CN111889154A - High-flux multi-target microfluidic biochip based on three-dimensional plasmon metamaterial - Google Patents
High-flux multi-target microfluidic biochip based on three-dimensional plasmon metamaterial Download PDFInfo
- Publication number
- CN111889154A CN111889154A CN202010784687.8A CN202010784687A CN111889154A CN 111889154 A CN111889154 A CN 111889154A CN 202010784687 A CN202010784687 A CN 202010784687A CN 111889154 A CN111889154 A CN 111889154A
- Authority
- CN
- China
- Prior art keywords
- biochip
- sensing device
- dimensional
- dimensional plasmon
- microfluidic
- 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
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
-
- 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
- B01L3/502707—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 characterised by the manufacture of the container or its components
-
- 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
- B01L3/50273—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 characterised by the means or forces applied to move the fluids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
- B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/06—Auxiliary integrated devices, integrated components
- B01L2300/0627—Sensor or part of a sensor is integrated
Abstract
The invention discloses a high-flux multi-target microfluidic biochip based on a three-dimensional plasmon metamaterial, which comprises: a biochip; the biochip includes: the device comprises a plastic cover plate, a three-dimensional plasmon metamaterial sensing device, a microfluidic bottom plate and a plurality of biological reagents; the plastic cover plate, the three-dimensional plasmon metamaterial sensing device and the microfluidic bottom plate are compounded from top to bottom, wherein the plastic cover plate is adhered to the microfluidic bottom plate by an adhesive, and the three-dimensional plasmon metamaterial sensing device is embedded in a pit of a detection area of the microfluidic bottom plate; the biological reagents include, but are not limited to, 11-mercaptoundecanoic acid solution MUA, ethyldimethylamine propyl carbodiimide EDC, N-hydroxysuccinimide NHS, bovine serum albumin BSA, ligand solution, and analyte solution. The high-flux multi-target microfluidic biochip based on the three-dimensional plasmon metamaterial can provide 24-channel high-flux multi-marker parallel real-time detection.
Description
Technical Field
The invention relates to the technical field of biochips and plasmon metamaterials, in particular to a high-flux multi-target microfluidic biochip based on a three-dimensional plasmon metamaterials.
Background
The application of the biomolecule sensing based on the spectrum detection technology has very important practical significance for various fields such as medical inspection, biological analysis, drug research and development, chemistry and chemical engineering, food safety, environmental monitoring and the like. In the last decade, a biomolecule sensing technology using surface plasmon optical effect shows extremely high sensitivity and resolution, and has the advantages of real-time monitoring, non-invasive detection, label-free and the like, so that the biomolecule sensing technology is generally concerned by scientists and technical developers in related fields. For example, the Biacore series of products and biochips under the american general medical flag have become an industry standard in the field of protein molecule interactions and kinetic analysis. Although commercial surface plasmon biosensing analysis techniques based on prism coupling schemes are mature. However, the biochip has many disadvantages of high price, complicated optical path, complicated operation, limited size of detection molecules, huge volume of detection equipment and the like, and is not beneficial to popularization and popularization of the surface plasmon biosensing technology.
In recent years, three-dimensional nano-patterned noble metal structure sensing designs and applications featuring low cost and small integration have shown better application prospects in the relevant fields. Compared with a sensor design based on a prism configuration, the characteristic size of the surface plasmon sensor adopting the three-dimensional patterned metal nano structure is close to or smaller than the wavelength of light, and a series of advantages are shown: (1) the complex optical coupling device which does not need angle control can directly excite the surface plasmon mode through vertical incidence, thereby being more convenient for the small integration of the structure; (2) the detection range is wider, and the sensing linearity is higher; (3) the structure design is more free, and the sensing mechanism is more flexible and diversified; (4) compatible with the existing imaging device and microfluidic technology, and provides possibility for the design of a multi-channel and high-flux biosensor; (5) the defect of low sensitivity of a prism type surface plasmon sensor in detecting small biological molecules is overcome, and the application range of biosensing is effectively expanded; (6) the applicable spectrum range is wider, and the ultraviolet light can be expanded to near infrared and even middle and far infrared wave bands. The surface plasmon sensor based on three-dimensional metal nanostructure electromagnetic coupling has the characteristics of miniaturization and integration, and therefore has a great commercial advantage compared with a prism coupling structure.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a high-flux multi-target microfluidic biochip based on a three-dimensional plasmon metamaterial, and solves some technical problems provided by the background technology. The method can overcome the defects of high price, complex light path, complex operation, limited size of detected molecules, large volume of detection equipment and the like of the first generation of plasmon biochip, and is more beneficial to popularization and generalization of the surface plasmon biosensing technology.
