISA technology was developed earlier, 96-well plates of larger size are still currently used as immunoreactors, with a relatively low surface area to volume ratio per well. Moreover, the conventional 96-well plate ELISA has the problems that the required sample size is larger, for example, at least 100 mu L, the detection time is longer, generally 4-6 hours, and the manual operations such as sample adding, sample changing and cleaning are more. These all greatly increase the difficulty of researchers obtaining critical biological information in trace samples. The CLIA technique solves the problem of a large number of manual operations using automated mechanical means, but also fails to solve the problem of a large required sample size (at least 100 μl).
Disclosure of Invention
In view of the above, the technical problem to be solved by the present invention is to provide an integrated microfluidic biological reaction/detection device, which requires a small amount of sample (10 μl or less), and is convenient to use.
The invention provides a microfluidic biological reaction/detection device, comprising:
the device comprises a polystyrene tube body, wherein a sample inlet and a micro-channel communicated with the sample inlet are arranged on the polystyrene tube body;
the interval section is arranged at one end of the polystyrene tube body far away from the sample inlet;
the sample adding device is arranged at one end of the interval section and is communicated with the micro-flow channel, so that a sample enters into or exits from the micro-flow channel.
Specifically, the invention provides an integrated microfluidic biological reaction/detection device, comprising:
the device comprises a polystyrene tube body, wherein a sample inlet and a micro-channel communicated with the sample inlet are arranged on the polystyrene tube body;
the interval section is arranged at one end of the polystyrene tube body far away from the sample inlet;
and the mechanical liquid pump is arranged at one end of the interval section and is communicated with the micro-flow channel.
In one embodiment, the fluidic channel is a straight-through fluidic channel.
In one embodiment, the through-flow microchannels have a circular cross-section and a diameter of 200-1000 μm, preferably 400-800 μm.
In one embodiment, the cross section of the through-flow microchannel is square or rectangular.
In one embodiment, the polystyrene tube has a circular cross section, and the diameter of the circular shape is 1-2 mm.
In one embodiment, the wall thickness of the polystyrene tube body is 0.2-0.6 mm.
In one embodiment, the polystyrene tube has a rectangular or square cross section.
In one embodiment, the polystyrene tube has a height of 15 to 30mm.
In one embodiment, the polystyrene tube comprises:
a conical sample injection buffer area provided with a sample injection port;
and the reaction/detection area is communicated with the sample introduction buffer area, and the micro-channel is arranged in the reaction/detection area.
In one embodiment, the mechanical liquid pump may be a piston liquid pump, which may include:
a cavity disposed on the spacer;
the piston is arranged in the cavity, and one end of the piston can enter the interval section;
the piston pull rod is arranged at one end of the piston;
the top end of the piston pull rod is provided with a driving device;
and the limiting device is arranged at one end of the cavity, which is far away from the spacing section.
In one embodiment, the driving means may be a magnet block.
In other embodiments, the mechanical liquid pump may be a negative pressure air bag type liquid pump, and may include:
a cavity disposed on the spacer;
and the negative pressure air bag is coated on the cavity.
In some embodiments, the negative pressure bladder functions to create negative pressure by squeezing to force the sample into or out of the microchannel. The shape of the negative pressure air bag is not particularly limited, and the negative pressure air bag can be a balloon or an air bag with other shapes.
The invention provides an integrated microfluidic biological reaction/detection device, which comprises: the device comprises a polystyrene tube body, wherein a sample inlet and a micro-channel communicated with the sample inlet are arranged on the polystyrene tube body; the interval section is arranged at one end of the polystyrene tube body far away from the sample inlet; the sample adding device is arranged at one end of the interval section and is communicated with the micro-flow channel, so that a sample enters into or exits from the micro-flow channel. The invention takes the polystyrene tube as a main body, the sample inlet and the micro-flow channel are arranged in the polystyrene tube, and meanwhile, the interval section and the sample adding device, such as a mechanical liquid pump, are integrated on the polystyrene tube to form an integrated structure, so that the integrated structure can be used as a device for sample collection, biological reaction and detection, and the integrated structure has the advantages of simple structure, convenient use, small required sample amount, high reaction efficiency and no pollution to biological samples.
