CN114405568B - Self-driven micro-fluidic chip - Google Patents
Self-driven micro-fluidic chip Download PDFInfo
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- CN114405568B CN114405568B CN202210206192.6A CN202210206192A CN114405568B CN 114405568 B CN114405568 B CN 114405568B CN 202210206192 A CN202210206192 A CN 202210206192A CN 114405568 B CN114405568 B CN 114405568B
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- 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
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- 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
Abstract
The invention provides a self-driven microfluidic chip for immunofluorescence detection, which mainly comprises a sample injection region for injecting target analytes; an antibody release region for releasing the fluorescent substance-antibody complex to bind to the target analyte; the channel detection area is used for capturing fluorescent substance-antibody-antigen complex formed after the sample passes through the antibody release area so as to carry out fluorescence detection; the device is characterized in that a structure which enables the channel to be bent up and down integrally exists at the front section of the channel detection area part, namely the channel is up and down fluctuated from the outside, so that liquid in the channel can generate horizontal displacement and up and down displacement when passing through the bent structure, a series of vortex flow fields are generated, reactants are fully and uniformly mixed, and the time for fluid to pass through can be adjusted by setting the number and the length of the bent structure.
Description
The invention relates to a micro-fluidic chip, in particular to a self-driven micro-fluidic chip for immunofluorescence detection.
Background
With the development of immune technology, point of care (POCT) is receiving attention and importance as a new field in test medicine. The method is a new method for immediately sampling and analyzing on the site of a patient, omits a complex processing procedure of a specimen in laboratory test, and rapidly obtains a test result, and has great significance in emergency rescue and the like. Has wide application in blood sugar detection, cardiovascular disease detection, infectious disease, drug abuse, pregnancy monitoring and other aspects. The POCT technology principle mainly comprises: colloidal gold, dry immunofluorescence, microfluidic immunofluorescence, and the like. At present, colloidal gold and dry immunofluorescence become the main stream technology in the market, and microfluidics is the main stream direction of future development.
Colloidal gold and dry immunofluorescence detection techniques are immunochromatography-based techniques, and generally require nitrocellulose membranes as a substrate for lateral flow of liquid. Their common advantages are simple and quick operation, high sensitivity and specificity. The biggest challenges faced by such immunochromatography techniques are poor reproducibility, primarily due to the need to adhere together a variety of thin, fragile materials while ensuring consistency in the flow of fluid therein. The solution idea is to thoroughly discard the chromatographic membrane and other films and adopt a micro-fluidic chip.
The micro-fluidic chip is based on micro-processing, and uses a micro-channel network as a structural feature, and does not need to resort to chromatography media, and the micro-fluidic chip aims to integrate part or all of operation units such as sampling, pretreatment, reagent adding, mixing, reaction, separation, detection and the like in the biological, chemical and medical analysis process on a chip with a micro-scale channel. The volume of the device is light, and the liquid directly flows in the air; the amount of the sample and the reagent is small; the reaction speed is high, and a large amount of parallel treatment can be realized. The self-driven microfluidic chip adopts capillary force as a driving force source, does not need equipment such as an auxiliary external air pump and the like, is simple and reliable to use, and becomes the main stream direction of immunofluorescence POCT detection development.
The main working steps of the microfluidic chip for immunofluorescence detection of antigen detection based on the double-sandwich principle are that 1, a sample is injected into a sample injection part; 2. the antigen in the sample is combined with the fluorescent-antibody complex of the antibody release region to form a fluorescent substance-antibody-antigen complex; 3. in the channel part, the fluorescent substance-antibody-antigen complex formed in the antibody releasing area is captured by the antibody pre-spotted in the detection area of the channel part, so that the substance with double-antibody sandwich of the fluorescent substance-antibody-antigen-antibody is formed in the detection area, and the amount of the antigen can be calculated by detecting the fluorescence intensity of the area by the fluorescent detection device. Each working step in the microfluidic chip has the problem of combination efficiency, and the combination efficiency of each step directly influences the detection result. The combination efficiency is related to factors such as the mixing degree of the sample, the reaction time and the like, and meanwhile, the quantity of bubbles and the flatness of the liquid level flow can influence the final detection efficiency.
At present, a self-driven microfluidic chip technology for clinical immunofluorescence detection is mainly a microfluidic chip technology owned by korea nanno technology limited, and patent (issued publication number: CN 102305867B) discloses a "chip for analyzing a fluid that does not use external energy for movement", which is designed as a self-driven microfluidic chip for analysis, and is divided into three parts, i.e., a preprocessing part, a channel part and a cleaning part, wherein the preprocessing part realizes the contents of steps 1 and 2 in the main steps of the operation of the microfluidic chip, and the channel part realizes the contents of step 3. The pretreatment device is mainly characterized in that a sample injection part and a first buffer part are arranged in the pretreatment part, and a step difference exists between the first phase buffer part and the sample injection part; the pretreatment part is also provided with a sample guide part which is arranged between the sample injection part and the first buffer part and used for destroying the surface tension of the fluid from the sample injection part to the first buffer part so as to achieve the purpose of stabilizing the fluid. The main invention point of the invention is concentrated between the injection of the sample into the first buffer part, and the relative design ensures that the fluid in the part can be more stable. However, the invention lacks an effective structural design for the channel part from the first buffer part to the second buffer part, so that the sample is fully mixed, various substances are fully combined, and fewer bubbles and the flatness of the liquid surface flow are ensured.
Therefore, it is necessary to design a novel self-driven microfluidic chip for immunofluorescence detection, and each part for realizing the functions is designed accurately, so that the related combination efficiency and the mixing degree are improved, the reaction time is regulated and controlled, the air bubbles are reduced, and the flatness of the liquid surface flow is ensured.
Disclosure of Invention
A self-driven microfluidic chip for immunofluorescence detection, comprising: a sample injection zone for injecting a target analyte; an antibody release region for releasing the fluorescent substance-antibody complex to bind to the target analyte; the channel detection area is used for capturing fluorescent substance-antibody-antigen complex formed after the sample passes through the antibody release area so as to carry out fluorescence detection; in the front section of the channel detection area part, a structure is arranged to bend the channel up and down, so that when the liquid in the channel passes through the bending structure, not only horizontal displacement is generated, but also up and down displacement is generated, thereby generating a series of vortex flow fields, and fully and uniformly mixing reactants.
The self-driven micro-fluidic chip for immunofluorescence detection is characterized in that the height of the front end of an antibody release area is gradually increased, fluid can slow down the flow velocity in the height range of the channel, the larger channel height can effectively inhibit the excessive bending of a flow surface, and the generation of bubbles in the fluid is reduced.
The self-driven microfluidic chip for immunofluorescence detection is characterized in that an exhaust hole is formed near the central axis of the tail end of an antibody release area and is used for reducing bubbles generated by the height difference and the surface tension effect of liquid in the flowing process of the antibody release area.
The self-driven microfluidic chip for immunofluorescence detection is characterized in that the initial height range of the front end of the antibody release region is 50-100 μm, and the termination height range is 100-500 μm.
The self-driven microfluidic chip for immunofluorescence detection is characterized in that the number of the bending structures at the front section of the channel detection area can be 1 or more.
The self-driven microfluidic chip for immunofluorescence detection is characterized in that the vertical displacement generated by the bending structure at the front section of the channel detection area can be adjusted as required, and the range of the value can be 100-500 mu m.
The beneficial effects of the invention are as follows:
1. under the condition of no external drive, the structure that the channel is vertically bent is designed in the channel detection area, so that fluid flows in the channel in a vertically bent mode, a vortex flow field is induced in the channel detection area, the efficiency of antigen-antibody binding reaction is enhanced, bubbles are effectively avoided, the diversion effect of the channel is ensured, in addition, the structure increases the volume of the channel, and the time required for binding before the antigen-antibody enters the detection area is fully ensured.
2. The fluid injected into the antibody release area is slowly changed along with the elevation of the fluid, so that sufficient time is provided for the release of the antibody, the fluid is fully evolved into a vortex structure flow field which is favorable for the diffusion and uniform mixing of the antibody in the antibody release area, and the surface tension provided by the upper wall, the lower wall and the side wall of the flow channel plays a key role in the formation of the vortex flow field.
3. Surface tension can cause the liquid flow velocity near the sidewall to be faster than in the middle region, resulting in the formation of a non-planar flow surface. The larger channel height of the antibody release area can effectively inhibit excessive bending of the flow surface and reduce the generation of bubbles in the fluid, thereby simplifying the design of the exhaust structure.
4. The exhaust holes near the central axis of the tail end of the antibody release area can effectively reduce the possibility of generating bubbles due to the height difference and the surface tension effect in the process of flowing liquid from the antibody release area into the channel detection area.
Drawings
Fig. 1 is a schematic view of the overall structure of a flow channel of a microfluidic chip according to the present invention.
Fig. 2 is a liquid velocity flow diagram in a software simulated microfluidic chip of the invention.
Fig. 3 is a liquid velocity flow diagram in a bump structure in a microfluidic chip of the present invention simulated by software.
Fig. 4 is a schematic diagram of the promotion of the raised structures in the microfluidic chip of the present invention to the flatness of the liquid surface by software simulation.
Detailed Description
The following describes the specific implementation method with reference to the accompanying drawings:
fig. 1 is a schematic view of the overall structure of a microfluidic chip flow channel according to the present invention, which is divided into 4 parts, namely, a 110 sample injection region, a 120 antibody release region, a 130 channel detection region, and a 140 waste liquid region front end. The whole display part is a runner of the micro-fluidic chip, namely, the sample enters the micro-fluidic chip from 110 and sequentially passes through the front ends of the 120 antibody release area, the 130 channel detection area and the 140 waste liquid area. In the figure: 110 is a sample injection zone for injecting a target analyte, on which a plurality of microcolumns are distributed for enlarging the specific surface area, enhancing the mixing of the sample and the preset at 110. The part 120 is an antibody release region, wherein 121 is the front end of the antibody release region, which plays a role of diversion, the front end of the antibody release region has a starting height ranging from 50 μm to 100 μm and a stopping height ranging from 100 μm to 500 μm, and the height of the liquid in 121 is gradually increased, and the flow rate is slowed down, thereby providing sufficient time for antibody release. 122 is a sample application area of the fluorescent substance-antibody complex, in this part of the area, the antigen to be tested can be fully mixed and combined with the fluorescent substance-antibody complex to form the fluorescent substance-antibody-antigen complex, in this part, the mixing degree and mixing time of the two substances are critical factors, and the related requirements can be met through the structural design. And 123 is an exhaust hole near the central axis of the tail end of the antibody release area, and is used for reducing bubbles generated by the height difference and the surface tension effect of the liquid in the flowing process of the antibody release area. 130 is a channel detection zone for capturing fluorescent material-antibody-antigen complexes formed after the sample passes through the antibody release zone, thereby performing fluorescence detection. The structure 131 is a structure in the front section of the channel detection area, which makes the channel bend up and down, i.e. the channel appears up and down from the outside, so that when the liquid in the channel passes through the bending structure, not only horizontal displacement is generated, but also up and down displacement is generated, the structure can be designed into one or more up and down structure according to the relevant properties of reactants, such as viscosity, etc., the flowing time of the fluid in the channel is directly related to the number of the structure and the up and down height of the structure, the up and down displacement can be adjusted as required, and the value range can be 100 μm to 500 μm, thereby playing the role of adjusting the passing time of the fluid. 140 is the front end of the waste liquid zone, and the shape and size of the waste liquid zone behind it can be freely set depending on how much liquid is desired to be contained.
Fig. 2 is a liquid velocity flow diagram in a microfluidic chip of the present invention simulated in software using a finite element based method. The lines in the figure represent velocity streamlines, i.e. the flow trend of the fluid in the flow channel. The fluid injected into the antibody release region 120, as it rises in height at 121, slows down, provides sufficient time for antibody release, and evolves sufficiently at 122 into a vertically symmetric vortex structured flow field that facilitates diffusion of the fluorescent material-antibody complex and mixing with the analyte. The surface tension provided by the upper and lower walls and the side walls of the flow channel plays a key role in the formation of the vortex flow field. At the up-and-down fluctuation convex structure position of 131, two similar vortex flow fields are generated by taking the symmetry axis of the micro-fluidic chip as the symmetry center, so that the mixing and combination of the fluorescent substance-antibody complex and the object to be detected are further enhanced.
Fig. 3 is a flow chart of liquid velocity in an up-and-down curved structure 131 of a microfluidic chip of the present invention, which has 2 convex structures curved up-and-down, using software simulation based on a finite element method, as a side view. It can be seen from the figure that, in the interior of the convex structure, particularly in the rising part thereof, a stronger vortex flow field is generated, and the mixing and combination of the fluorescent substance-antibody complex and the object to be detected are enhanced, so that the combination efficiency is improved, the detection sensitivity is improved, and in practical application, the convex structure can be designed according to practical requirements.
Fig. 4 is a schematic diagram of the speed promotion effect of a bump structure in a microfluidic chip of the present invention on the flatness of a liquid surface in a software simulation. The flow lines in the figure are velocity flow lines in the liquid, with the front most level of the fluid indicated by arrows. FIG. 4 (a) is a channel that does not include a raised structure; fig. 4 (b) is a channel comprising two raised structures. As can be seen from the figure, for the microfluidic chip with the convex structure according to the present invention, the flow velocity of the liquid is reduced as the height of the portion 131 increases, so that the liquid level (the portion indicated by the arrow in the figure) at the convex portion tends to be perpendicular to the flow channel direction, i.e. a flat liquid level is formed, which can effectively avoid bubble generation, and thus no additional exhaust structure is required to be designed in the channel detection region 130. As can be seen from the previous simulation of the microfluidic chip without the bump structure, the liquid surface (the part indicated by the arrow in the figure) is curved, and is relatively flat, so that bubbles are more likely to be generated.
Claims (5)
1. A self-driven microfluidic chip for immunofluorescence detection, comprising:
a sample injection zone for injecting a target analyte;
an antibody release region for releasing the fluorescent substance-antibody complex to bind to the target analyte;
the height of the front end of the antibody release area is gradually increased, the flow speed of the fluid can be slowed down within the height range of the channel, the excessive bending of the flow surface can be effectively restrained, and the generation of bubbles in the fluid can be reduced;
the channel detection area is used for capturing fluorescent substance-antibody-antigen complex formed after the sample passes through the antibody release area so as to carry out fluorescence detection;
in the front section of the channel detection area part, a structure is arranged to bend the channel up and down, so that when the liquid in the channel passes through the bending structure, not only horizontal displacement is generated, but also up and down displacement is generated, thereby generating a series of vortex flow fields, and fully and uniformly mixing reactants.
2. The self-driven microfluidic chip for immunofluorescence detection according to claim 1, wherein an exhaust hole is provided near the central axis of the end of the antibody release region for reducing bubbles generated by the height difference and the surface tension effect of the liquid during the flowing of the antibody release region.
3. The self-driven microfluidic chip for immunofluorescence detection according to claim 1, wherein the front end of the antibody release region has a starting height ranging from 50 μm to 100 μm and a terminating height ranging from 100 μm to 500 μm.
4. The self-driven microfluidic chip for immunofluorescence detection according to claim 1, wherein the number of bent structures at the front section of the channel detection region is 1 or more.
5. The self-driven microfluidic chip for immunofluorescence detection according to claim 1, wherein the vertical displacement generated by the curved structure at the front section of the channel detection region can be adjusted as required, and the range of the displacement can be 100 μm to 500 μm.
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| EP3229963A1 (en) * | 2014-12-12 | 2017-10-18 | Opko Diagnostics, LLC | Fluidic systems comprising an incubation channel, including fluidic systems formed by molding |
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| CN111013677A (en) * | 2019-12-23 | 2020-04-17 | 东莞市东阳光诊断产品有限公司 | Microfluidic chip, detection device and detection method |
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| CN108080042A (en) * | 2017-11-13 | 2018-05-29 | 成都微康生物科技有限公司 | Micro-fluidic chip of binding time resolved fluorometric technology and its preparation method and application |
| CN107974392A (en) * | 2017-12-27 | 2018-05-01 | 深圳市合川医疗科技有限公司 | A kind of method of circulating tumor cell in micro-fluidic chip and separating trap blood |
| US20210094033A1 (en) * | 2018-04-26 | 2021-04-01 | Nordetect Ivs | A microfluidic detection device with immobilized biochemical assays, fabrication of same and method of analysing a fluid sample |
| CN208894245U (en) * | 2018-06-26 | 2019-05-24 | 宁波奥丞生物科技有限公司 | The micro-fluidic chip of rapid quantitative detection PLGF and sFLT-1 |
| CN109499634B (en) * | 2018-12-13 | 2023-11-03 | 迪亚莱博（张家港）生物科技有限公司 | Microfluidic chip and preparation method and detection method thereof |
| CN215066717U (en) * | 2021-02-23 | 2021-12-07 | 宁波奥丞生物科技有限公司 | NT-proBNP quantitative detection micro-fluidic chip |
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Patent Citations (3)
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| EP3229963A1 (en) * | 2014-12-12 | 2017-10-18 | Opko Diagnostics, LLC | Fluidic systems comprising an incubation channel, including fluidic systems formed by molding |
| CN108686727A (en) * | 2018-06-26 | 2018-10-23 | 宁波奥丞生物科技有限公司 | The micro-fluidic chip of Quantitative detection PLGF and sFLT-1 |
| CN111013677A (en) * | 2019-12-23 | 2020-04-17 | 东莞市东阳光诊断产品有限公司 | Microfluidic chip, detection device and detection method |
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