In order to achieve the purpose, the invention is realized by the following technical scheme: high flux many targets microflow biochip based on three-dimensional plasmon super structure material includes: a biochip; the biochip includes: the device comprises a plastic cover plate, a three-dimensional plasmon metamaterial sensing device, a microfluidic bottom plate and a plurality of biological reagents; the plastic cover plate, the three-dimensional plasmon metamaterial sensing device and the microfluidic bottom plate are compounded from top to bottom, wherein the plastic cover plate is adhered to the microfluidic bottom plate by an adhesive, and the three-dimensional plasmon metamaterial sensing device is embedded in a pit of a detection area of the microfluidic bottom plate; the biological reagents include, but are not limited to, 11-mercaptoundecanoic acid solution MUA, ethyldimethylamine propyl carbodiimide EDC, N-hydroxysuccinimide NHS, bovine serum albumin BSA, ligand solution, and analyte solution;
preferably, the plastic cover plate is circular, the radius is 4cm, the thickness is 2mm, 24 circular sample introduction through holes with the radius of 500 mu m are uniformly distributed in an array mode at the position 1cm away from the circle center, and the sample introduction through holes are matched with the microfluidic bottom plate;
preferably, the three-dimensional plasmon super-structure material sensing device is prepared by adopting a nano processing technology and is formed by compounding a flexible polycarbonate substrate, a chromium film and a gold film from bottom to top; three-dimensional quasi-nano columns are uniformly distributed on the flexible polycarbonate substrate in an array mode, the peripheries of the upper parts of the quasi-nano columns are rounded, and the top ends of the quasi-nano columns are flush;
preferably, the preparation process of the three-dimensional plasmon super-structure material sensing device comprises the following steps of firstly, carrying out hot stamping on a flexible polycarbonate substrate by adopting nano stamping; then, etching treatment is carried out by adopting an inductive coupling plasma etching machine; thirdly, plating a chromium film and a gold film by adopting an electron beam evaporation plating instrument; finally, it was cut into circular devices with a radius of 2 mm.
Preferably, the flexible polycarbonate substrate is in a periodic array nano-pore array structure, the diameter of the pores is 250nm, the depth of the pores is 200nm, and the period is 500 nm.
Preferably, the hot stamping time is 8min, the pressure is 40bar, and the temperature is 150 ℃.
Preferably, the etching treatment conditions are 8Pa, 40W of radio frequency power, 50sccm of oxygen flow and 20 ℃ of temperature.
Preferably, the thickness of the chromium film is 10nm, and the thickness of the gold film is 180 nm.
Preferably, the radius of the microfluidic bottom plate is 4cm, the thickness of the microfluidic bottom plate is 5mm, a clamping groove is arranged at the center of the circle, and 24 circular sample injection columns 302 with the radius of 500 micrometers are uniformly distributed in an array mode at the position 1cm away from the center of the circle; 24 circular detection areas with the radius of 2mm are uniformly distributed at the position 2cm away from the circle center in an array manner; 24 circular waste liquid areas with the radius of 4mm are evenly distributed in an array mode at the position 3.2cm away from the circle center, and the circular sample injection column, the circular detection area and the circular waste liquid areas are connected through a linear micro-flow channel with the width of 100 microns.