Drawings
Fig. 1 is a schematic structural diagram of an integrated microfluidic biological reaction/detection device according to an embodiment of the present invention;
fig. 2 is a schematic diagram of an exploded structure of an integrated microfluidic biological reaction/detection device according to an embodiment of the present invention;
FIG. 3 is a schematic view of a longitudinal section of a polyethylene pipe according to an embodiment of the present invention;
FIG. 4 is a schematic cross-sectional view of a polyethylene pipe according to an embodiment of the present invention;
FIG. 5 is a schematic cross-sectional view of a polystyrene tube according to another embodiment of the present invention;
FIG. 6 is a schematic diagram of the structure of one well of a 96-well plate;
FIG. 7 is a schematic cross-sectional view of an apparatus according to an embodiment of the present invention;
FIG. 8 is a schematic diagram of a detection flow provided by the present invention;
FIG. 9 is a conceptual diagram of chemiluminescent detection using the device of the present invention;
FIG. 10 is a conceptual diagram of enzymatic fluorescence detection using the apparatus provided by the present invention;
FIG. 11 is a conceptual diagram of optical absorption detection using the apparatus provided by the present invention;
FIG. 12 is a schematic illustration of an enzymatic reaction process provided by the present invention;
fig. 13 is a schematic structural diagram of a microfluidic biological reaction/detection device according to an embodiment of the present invention.
Detailed Description
The invention provides an integrated microfluidic biological reaction/detection device, which comprises:
the device comprises a polystyrene tube body, wherein a sample inlet and a micro-channel communicated with the sample inlet are arranged on the polystyrene tube body;
the interval section is arranged at one end of the polystyrene tube body far away from the sample inlet;
the sample adding device is arranged at one end of the interval section and is communicated with the micro-flow channel, so that a sample enters into or exits from the micro-flow channel.
Referring to fig. 1 and 2, fig. 1 is a schematic structural diagram of an integrated microfluidic biological reaction/detection device provided by an embodiment of the present invention, and fig. 2 is a schematic structural diagram of an integrated microfluidic biological reaction/detection device provided by an embodiment of the present invention, wherein 11 is a polystyrene tube, 111 is a sample inlet, 113 is a sample buffer area, 114 is a reaction/detection area, 12 is a spacer, and 13 is a mechanical liquid pump.
The microfluidic biological reaction detection device provided by the invention takes the polystyrene tube body 11 as a main structure, and polystyrene plastic has stronger protein affinity and can be used for immunodetection without any additional chemical treatment.
Referring to fig. 3 and fig. 4, fig. 3 is a schematic longitudinal section structure of a polyethylene pipe according to an embodiment of the present invention, and fig. 4 is a schematic cross section structure of a polyethylene pipe according to an embodiment of the present invention.
The polyethylene pipe body 11 is divided into a sample injection buffer area 113 and a reaction/detection area 114, wherein a sample injection port 111 is arranged on the part of the sample injection buffer area 113, and the sample injection buffer area 113 is of a conical structure, so that sample injection is facilitated.
The reaction/detection zone 114 is provided with a micro flow channel 112 for performing a biological reaction on the sample. The micro flow channel 112 is directly connected with the micro flow channel, and carries out biological reaction on the sample and detects the biological reaction.
The cross section of the polyethylene pipe 11 is circular, and the cross section of the micro flow channel 112 is circular. In one embodiment, the diameter of the through-flow microchannels 112 is 200 to 1000 μm. In one embodiment, the diameter of the through micro flow channel 112 is 400-800 μm and the diameter of the polystyrene tube is 1-2 mm. In one embodiment, the diameter of the straight-through micro flow channel 112 is 600 μm and the diameter of the polystyrene tube is 1.4mm.
In one embodiment, the wall thickness of the polystyrene tube body is 0.2-0.6 mm. In one embodiment, the polystyrene tube has a wall thickness of 0.4mm.
In one embodiment, the polystyrene tube has a height of 15 to 30mm. In one embodiment, the polystyrene tube has a height of 22mm.
In other embodiments, the cross section of the through micro flow channel 112 may be square or rectangular, and the cross section of the polystyrene tube 11 may also be rectangular or square, as shown in fig. 5, fig. 5 is a schematic cross-sectional structure of the polystyrene tube according to other embodiments of the present invention, where fig. 5A is a cross section of the polystyrene tube is square, and a cross section of the through micro flow channel is circular; fig. 5B is a square cross section of the polystyrene tube and a square cross section of the through micro flow channel, and fig. 5C is a rectangle cross section of the polystyrene tube and a rectangle cross section of the through micro flow channel.
As compared with the 96-well plate, FIG. 6 is a schematic diagram showing the structure of one well of the 96-well plate, wherein the diameter of one well is 6mm, the height of one well is 10mm, after the sample is added, the sample can be captured only at the bottom of the well, and the surface area and the volume of the sample are smaller and are only 0.32mm -1 . In the device provided by the present invention, referring to fig. 7, fig. 7 is a schematic cross-sectional structure of the device provided by the embodiment of the present invention, which is a through microThe diameter of the flow channel is 0.6mm, the height is 22mm, after the sample is added, the sample is captured in the whole range of the micro flow channel, and the surface area volume ratio is large and can reach 5mm -1 Thus the average distance of the molecules moving to the surface of the solid-phase immunosensor is greatly shortened, and the speed of the solid-phase immune reaction process is greatly accelerated.
The microfluidic biological reaction detection device provided by the invention further comprises the interval section 12 arranged at one end of the polystyrene tube body 11 far away from the sample inlet, wherein the interval section 12 is used for separating the biological reaction/detection area from the mechanical liquid pump on one hand, and on the other hand, the automation of the detection device can be realized.
Specifically, the spacer 12 is a hollow cavity, so that the mechanical liquid pump 13 is communicated with the micro flow channel, and the sample enters and exits the micro flow channel under the pumping and discharging actions of the mechanical liquid pump.
In one embodiment, the spacer 12 has a cross-sectional area greater than the cross-sectional area of the polystyrene tube 11, i.e., the spacer 12 has a protruding outer surface with respect to the polystyrene tube 11, may be used to form a mechanical connection with a robotic arm and control system to enable automated inspection.
In one embodiment of the present invention, the polystyrene tube 11, the spacer 12 and the mechanical liquid pump 13 (other than the piston) may be cast in one piece from polystyrene material. In other embodiments, the polystyrene tube 11, the spacer 12, and the mechanical liquid pump 13 (other than the piston) may be machined separately and then spliced into a unitary structure.
The microfluidic biological reaction detection device provided by the invention further comprises a mechanical liquid pump 13 arranged at one end of the interval section 12, wherein the mechanical liquid pump 13 is communicated with the micro-flow channel 112 and is used for sucking or discharging a sample into or out of the micro-flow channel.
In one embodiment, the mechanical liquid pump 13 includes a cavity 131 disposed on the spacer 12, a piston head 132 capable of moving in the cavity 131 by drawing, a limiting device 133 disposed at one end of the cavity 131 for limiting the stroke of the piston 132, a piston rod 134 connected to the piston head 132, and a magnet 135 disposed at the top end of the piston rod 134.
The main function of the cavity 131 is to accommodate the piston head 132 and provide a movable space for the piston head 132, one end of which is communicated with the spacer 12, and the other end of which is provided with a limiting device 133. The magnet 135 is used as a driving device to drive the piston to move, so that automation of reaction or detection is realized. In other embodiments, the magnets may be replaced with other driving means.
In one embodiment, the cavity 131, the spacer 12 and the polystyrene tube 11 may be molded in one piece using one-shot casting of polystyrene.
The microfluidic biological reaction/detection device provided by the invention integrates sample adding, biological reaction and detection, can use smaller sample quantity for reaction and detection, is more convenient to use, and can not cause the problem of biological sample pollution.
The microfluidic biological reaction/detection device provided by the invention can be used for enzyme-linked immunosorbent assay, and the specific flow is similar to that of the traditional ELISA. As shown in fig. 8, fig. 8 is a schematic diagram of a detection flow provided by the present invention, taking a sandwich method for detecting protein molecules as an example, the specific flow is as follows:
1. sucking the capture antibody solution into the micro-channel, wherein polystyrene has protein affinity, so that the capture antibody can be coated on the inner surface of the micro-channel, and the non-binding sites are blocked by using protein;
2. sucking a sample containing target molecules into the micro-channel coated with the antibody to combine the target molecules with the capture antibody;
3. adding an enzyme-linked detection antibody solution (the antibody is coupled with an enzyme molecule in advance by a chemical method) to enable the antibody to fully react with an immune complex;
4. a suitable substrate is added to generate a readable signal in the microchannel. Corresponding cleaning procedures are needed between each step to remove molecules which are not stably bound on the surface of the micro-flow tube.