The method for measuring the molecular concentration and the dynamics of the high-flux multi-target microfluidic biochip based on the three-dimensional plasmon metamaterial comprises the following steps:
the method comprises the following steps: forming a self-assembled film on the surface of a biochip, activating, and placing the three-dimensional plasmon super-structure material sensing device in 10 mmol/L11-mercaptoundecanoic acid solution MUA at the temperature of 2-8 ℃ for 12 hours to form the MUA self-assembled film; and activating carboxyl by MUA at 2-8 ℃ by adopting 400mmol/L ethyl dimethyl amine propyl carbodiimide EDC and 100mmol/L N-hydroxysuccinimide NHS;
step two: fixing a ligand, namely putting the sensing device in the step one into a ligand solution, soaking for 2-4 hours at room temperature of 37 ℃ to enable the ligand to be combined with the MUA of the 11-mercaptoundecanoic acid solution on the surface, so as to fix the ligand;
step three: blocking the redundant carboxyl, adding bovine serum albumin BSA with the concentration of 50 mug/mL into the sensing device in the second step, wherein the bovine serum albumin BSA can be combined with the 11-mercaptoundecanoic acid solution MUA which is not combined with the ligand on the sensing device, so as to block the redundant carboxyl;
step four: adding an analyte solution to measure the molecular dynamics process, adding standard analyte solutions with different concentrations to perform determination, and representing the binding rate of the ligand and the analyte through the real-time offset of the spectrum;
step five: in the regeneration process, the chip can be regenerated in an acid environment, so that the repeated utilization is realized, and the cost is saved.
Preferably, 24 circular detection areas are used for carrying out synchronous real-time detection on different kinds of protein markers. Such as tumor markers carcinoembryonic antigen (CEA), carbohydrate antigen 199(CA199), carbohydrate antigen 125(CA125), carbohydrate antigen 153(CA153), alpha-fetoprotein (AFP), prostate cancer specific antigen (PSA); and detecting COVID-19 immunoglobulin IgM and COVID-19 immunoglobulin IgG.
Advantageous effects
The invention provides a high-flux multi-target microfluidic biochip based on a three-dimensional plasmon metamaterial. Compared with the prior art, the method has the following beneficial effects: the high-flux multi-target microfluidic biochip based on the three-dimensional plasmon metamaterial can provide 24-channel high-flux multi-marker parallel real-time detection, overcomes the defects that a first-generation plasmon biochip is high in price, complex in light path, complex in operation, limited in detection molecule size, large in detection equipment volume and the like, and is more beneficial to popularization and application of the surface plasmon biosensing technology.
Drawings
FIG. 1 is a schematic three-dimensional exploded view of a high-throughput biochip of the present invention;
FIG. 2 is a three-dimensional structure diagram of the high-throughput biochip of the present invention;
FIG. 3 is a scanning electron microscope side view of a three-dimensional plasmon metamaterial sensing device according to the present invention;
FIG. 4 is a schematic diagram of an external system framework equipped with a high-throughput biochip according to the present invention;
FIG. 5 is a schematic cross-sectional view of a molecular dynamics detection process of the plasmon three-dimensional excimer metamaterial sensing device.
In the figure: the device comprises a plastic cover plate 1, a sample introduction through hole 101, a three-dimensional plasmon metamaterial sensing device 2, a microfluidic baseboard 3, a flexible polycarbonate substrate 201, a chromium film 202, a gold film 203, a clamping groove 301, a circular detection area 303, a circular waste liquid area 304 and a linear microfluidic channel 305.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1-5, the method includes: a biochip; the biochip includes: the device comprises a plastic cover plate 1, a three-dimensional plasmon metamaterial sensing device 2, a microfluidic baseplate 3 and a plurality of biological reagents; the plastic cover plate 1, the three-dimensional plasmon metamaterial sensing device 2 and the microfluidic bottom plate 3 are compounded from top to bottom, wherein the plastic cover plate 1 is adhered to the microfluidic bottom plate 3 through an adhesive, and the three-dimensional plasmon metamaterial sensing device 2 is embedded in a pit of a detection area of the microfluidic bottom plate 3; the biological reagents include, but are not limited to, 11-mercaptoundecanoic acid solution MUA, ethyldimethylamine propyl carbodiimide EDC, N-hydroxysuccinimide NHS, bovine serum albumin BSA, ligand solution, and analyte solution;
furthermore, the plastic cover plate 1 is circular, the radius is 4cm, the thickness is 2mm, 24 circular sample feeding through holes 101 with the radius of 500 mu m are evenly distributed in an array mode at the position 1cm away from the circle center, and the sample feeding through holes 101 are matched with the micro-fluidic bottom plate 3;
further, the three-dimensional plasmon super-structure material sensing device 2 is prepared by adopting a nano processing technology and is formed by compounding a flexible polycarbonate substrate 201, a chromium film 202 and a gold film 203 from bottom to top; three-dimensional quasi-nano columns are uniformly distributed on the flexible polycarbonate substrate 201 in an array mode, the peripheries of the upper parts of the quasi-nano columns are rounded, and the top ends of the quasi-nano columns are flush;
further, the three-dimensional plasmon super-structure material sensing device 2 is prepared by the following steps of firstly, performing hot stamping on a flexible polycarbonate substrate 201 by adopting nano stamping; then, etching treatment is carried out by adopting an inductive coupling plasma etching machine; thirdly, plating a chromium film 202 and a gold film 203 by using an electron beam evaporation plating instrument; finally, it was cut into circular devices with a radius of 2 mm.
Further, the flexible polycarbonate substrate 201 is structured as a periodic array nano-pore array, the diameter of the pores is 250nm, the depth of the pores is 200nm, and the period is 500 nm.
Further, the hot stamping time is 8min, the pressure is 40bar, and the temperature is 150 ℃.
Further, the etching treatment conditions are that the pressure is 8Pa, the radio frequency power is 40W, the oxygen flow is 50sccm, and the temperature is 20 ℃.
Further, the thickness of the chromium film 202 is 10nm, and the thickness of the gold film 203 is 180 nm.
Referring to fig. 3, it can be seen from a side view of the scanning electron microscope of the three-dimensional plasmon metamaterial sensing device 2 that the prepared sensing device has uniform size and good morphology.
The radius of the microfluidic bottom plate 3 is 4cm, the thickness is 5mm, a clamping groove 301 is arranged at the center of a circle, and 24 circular sample injection columns 302 with the radius of 500 mu m are uniformly distributed in an array mode at the position 1cm away from the center of the circle; 24 circular detection areas 303 with the radius of 2mm are uniformly distributed at the position 2cm away from the circle center in an array manner; 24 circular waste liquid areas 304 with the radius of 4mm are evenly distributed in an array mode at the position 3.2cm away from the circle center, and the circular sample injection column 302, the circular detection area 303 and the circular waste liquid areas 304 are connected through a linear micro-flow channel 305 with the width of 100 mu m.
Referring to fig. 4, the required reagent is injected by a micro pump, the micro pump can be connected to the biochip circular sample injection column 302 by a micro flow tube to form a seal, the injection rate of the micro pump is 200 μ L/min-2000 μ L/min, and the spectrometer integrated in the portable detector can collect, process and upload optical signals to the analysis software to obtain the measurement result.
The method for measuring the molecular concentration and the dynamics of the high-flux multi-target microfluidic biochip based on the three-dimensional plasmon metamaterial comprises the following steps:
referring to fig. 5, step one: forming a self-assembled film on the surface of a biochip, activating, and placing the three-dimensional plasmon super-structure material sensing device 2 in 10 mmol/L11-mercaptoundecanoic acid solution MUA at the temperature of 2-8 ℃ for 12 hours to form the MUA self-assembled film; and activating carboxyl by MUA at 2-8 ℃ by adopting 400mmol/L ethyl dimethyl amine propyl carbodiimide EDC and 100mmol/L N-hydroxysuccinimide NHS;
step two: fixing a ligand, namely putting the sensing device in the step one into a ligand solution, soaking for 2-4 hours at room temperature of 37 ℃ to enable the ligand to be combined with the MUA of the 11-mercaptoundecanoic acid solution on the surface, so as to fix the ligand;
step three: blocking the redundant carboxyl, adding bovine serum albumin BSA with the concentration of 50 mug/mL into the sensing device in the second step, wherein the bovine serum albumin BSA can be combined with the 11-mercaptoundecanoic acid solution MUA which is not combined with the ligand on the sensing device, so as to block the redundant carboxyl;
step four: adding an analyte solution to measure the molecular dynamics process, adding standard analyte solutions with different concentrations to perform determination, and representing the binding rate of the ligand and the analyte through the real-time offset of the spectrum;
step five: in the regeneration process, the chip can be regenerated in an acid environment, so that the repeated utilization is realized, and the cost is saved.
Further, 24 circular detection areas 303 are used for carrying out synchronous real-time detection on different kinds of protein markers. Such as tumor markers carcinoembryonic antigen (CEA), carbohydrate antigen 199(CA199), carbohydrate antigen 125(CA125), carbohydrate antigen 153(CA153), alpha-fetoprotein (AFP), prostate cancer specific antigen (PSA); and detecting COVID-19 immunoglobulin IgM and COVID-19 immunoglobulin IgG.
The invention provides a high-flux multi-target microfluidic biochip based on a three-dimensional plasmon metamaterial, can provide 24-channel high-flux multi-marker parallel real-time detection, solves the defects of high price, complex light path, complex operation, limited detection molecule size, huge detection equipment volume and the like of a first generation of plasmon biochip, and is more beneficial to popularization and popularization of a surface plasmon biosensing technology.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (8)
1. High flux many targets microflow biochip based on three-dimensional plasmon super structure material includes: a biochip; the biochip includes: the sensor comprises a plastic cover plate (1), a three-dimensional plasmon metamaterial sensing device (2), a microfluidic base plate (3) and a plurality of biological reagents; the micro-fluidic chip is characterized in that the plastic cover plate (1), the three-dimensional plasmon metamaterial sensing device (2) and the micro-fluidic base plate (3) are compounded from top to bottom, wherein the plastic cover plate (1) and the micro-fluidic base plate (3) are adhered by adopting an adhesive, and the three-dimensional plasmon metamaterial sensing device (2) is embedded in a pit of a detection area of the micro-fluidic base plate (3); the biological reagents include, but are not limited to, 11-mercaptoundecanoic acid solution MUA, ethyldimethylamine propyl carbodiimide EDC, N-hydroxysuccinimide NHS, bovine serum albumin BSA, ligand solution, and analyte solution;
the plastic cover plate (1) is circular, the radius is 4cm, the thickness is 2mm, 24 circular sample introduction through holes (101) with the radius of 500 mu m are uniformly distributed in an array mode at the position 1cm away from the circle center, and the sample introduction through holes (101) are matched with the microfluidic base plate (3);
the three-dimensional plasmon super-structure material sensing device (2) is prepared by adopting a nano processing technology and is formed by compounding a flexible polycarbonate substrate (201), a chromium film (202) and a gold film (203) from bottom to top; three-dimensional quasi-nano columns are uniformly distributed on the flexible polycarbonate substrate (201) in an array mode, the peripheries of the upper parts of the quasi-nano columns are rounded, and the top ends of the quasi-nano columns are flush;
the radius of the microfluidic bottom plate (3) is 4cm, the thickness is 5mm, a clamping groove (301) is arranged at the center of the circle, and 24 circular sample injection columns (302) with the radius of 500 mu m are uniformly distributed in an array mode at the position 1cm away from the center of the circle; 24 circular detection areas (303) with the radius of 2mm are uniformly distributed at the position 2cm away from the circle center in an array manner; 24 circular waste liquid areas (304) with the radius of 4mm are evenly distributed in an array mode at the position 3.2cm away from the circle center, and the circular sample injection column (302), the circular detection area (303) and the circular waste liquid areas (304) are connected through a linear micro-flow channel (305) with the width of 100 mu m.
2. The three-dimensional plasmon metamaterial-based high throughput multi-target microfluidic biochip of claim 1, wherein: the three-dimensional plasmon super-structure material sensing device (2) is prepared by the following steps of firstly, carrying out hot stamping on a flexible polycarbonate substrate (201) by adopting nano stamping; then, etching treatment is carried out by adopting an inductive coupling plasma etching machine; thirdly, plating a chromium film (202) and a gold film (203) by adopting an electron beam evaporation instrument; finally, it was cut into circular devices with a radius of 2 mm.
3. The three-dimensional plasmon metamaterial-based high throughput multi-target microfluidic biochip of claim 1, wherein: the flexible polycarbonate substrate (201) is in a periodic array nano-pore array, the diameter of each pore is 250nm, the depth of each pore is 200nm, and the period is 500 nm.
4. The three-dimensional plasmon metamaterial-based high throughput multi-target microfluidic biochip of claim 2, wherein: the hot stamping time is 8min, the pressure is 40bar, and the temperature is 150 ℃.
5. The three-dimensional plasmon metamaterial-based high throughput multi-target microfluidic biochip of claim 2, wherein: the etching treatment conditions comprise pressure of 8Pa, radio frequency power of 40W, oxygen flow of 50sccm and temperature of 20 ℃.
6. The three-dimensional plasmon metamaterial-based high throughput multi-target microfluidic biochip of claim 1, wherein: the thickness of the chromium film (202) is 10nm, and the thickness of the gold film (203) is 180 nm.
7. The three-dimensional plasmon metamaterial-based high throughput multi-target microfluidic biochip of claim 1, wherein: the method for measuring the molecular concentration and the kinetics of the biochip comprises the following steps:
the method comprises the following steps: forming a self-assembled film on the surface of a biochip, activating, and placing the three-dimensional plasmon super-structure material sensing device (2) in 10 mmol/L11-mercaptoundecanoic acid solution MUA at the temperature of 2-8 ℃ for 12 hours to form an MUA self-assembled film; and activating carboxyl by MUA at 2-8 ℃ by adopting 400mmol/L ethyl dimethyl amine propyl carbodiimide EDC and 100mmol/L N-hydroxysuccinimide NHS;
step two: fixing a ligand, namely putting the sensing device in the step one into a ligand solution, soaking for 2-4 hours at room temperature of 37 ℃ to enable the ligand to be combined with the MUA of the 11-mercaptoundecanoic acid solution on the surface, so as to fix the ligand;
step three: blocking the redundant carboxyl, adding bovine serum albumin BSA with the concentration of 50 mug/mL into the sensing device in the second step, wherein the bovine serum albumin BSA can be combined with the 11-mercaptoundecanoic acid solution MUA which is not combined with the ligand on the sensing device, so as to block the redundant carboxyl;
step four: adding an analyte solution to measure the molecular dynamics process, adding standard analyte solutions with different concentrations to perform determination, and representing the binding rate of the ligand and the analyte through the real-time offset of the spectrum;
step five: in the regeneration process, the chip can be regenerated in an acid environment, so that the repeated utilization is realized, and the cost is saved.
8. The three-dimensional plasmon metamaterial-based high throughput multi-target microfluidic biochip of claim 1, wherein: 24 circular detection areas (303) are used for carrying out synchronous real-time detection on different kinds of protein markers.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010784687.8A CN111889154A (en) | 2020-08-06 | 2020-08-06 | High-flux multi-target microfluidic biochip based on three-dimensional plasmon metamaterial |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010784687.8A CN111889154A (en) | 2020-08-06 | 2020-08-06 | High-flux multi-target microfluidic biochip based on three-dimensional plasmon metamaterial |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111889154A true CN111889154A (en) | 2020-11-06 |
Family
ID=73245998
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010784687.8A Pending CN111889154A (en) | 2020-08-06 | 2020-08-06 | High-flux multi-target microfluidic biochip based on three-dimensional plasmon metamaterial |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111889154A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112547147A (en) * | 2020-11-23 | 2021-03-26 | 武汉世纪康敏生物科技有限公司 | Immunodetection chip and preparation method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050196779A1 (en) * | 2003-01-08 | 2005-09-08 | Ho Winston Z. | Self-contained microfluidic biochip and apparatus |
| CN101297189A (en) * | 2005-10-26 | 2008-10-29 | 通用电气公司 | Methods and systems for delivery of fluidic samples to sensor arrays |
| CN104823049A (en) * | 2012-10-01 | 2015-08-05 | 普林斯顿大学理事会 | Microfluidic sensors with enhanced optical signals |
| CN110836872A (en) * | 2019-11-19 | 2020-02-25 | 厦门大学 | Flexible label-free nano convex super-surface structure and manufacturing and sensing methods thereof |
| CN111077319A (en) * | 2019-12-18 | 2020-04-28 | 华南农业大学 | Micro-fluidic chip immunodetection kit and detection method thereof |
| US20200284788A1 (en) * | 2015-08-19 | 2020-09-10 | University Of Cincinnati | Patterned Plasmonic Nanoparticle Arrays for Multiplexed, Microfluidic Biosensing Assays |
-
2020
- 2020-08-06 CN CN202010784687.8A patent/CN111889154A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050196779A1 (en) * | 2003-01-08 | 2005-09-08 | Ho Winston Z. | Self-contained microfluidic biochip and apparatus |
| CN101297189A (en) * | 2005-10-26 | 2008-10-29 | 通用电气公司 | Methods and systems for delivery of fluidic samples to sensor arrays |
| CN104823049A (en) * | 2012-10-01 | 2015-08-05 | 普林斯顿大学理事会 | Microfluidic sensors with enhanced optical signals |
| US20200284788A1 (en) * | 2015-08-19 | 2020-09-10 | University Of Cincinnati | Patterned Plasmonic Nanoparticle Arrays for Multiplexed, Microfluidic Biosensing Assays |
| CN110836872A (en) * | 2019-11-19 | 2020-02-25 | 厦门大学 | Flexible label-free nano convex super-surface structure and manufacturing and sensing methods thereof |
| CN111077319A (en) * | 2019-12-18 | 2020-04-28 | 华南农业大学 | Micro-fluidic chip immunodetection kit and detection method thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112547147A (en) * | 2020-11-23 | 2021-03-26 | 武汉世纪康敏生物科技有限公司 | Immunodetection chip and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Hinman et al. | Surface plasmon resonance: material and interface design for universal accessibility | |
| Oliverio et al. | Chemical functionalization of plasmonic surface biosensors: a tutorial review on issues, strategies, and costs | |
| Chang et al. | High-sensitivity detection of carbohydrate antigen 15-3 using a gold/zinc oxide thin film surface plasmon resonance-based biosensor | |
| Hosseini et al. | Advantages, disadvantages and modifications of conventional ELISA | |
| Bolduc et al. | Advances in surface plasmon resonance sensing with nanoparticles and thin films: nanomaterials, surface chemistry, and hybrid plasmonic techniques | |
| Wang et al. | A label-free multisensing immunosensor based on imaging ellipsometry | |
| Li et al. | Revisiting the surface sensitivity of nanoplasmonic biosensors | |
| JP5397577B2 (en) | Surface plasmon resonance sensor and chip for the sensor | |
| JP5488469B2 (en) | Optical sensor, manufacturing method thereof, and detection method using optical sensor | |
| TWI384214B (en) | Biological sensing device and its system | |
| US20120019834A1 (en) | Multiplexed interferometric detection system and method | |
| KR20080082976A (en) | Sub-micron surface plasmon resonance sensor systems | |
| He et al. | Patterned plasmonic nanoparticle arrays for microfluidic and multiplexed biological assays | |
| KR101102454B1 (en) | Biochip with Metal-Protein Grating Pattern and Method for Preparing Thereof | |
| KR100737689B1 (en) | A method for amplication of signals of surface plasmon resonance sensor | |
| KR101093203B1 (en) | Copper-Capped Nanoparticle Array Biochip Based on LSPR Optical Properties and Use Thereof | |
| US20200225419A1 (en) | Dual grating sensing system, dual grating sensor and detecting method thereof | |
| CN111889154A (en) | High-flux multi-target microfluidic biochip based on three-dimensional plasmon metamaterial | |
| Rastogi et al. | Analyte Co-localization at Electromagnetic Gap Hot-Spots for Highly Sensitive (Bio) molecular Detection by Plasmon Enhanced Spectroscopies | |
| Krishnamoorthy et al. | Electrokinetic lab-on-a-biochip for multi-ligand/multi-analyte biosensing | |
| JP4302735B2 (en) | Biochip manufacturing method, biochip, biochip analyzer, biochip analysis method | |
| JP2013511714A5 (en) | ||
| CN111879707B (en) | Sensor, system and method of gold nanoparticle and quantum dot composite structure | |
| US20090036327A1 (en) | Method for chemical treatment of porous silicon surface | |
| Du et al. | Postcolumn renewal of sensor surfaces for high-performance liquid chromatography− surface plasmon resonance detection |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20201106 |