In addition, if a proper antibody is used, the second step and the third step can be combined into a whole, and the sample and the ELISA antibody are premixed and then are simultaneously sucked into the microfluidic reactor.
The microfluidic biological reaction/detection device provided by the invention can be suitable for chemiluminescence detection, as shown in fig. 9, and fig. 9 is a conceptual schematic diagram of chemiluminescence detection by using the device provided by the invention; also suitable for enzymatic fluorescence detection, as shown in FIG. 10, FIG. 10 is a conceptual diagram of enzymatic fluorescence detection using the apparatus provided by the present invention; the device is also suitable for enzymatic chromogenic detection, as shown in FIG. 11, and FIG. 11 is a conceptual diagram of optical absorption detection by using the device provided by the invention. The detection method is not particularly limited, and the detection method is selected according to the selected detection method and the added detection substance. For example, chemiluminescent detection includes, but is not limited to, luminol chemiluminescence, acridine ester chemiluminescence; enzymatic chromogenic detection can measure absorbance at a specific wavelength, and the like.
In addition, the microfluidic biological reaction/detection device provided by the invention can be also used for a high-efficiency enzymatic synthesis/enzymatic preparation process of a trace sample, and the catalytic enzyme is fixed on the inner surface of the reactor. Taking an enzymatic proteolysis process as an example, referring to fig. 12, fig. 12 is a schematic diagram of an enzymatic reaction process provided in the present invention, and the specific flow is as follows:
1. coating protease (such as papain, trypsin, etc.) on the inner surface of the micro-channel;
2. the reaction raw material, i.e., the aqueous solution containing the protein, is inhaled. Waiting for a period of time to enable the protease fixed on the surface of the micro-channel to fully react with the protein solution;
3. the reaction product, i.e., the polypeptide solution, was drained and collected.
The device can be operated in a multi-tube parallel connection (such as a transverse arrangement or a matrix arrangement) mode when in actual use. In addition, the device can be directly combined with other biological culture systems (such as microfluidic organ chips and the like) to be used as a sample collection module.
In addition to the liquid pump with the piston and pull rod structure, the sample adding device may be a negative pressure air bag, for example, a balloon, see fig. 13, and fig. 13 is a schematic structural diagram of a microfluidic biological reaction/detection device according to an embodiment of the present invention. The difference compared to the microfluidic bio-reaction/detection device described above is that the balloon 23 is used instead of a piston and a pull rod for sample addition or sample ejection. In one embodiment, the balloon 23 is sleeved on the spacer, and negative pressure is generated by extrusion to enable the sample to enter the micro-channel. In a specific implementation manner, the polystyrene tube body, the interval section and the balloon can be processed respectively and then spliced into an integral structure, and the invention is not repeated.
The foregoing is only a preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art, who is within the scope of the present invention, should make equivalent substitutions or modifications according to the technical scheme of the present invention and the inventive concept thereof, and should be covered by the scope of the present invention.
Claims (10)
1. A microfluidic biological reaction/detection device comprising:
the device comprises a polystyrene tube body, wherein a sample inlet and a micro-channel communicated with the sample inlet are arranged on the polystyrene tube body;
the interval section is arranged at one end of the polystyrene tube body far away from the sample inlet;
the sample adding device is arranged at one end of the interval section and is communicated with the micro-flow channel, so that a sample enters into or exits from the micro-flow channel.
2. The integrated microfluidic biological reaction/detection device of claim 1, wherein the sample addition device is a mechanical liquid pump.
3. The microfluidic biological reaction/detection device of claim 2 wherein the fluidic channel is a straight-through fluidic channel.
4. A microfluidic biological reaction/detection device according to claim 3, wherein the cross section of the through-flow microchannel is circular, the diameter of the through-flow microchannel being 200-1000 μm, preferably 400-800 μm.
5. A microfluidic biological reaction/detection device according to claim 3, wherein the cross section of the through-flow microchannel is square or rectangular.
6. The microfluidic biological reaction/detection device according to any one of claims 1 to 5, wherein the polystyrene tube has a circular cross section, and the diameter of the circular shape is 1 to 2mm.
7. The microfluidic biological reaction/detection device according to claim 6, wherein the polystyrene tube has a tube wall thickness of 0.2-0.6 mm.
8. The microfluidic biological reaction/detection device according to any one of claims 1 to 5, wherein the polystyrene tube has a rectangular or square cross section;
the height of the polystyrene tube body is 15-30 mm.
9. The microfluidic biological reaction/detection device according to any one of claims 1 to 5, wherein the polystyrene tube comprises:
a conical sample injection buffer area provided with a sample injection port;
and the reaction/detection area is communicated with the sample introduction buffer area, and the micro-channel is arranged in the reaction/detection area.
10. The microfluidic biological reaction/detection device of claim 2, wherein the mechanical liquid pump comprises:
a cavity disposed on the spacer;
the piston is arranged in the cavity, and one end of the piston can enter the interval section;
the piston pull rod is arranged at one end of the piston;
the top end of the piston pull rod is provided with a driving device;
the limiting device is arranged at one end of the cavity, which is far away from the spacing section;
or,
the mechanical liquid pump comprises:
a cavity disposed on the spacer;
and the negative pressure air bag is coated on the cavity.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111629391 | 2021-12-28 | ||
| CN202111629391X | 2021-12-28 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116351483A true CN116351483A (en) | 2023-06-30 |
Family
ID=86926087
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210856892.XA Pending CN116351483A (en) | 2021-12-28 | 2022-07-20 | Integrated microfluidic biological reaction/detection device |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116351483A (en) |
-
2022
- 2022-07-20 CN CN202210856892.XA patent/CN116351483A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108535239B (en) | Micro-fluidic chip and detection system based on micro-droplets | |
| EP1776585B1 (en) | Immunoassay assembly and methods of use | |
| EP3779439B1 (en) | Micro-fluidic chip and analysis instrument having same | |
| Sasso et al. | Automated microfluidic processing platform for multiplexed magnetic bead immunoassays | |
| US9086409B2 (en) | Immunoassay biochip | |
| WO2019205778A1 (en) | Chemiluminescence microfluidic chip and analytical instrument having same | |
| US20060257956A1 (en) | Method and device for chemical or biological analysis by a sensor with a monolithic chamber in the form of a multi-microtubular sheaf and a lateral integration measuring transducer | |
| CN108761055B (en) | Microfluidic chip and analytical instrument with same | |
| US11904313B2 (en) | Liquid quantifying device and application thereof | |
| CN207703876U (en) | It is a kind of based on micro-fluidic fluorescence immunoassay detection chip | |
| US20220184619A1 (en) | Magnetic particle luminescent micro-fluidic chip for multi-marker detection and detection apparatus | |
| US20220357320A1 (en) | Two-layer microfluidic chip with magnetic bead luminescence and detection system | |
| CN110646609A (en) | Multi-marker detection magnetic particle luminous micro-fluidic chip and detection device | |
| CN113607704A (en) | Micro-fluidic chip detection method based on magnetic bead uniform mixing | |
| Torul et al. | Microfluidic-based blood immunoassays | |
| CN211374779U (en) | Multi-marker detection magnetic particle luminous micro-fluidic chip and detection device | |
| US7858047B2 (en) | Fluidic device | |
| CN116351483A (en) | Integrated microfluidic biological reaction/detection device | |
| CN211603208U (en) | Multi-marker detection magnetic particle luminous micro-fluidic chip and detection device | |
| CN104062445A (en) | Microfluid chip detection technique based on photonic crystal encoded microsphere | |
| RU200301U1 (en) | MICROFLUID CHIP FOR MULTI-PARAMETRIC IMMUNO ASSAY | |
| US20220241788A1 (en) | Analytical Device And Reaction Chamber | |
| Mehta et al. | Laboratory‐on‐a‐Chip: A Multitasking Device | |
| CN115840041A (en) | Quick detection device of disease marker | |
| CN117778174A (en) | Microfluidic chip for simultaneously performing molecular diagnosis, immunodiagnosis and biochemical detection and testing method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |