Array food allergen detection microfluidic chip, clamp and detection system

Abstract

This invention provides an array-type microfluidic chip, fixture, and detection system for detecting food allergens. The chip includes: a first microfluidic chip and a second microfluidic chip sharing a common substrate. The first microfluidic chip includes a first channel group disposed on the substrate, and the second microfluidic chip includes a second channel group disposed on the substrate. The first channel group includes at least two parallel longitudinal channels, one of which is used as a blank control, and the remaining longitudinal channels are used to modify target antibodies. The second channel group includes at least two parallel transverse channels, which are used for sample loading and fluorescent labeling. The transverse channels are vertically attached to the modified imprints of the longitudinal channels, and the intersections of the transverse and longitudinal channel modified imprints form array points.

Description

Technical Field

[0001] This invention relates to the field of allergen detection technology, specifically to an array-type microfluidic chip, fixture, and detection system for detecting food allergens. Background Technology

[0002] Food allergy is a pathological reaction that occurs after a patient comes into contact with a food allergen. It can damage multiple organs, including the respiratory, digestive, and skin systems, and in severe cases, can lead to anaphylactic shock or even death. Over the past few decades, the incidence of food allergies has been increasing year by year.

[0003] Therefore, developing rapid detection methods for food allergens remains a top priority. Summary of the Invention

[0004] The purpose of this invention is to provide a simple-to-operate, highly sensitive array-type microfluidic chip for detecting food allergens, and to provide a non-chemically covalently bonded encapsulation technology for the microfluidic chip to solve the problem of irreversible chemical covalent bonding in microfluidic chips. Furthermore, this invention also provides a method for converting the target substance in solution into an aerosol state to simulate the allergen aerosol generation mechanism during food processing or heating, thereby improving the sensitivity of allergen detection.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] The first aspect of this invention provides an array-type microfluidic chip for detecting food allergens, comprising: a first microfluidic chip and a second microfluidic chip sharing a common substrate. The first microfluidic chip includes a first channel group disposed on the substrate, and the second microfluidic chip includes a second channel group disposed on the substrate. The first channel group includes at least two parallel longitudinal channels, one of which is used as a blank control, and the remaining longitudinal channels are used to modify target antibodies. Each longitudinal channel has an inlet A and an outlet A. The second channel group includes at least two parallel transverse channels, which are used for sample loading and fluorescent labeling. The at least two transverse channels share an inlet B and an outlet B. The transverse channels are vertically attached to the modified imprints of the longitudinal channels, and the intersections of the transverse and longitudinal channels form array points. The two microfluidic chips are sequentially encapsulated within the substrate (i.e., the first microfluidic chip is encapsulated first, followed by the second microfluidic chip), and a detection area is formed at the channel intersections, constituting array points, enabling simultaneous high-throughput detection of several proteins. The channel width of the microfluidic chip can be at the micrometer level. By using an adaptive microfluidic chip fixture to package array-type microfluidic chips, good sealing performance of the microfluidic chips can be ensured without covalent bonding. Moreover, the packaging process is reversible, which facilitates the sequential assembly of different microfluidic chips on the same substrate.

[0007] Furthermore, the number of longitudinal channels is five, of which four longitudinal channels are used to modify one of four target antibodies respectively; wherein the four target antibodies include antibodies of OVA, OVO, LYS and TM.

[0008] Furthermore, the transverse flow channel is provided with a fishbone structure on the side away from the substrate to generate vortices uniformly in the internal fluid, thereby promoting the mixing of the target within the fluid.

[0009] Furthermore, there are four transverse flow channels, and the four transverse flow channels share one inlet B and one outlet B.

[0010] Furthermore, the substrate is made of glass; the first and second flow channel groups are respectively integrated onto PDMS. More specifically, the substrate is a glass slide.

[0011] A second aspect of this invention provides a clamp for holding an array-type food allergen detection microfluidic chip as described above. The clamp includes a loading slot, an adaptive clamping plate, and fixing pins. The top of the loading slot has an opening for placing the array-type food allergen detection microfluidic chip and the adaptive clamping plate. The sidewalls of the loading slot are provided with windows at different heights for accommodating the fixing pins. The adaptive clamping plate includes two clamping plates and several elastic members disposed between the clamping plates. The different windows are spaced at different distances from the bottom of the loading slot to better accommodate microfluidic chips of different thicknesses. The adaptive clamping plate can adaptively adjust the pressure applied to the microfluidic chip, ensuring good chip sealing while ensuring unobstructed microchannels and preventing channel collapse. It is understood that the loading slot also has corresponding longitudinal and transverse sample loading windows.

[0012] Furthermore, the bottom of the loading tank is provided with a longitudinal flow channel sample loading window and a transverse flow channel sample loading window; the longitudinal flow channel sample loading window is used to expose the inlet A and outlet A; the transverse flow channel sample loading window is used to expose the inlet B and outlet B.

[0013] A third aspect of the present invention provides a detection system comprising an array-type food allergen detection microfluidic chip as described above.

[0014] Furthermore, the device also includes an aerosol generation component and the aforementioned fixture, with the array-type food allergen detection microfluidic chip housed within the fixture. The aerosol generation component comprises a ceramic ultrasonic atomizing plate, an atomizing chamber cover, a transmission diaphragm, and an atomizing chamber arranged sequentially. The ceramic ultrasonic atomizing plate generates high-frequency vibrations, which are transmitted via a liquid medium to the transmission diaphragm and further to the test solution within the atomizing chamber, causing the test solution to vibrate and atomize, thus generating an aerosol. The atomizing chamber includes a lower solution storage area and an upper aerosol area, with the diameter of the solution storage area being smaller than the diameter of the aerosol area. The solution storage area is used for storing and atomizing the solution. The top of the aerosol area is provided with a carrier gas inlet and an aerosol outlet, allowing the carrier gas to enter through the inlet, fully contact the aerosol within the atomizing chamber, and ultimately carry the aerosol through the outlet into the detection device for reaction and analysis. By utilizing the aerosol generation component, the detection target state of the chip can be extended to an aerosol, improving the sensitivity and applicability of the detection target. This array-type chip can simultaneously detect multiple different allergens, better helping food allergy sufferers avoid potentially harmful allergens in food. Specifically, the medium liquid can be ultrapure water.

[0015] Furthermore, O-rings are provided on both sides of the transmission diaphragm.

[0016] Compared with the prior art, the technical solution provided by the present invention has at least the following advantages:

[0017] This invention provides an array-type microfluidic chip for food allergen detection, a matching adaptive microfluidic chip fixture, and a detection system. The chip maintains good sealing and pressure resistance without the need for covalent bonding. Simultaneously, the developed aerosol generation component converts the sample from a solution state into an aerosol state. Thanks to the concentration of the target in the aerosol, the sensitivity for target detection is significantly higher (approximately 8.5 times that of solution samples). Furthermore, this microfluidic chip is suitable for detecting allergens in solution or gas, offering a wider range of applications compared to traditional enzyme-linked immunosorbent assays (ELISA) methods that can only detect targets in solution. This invention provides a new approach for convenient and highly sensitive detection of food allergens. Attached Figure Description

[0018] One or more embodiments are illustrated by way of example with reference to the accompanying drawings. These illustrations do not constitute a limitation on the embodiments, and unless otherwise stated, the figures in the drawings are not to be limited by scale.

[0019] Figure 1A This is a schematic diagram of the structure of an array-type food allergen detection microfluidic chip provided by the present invention;

[0020] Figure 1B This invention provides a fluid behavior diagram of a microfluidic chip for detecting food allergens when the transverse flow channel has a fishbone structure.

[0021] Figure 1C This is a fluid behavior diagram of the transverse flow channel without a fishbone structure in an array-type food allergen detection microfluidic chip provided by the present invention;

[0022] Figure 2A An exploded view of an adaptive microfluidic chip fixture provided by the present invention;

[0023] Figure 2B A schematic diagram of the sample loading window of an adaptive microfluidic chip fixture provided by the present invention;

[0024] Figure 3 This is an exploded view of the sol-gel generation component in this invention;

[0025] Figure 4A This is a schematic diagram illustrating the working principle of an array-type microfluidic chip for detecting food allergens provided by the present invention.

[0026] Figure 4B Micrograph of the detection array of an array-type food allergen detection microfluidic chip provided by the present invention;

[0027] Figure 5A A schematic diagram of the structure of a detection system provided by the present invention;

[0028] Figure 5B An optimized aerosol flow rate diagram for a detection system provided by the present invention;

[0029] Figure 5C An optimization diagram of aerosol loading time for a detection system provided by the present invention;

[0030] Figure 5D A standard curve for the detection of OVA in aerosols was plotted under optimal conditions for the detection system provided by this invention.

[0031] Figure 5E This invention provides a characterization diagram of the relative fluorescence intensity consistency of 20 data points in an array of a detection system.

[0032] Figure 6 The image shows the multi-target detection results of a detection system provided by this invention.

[0033] In the figure, 100 - array-type food allergen detection microfluidic chip, 110 - first flow channel group, 111 - longitudinal flow channel, 120 - second flow channel group, 121 - transverse flow channel, 121a - fishbone structure, 115 - array point;

[0034] 200-Adaptive microfluidic chip fixture, 210-Loading slot, 211-Window, 212-Longitudinal flow channel sample loading window and transverse flow channel sample loading window, 220-Adaptive clamping plate, 221-Clamping plate, 222-Elastic element, 230-Fixing pin;

[0035] 300-Aerosol generating component, 310-Ceramic ultrasonic atomizing plate, 320-Atomizing chamber cover, 330-Transmission diaphragm, 340-Atomizing chamber, 341-Solution storage area, 342-Aerosol area, 350-O-ring seal. Detailed Implementation

[0036] As the background technology shows, developing rapid detection methods for food allergens remains a top priority.

[0037] This invention discloses a novel array-type microfluidic chip for detecting food allergens, capable of simultaneously and rapidly detecting multiple food allergens. The method employs a double-antibody sandwich approach. First, using the microfluidic chip channels, capture antibodies for different allergens are modified onto the chip substrate. Specifically, longitudinal channels for specifically capturing four food allergens in food extracts or aerosols are encapsulated on the substrate, leaving a modification imprint, and then the longitudinal channels are removed. Subsequently, transverse channels for labeling the captured food allergens are encapsulated on the substrate, used for sample loading and fluorescent labeling. This allows for highly sensitive quantitative detection of four corresponding food allergens in the test solution or aerosol: ovalbumin (OVA), ovomucoid (OVO), and lysozyme (LYS) from eggs, and tropomyosin (TM) from the crustacean mud crab.

[0038] This invention also discloses an adaptive fixture for microfluidic chip packaging, which has strong versatility and can provide a reliable and reversible microfluidic chip packaging solution without relying on chemical covalent bonding.

[0039] Furthermore, this invention also discloses a detection system, including a micro-device for generating aerosols from a solution, which can successfully convert the test protein solution in the solution into an aerosol state, simulating the allergen aerosol generation mechanism during food processing or heating. Simultaneously, because aerosols have a larger specific surface area, they can enhance the contact between food allergens and capturing antibodies, improving the sensitivity of allergen detection.

[0040] The present invention will now be described in detail with reference to specific embodiments.

[0041] Example 1: Array-type microfluidic chip for food allergen detection

[0042] The design principle of an array-type microfluidic chip for food allergen detection is as follows: Figure 1A As shown. The food allergen detection microfluidic chip of the present invention includes a first microfluidic chip and a second microfluidic chip. The first microfluidic chip includes a first channel group 110 disposed on a substrate, and the second microfluidic chip includes a second channel group 120 disposed on a substrate. The first channel group 110 includes at least two parallel longitudinal channels 111, one of which is used as a blank control, and the other longitudinal channels 111 are used to modify target antibodies. Each longitudinal channel 111 has an inlet A and an outlet A.

[0043] The chip features multiple (e.g., five in this example) independent longitudinal channels 111, each with a pair of inlets / outlets, facilitating the modification of various food allergen antibodies for simultaneous detection of multiple food allergens. The transverse channel 121 also has a pair of inlets / outlets; after fluid injection into the chip, it is divided into four portions, flowing into four parallel channels, allowing for four parallel experiments on each target simultaneously, ensuring accurate quantitative detection. In application, the longitudinal channel 111 is first encapsulated with a glass slide for modifying the capture antibody. After modification, the longitudinal channel 111 is removed, and then the transverse channel 121 is encapsulated with the same glass slide, followed by the injection of the test sample. The food allergen in the test sample reacts with the capture antibody located at the intersection of the transverse / longitudinal channels 111, resulting in an efficient and specific capture of the specific food allergen. After fluorescent labeling, the food allergen in the test sample can be quantitatively detected.

[0044] A fishbone structure 121a is placed at the top of the transverse flow channel 121, which can significantly mix the fluid and increase the probability of contact between the target in the fluid and the capture antibody modified on the glass slide, thereby improving the detection sensitivity. Simulations of the fluid distribution within the transverse flow channel 121 with and without the fishbone structure using COMSOL Multiphysics software revealed that the fluid within the flow channel with the fishbone structure 121a significantly generates eddies, such as... Figure 1B As shown. However, the fluid within the transverse flow channel 121 without a fishbone structure remains in a laminar flow state, with insignificant internal mixing, as... Figure 1C As shown. Therefore, it is confirmed that the fishbone structure 121a design is beneficial to improving detection sensitivity. It can be seen that with the fishbone structure 121a, the fluid distribution in the flow channel can generate eddies, which promotes the mixing of the target within the fluid; without the fishbone structure, the fluid distribution in the flow channel is laminar, and the mixing of the target within the fluid relies solely on diffusion, resulting in poor mixing effect.

[0045] Example 2 Adaptive Microfluidic Chip Fixture

[0046] Adaptive microfluidic chip fixtures such as Figure 2A As shown, it can be composed of a loading slot 210, an adaptive clamping plate 220, and a fixing pin 230. The inner wall dimensions of the chip loading slot 210 can be length × width × height = 76 × 26 × 30 mm, and according to the chip inlet / outlet design, a longitudinal flow channel sample loading window and a transverse flow channel sample loading window 212 corresponding to the first substrate can be provided at the bottom of the loading slot 210, such as... Figure 2BAs shown. Simultaneously, a pin window 211 is provided on the side wall of the loading slot 210. The height of the pin window 211 from the bottom can be adjusted according to the chip thickness, and two sets of pin windows 211 at different heights are provided to better adapt to the chip thickness. The adaptive clamping plate 220 is composed of upper and lower clamping plates 221 and springs in a sandwich configuration. The springs are key components for adjusting the clamping force, and springs of different strengths and sizes can be used according to the required airtightness. In application, the chip 100 and the adaptive clamping plate 220 are sequentially loaded and sealed with pins to complete the chip encapsulation. After application, the pins and the adaptive clamping plate 220 are sequentially removed, and the chip is taken out for subsequent analysis.

[0047] Example 3 Aerosol generation component in the detection system

[0048] Aerosol generation components such as Figure 3 As shown, it can be composed of a ceramic ultrasonic atomizing plate 310, an atomizing chamber cover 320, a transmission diaphragm 330, an O-ring seal 350, and an atomizing chamber 340. The atomizing chamber 340 is divided into a solution storage area 341 and an aerosol area 342 (not shown in the figure). The solution storage area 341 has a smaller diameter, which facilitates the storage and atomization of small amounts of solution; the diameter of the aerosol area 342 gradually increases from the bottom to the top, which facilitates aerosol diffusion. After the solution is atomized to form an aerosol, it is blown out by a carrier gas (such as nitrogen). The carrier gas inlet and outlet are located at the top of the aerosol area 342 to increase the travel distance of the carrier gas in the atomizing chamber 340 and improve the efficiency of aerosol loading.

[0049] Example 4: Array-based microfluidic chip for detecting food allergens detects ovalbumin in solution.

[0050] This invention uses ovalbumin (OVA) as an example to verify the conditions and performance of the array-type food allergen detection microfluidic chip of this invention in detecting food allergens. Its overall working principle is as follows: Figure 4A As shown, the longitudinal flow channel chip is first encapsulated to modify the capture antibody for food allergens. Each independent flow channel can be modified to capture an antibody for a specific food allergen. Then, the transverse flow channel chip is encapsulated for sample loading. The transverse flow channel chip has 4 parallel flow channels, which can perform 4 sets of parallel experiments on each protein target at the same time to verify the accuracy of the detection.

[0051] The specific implementation method is as follows: Take a clean glass slide, treat it with oxygen plasma, and then immerse it in a solution of (3-mercaptopropyl)-trimethoxysilane (MPTS, 4%, diluted with anhydrous ethanol) for 30 min at room temperature. Then, dry it with nitrogen gas and immerse it in a solution of 4-maleimide butyrate-N-hydroxysuccinimide ester (GMBS, 0.1 μM, dissolved in anhydrous ethanol) for 30 min at room temperature. Next, wash with anhydrous ethanol to remove unbound reagents and dry with nitrogen gas. Assemble the longitudinal flow channel chip and encapsulate it using the adaptive microfluidic chip fixture described in Example 2. Inject OVA-specific capture antibody (which can be rabbit-derived anti-OVA antibody, 1.5 μL / channel) and incubate at room temperature for 30 min. After incubation, wash with phosphate-buffered saline (PBST, containing 0.5% Tween 20, pH=7.2, 15 μL / channel). After removing the PBST from the channels, unencapsulate and remove the longitudinal flow channel chip. After drying the residual solution with nitrogen, the transverse flow channel chip is assembled and packaged using an adaptive microfluidic chip fixture. The inner wall of the flow channel is sealed with 5% BSA solution and washed with PBS. OVA standard solution is then loaded using a microinjection pump. After loading, unbound reagents are removed by washing with PBST, and a specific detection antibody for OVA (which can be a mouse-derived anti-OVA antibody) is injected. After incubation at room temperature for 30 min, unbound reagents are removed by washing with PBST, and finally, a fluorescently labeled secondary antibody (which can be a goat-derived anti-mouse IgG antibody, and the fluorescent labeling molecule can be Alexa Fluor 488 or any suitable fluorescent molecule) is injected. After incubation at room temperature for 30 min, unbound reagents are removed by washing with PBST, the solution in the flow channel is drained, the package is unsealed, the transverse flow channel chip is removed, and the surface salt ions are removed by washing with ultrapure water. After drying with nitrogen, imaging can be performed under a fluorescence microscope. Under optimal conditions, the limit of detection (LOD) for OVA in the detection solution is 66.67 ng / mL. After the fluorescence signals at each detection point in the array are digitized using image processing software, they can be used for quantitative analysis of the target, such as... Figure 4B As shown.

[0052] Example 5: Array-based microfluidic chip for detecting food allergens and detecting ovalbumin in aerosols.

[0053] Furthermore, since the array-type food allergen detection microfluidic chip of the present invention is closed and can withstand relatively high air pressure, it can be applied to the detection of food allergens in aerosols. In this embodiment, OVA is used as an example. First, the chip is assembled / modified according to the steps in Example 4, and then set aside. 500 μL of OVA solution is injected into the aerosol chamber described in Example 3 to generate an aerosol containing OVA molecules. Nitrogen gas is used as a carrier gas to blow the aerosol into the transverse flow channel chip, as shown below. Figure 5AAs shown. After sample loading, following the steps in Example 4, the transverse flow channel was cleaned with PBST, and the OVA to be tested was labeled sequentially with the detection antibody and fluorescent secondary antibody, followed by imaging and analysis. In this example, the aerosol flow rate and aerosol loading time were optimized, and the optimal aerosol loading flow rate was determined to be 30 mL / min. Figure 5B As shown, the optimal loading time is 30 minutes. Figure 5C As shown. Under the above optimal conditions, the LOD of OVA in the aerosol was plotted as 7.86 ng / mL, as shown. Figure 5D As shown, the detection limit is significantly lower than that of OVA in solution, meaning the sensitivity of food allergen detection in aerosol mode is approximately 8.5 times that of solution mode. The reasons are: (1) OVA in aerosol is dispersed in droplet form, resulting in a larger surface area, which is more conducive to the contact and reaction between OVA and the capture antibody; (2) the aerosol loading flow rate is higher, leading to better mixing of the fluid within the microchannel, which is more beneficial for the reaction between the target and the capture antibody. Furthermore, this embodiment also verified the consistency of relative fluorescence intensity among the 20 data points in the array, showing a relative fluorescence intensity dispersion coefficient of 6.9%, as... Figure 5E As shown, the relative fluorescence intensity of each data point in the array is highly consistent, ensuring the accuracy of the analysis of each data point in the array.

[0054] Example 6: Array-based microfluidic chip for detecting mixed food allergens in aerosols.

[0055] First, following the steps in Example 4, OVA capture antibodies were modified into the longitudinal chip. The inner wall of the microchannels was blocked with 5% BSA solution, and after washing with PBS, aerosols of OVA, TM, arginine kinase (AK), bovine serum albumin (BSA), DNA-RNA complex (S9.6), and streptavidin (SA) were loaded. After OVA detection antibody and fluorescent secondary antibody labeling, the fluorescence signals at each detection point in the chip were analyzed. The results showed that only the chip with OVA loaded had a high fluorescence signal, while other non-specific targets showed no signal, indicating that the chip has good target specificity. Figure 6 As shown in Figure A.

[0056] Subsequently, detection standard curves for OVO, LYS, and TM were plotted. Following the steps in Example 4, capture antibodies corresponding to OVO, LYS, and TM were modified into different chips, respectively. The inner walls of the microchannels were blocked with 5% BSA solution, and after washing with PBS, different concentrations of the target molecules were loaded to plot detection standard curves for the three targets. The results are as follows: Figure 6 As shown in Figures B to D. The calculated LOD values ​​for OVO, LYS, and TM in this scheme are 1.27, 4.25, and 0.31 ng / mL, respectively.

[0057] Finally, the chip in this scheme was used to detect mixed allergens in the aerosol. Following the steps in Example 4, capture antibodies corresponding to BSA, OVA, OVO, LYS, and TM were modified into channels 1-5 of the longitudinal flow channel, respectively. The inner walls of the microchannels were blocked with 5% BSA solution and washed with PBS for later use. OVA, OVO, LYS, and TM allergen solutions were taken and added to 5% BSA solution, with the final concentrations of OVA, OVO, LYS, and TM allergens being 40, 10, 80, and 20 ng / mL, respectively. After thorough mixing, 500 μL of the mixed solution was added to the aerosol chamber, and after generating an aerosol, it was purged into the chip with nitrogen. After washing the transverse flow channel with PBST, the target was labeled with detection antibodies and fluorescent secondary antibodies sequentially, and finally, imaging and analysis were performed. Because this chip has good target specificity, the capture antibodies modified in the longitudinal flow channel react only with their corresponding allergens, such as... Figure 6 As shown in Figure E, the fluorescence signal at the detection point can accurately reflect the concentration of the corresponding target in the aerosol, such as... Figure 6 As shown in Figure F. Ultimately, the detection concentrations of OVA, OVO, LYS, and TM were 41.14±3.37, 9.27±2.13, 78.52±4.87, and 18.91±4.11 ng / mL, respectively. This means the recoveries for the four allergens were 102.85%, 92.7%, 98.15%, and 94.55%, respectively, indicating that the chip has good accuracy in detecting food allergens.

[0058] In summary, the array-type food allergen detection microfluidic chip, fixture, and detection system provided by this invention have the following beneficial effects:

[0059] (1) Microfluidic chip:

[0060] The microfluidic chip consists of two intersecting channels, one horizontal and one vertical, which are sequentially encapsulated on a glass slide. The vertical channel is used to capture antibody (specific antibody against food allergens) for modification, while the horizontal channel is used for sample loading and fluorescent labeling. A fishbone-shaped structure can be designed at the top of the horizontal channel to facilitate the formation of eddies in the fluid (solution or aerosol) within the channel, enhancing fluid mixing and thus increasing the probability of contact between the target and the captured antibody, thereby improving the target detection sensitivity.

[0061] (2) Adaptive microfluidic chip fixture:

[0062] The adaptive microfluidic chip fixture features adjustable clamping force, facilitating better encapsulation of the microfluidic chip and glass slide. This ensures sufficient sealing of the microfluidic chip's channels while preventing channel collapse, facilitating the loading of solution or aerosol samples. Furthermore, the encapsulation is reversible, allowing for removal as needed.

[0063] (3) Aerosol generation component in the detection system:

[0064] The nebulization chamber is hollow and can be divided into a smaller solution storage area and an aerosol area whose diameter gradually increases from bottom to top. Using ultrasonic nebulization combined with nitrogen blowing, solution samples can be converted into aerosol samples. On the one hand, this method allows the aerosol to concentrate the target molecules, thereby improving the sensitivity of target detection; on the other hand, compared to traditional enzyme-linked immunosorbent assays (ELISA), which can only detect allergens in solution, the aerosol loading method can also detect gaseous samples, thus having a wider range of applications.

[0065] Those skilled in the art will understand that the above-described embodiments are specific examples of implementing this application, and in practical applications, various changes in form and detail may be made without departing from the spirit and scope of this application. Any person skilled in the art can make their own modifications and alterations without departing from the spirit and scope of this application; therefore, the scope of protection of this application should be determined by the scope defined in the claims.

Claims

1. An array-type microfluidic chip for detecting food allergens, characterized in that, include: A first microfluidic chip and a second microfluidic chip sharing a common substrate, wherein the first microfluidic chip includes a first channel group (110) disposed on the substrate, and the second microfluidic chip includes a second channel group (120) disposed on the substrate; The first flow channel group (110) includes at least two parallel longitudinal flow channels (111), one of which is used as a blank control and the other longitudinal flow channels (111) is used to modify the target antibody. Each longitudinal flow channel (111) has an inlet A and an outlet A. The second flow channel group (120) includes at least two parallel transverse flow channels (121), which are used for sample loading and fluorescent labeling. The at least two transverse flow channels (121) share an inlet B and an outlet B. The transverse flow channel (121) is vertically attached to the decorative mark of the longitudinal flow channel (111), and the intersection of the decorative mark of the transverse flow channel (121) and the longitudinal flow channel (111) forms an array of points (115). The first flow channel assembly (110) is used to encapsulate the substrate first and leave a modified imprint after specifically capturing food allergens in food extracts or aerosols; the second flow channel assembly (120) is used to encapsulate the substrate again after removing the first flow channel assembly (110), and to load samples and fluorescently labeled detection antibodies to label the captured food allergens.

2. The array-type food allergen detection microfluidic chip according to claim 1, characterized in that, The number of the longitudinal channels (111) is five, of which four of the longitudinal channels (111) are used to modify one of the four target antibodies respectively; Among them, the four target antibodies include ovalbumin, oval mucin, lysozyme, and tropomyosin derived from the crustacean mud crab.

3. The array-type food allergen detection microfluidic chip according to claim 1, characterized in that, The transverse flow channel (121) is provided with a fishbone structure (121a) on the side away from the substrate so that the internal fluid generates vortices uniformly, thereby promoting the mixing of the target in the fluid.

4. The array-type food allergen detection microfluidic chip according to claim 3, characterized in that, The number of transverse flow channels (121) is four, and the four transverse flow channels (121) share one inlet B and one outlet B.

5. The array-type food allergen detection microfluidic chip according to claim 1, characterized in that, The substrate is made of glass; the first flow channel group (110) and the second flow channel group (120) are respectively integrated on PDMS.

6. An adaptive microfluidic chip fixture, characterized in that, For clamping an array-type food allergen detection microfluidic chip as described in any one of claims 1 to 5; The fixture includes a loading slot (210), an adaptive clamping plate (220), and a fixing pin (230); The top of the loading slot (210) is provided with an opening for placing the array-type food allergen detection microfluidic chip and the adaptive clamp (220). The side wall of the loading slot (210) is provided with windows (211) at different heights for accommodating the fixing pin (230). The adaptive clamp (220) includes two clamps (221) and a plurality of elastic elements (222) disposed between the clamps (221).

7. The adaptive microfluidic chip fixture according to claim 6, characterized in that, The bottom of the loading tank (210) is provided with a longitudinal flow channel sample loading window and a transverse flow channel sample loading window (212); The longitudinal flow channel sample loading window is used to expose the inlet A and the outlet A; The transverse flow channel sample loading window is used to expose the inlet B and outlet B.

8. A detection system, characterized in that, Includes an array-type food allergen detection microfluidic chip as described in any one of claims 1 to 5.

9. The detection system according to claim 8, characterized in that, It also includes an aerosol generation component (300) and a fixture (200) as described in claim 7, wherein the array-type food allergen detection microfluidic chip (100) is disposed within the fixture (200); The aerosol generating component (300) includes a ceramic ultrasonic atomizing plate (310), an atomizing chamber cover (320), a transmission diaphragm (330), and an atomizing chamber (340) arranged sequentially. Among them, the ceramic ultrasonic atomizing sheet (310) is used to generate high-frequency vibration, which is transmitted to the transmission diaphragm (330) through the medium liquid, and further transmitted to the test solution in the atomizing chamber (340) to cause the test solution to vibrate and atomize, so as to generate aerosol; The atomizing chamber (340) includes a lower solution storage area (341) and an upper aerosol area (342), wherein the diameter of the solution storage area (341) is smaller than the diameter of the aerosol area (342). The solution storage area (341) is used for the storage and atomization of the solution; The top of the aerosol zone (342) is provided with a carrier gas inlet and an aerosol outlet, which are used to allow the carrier gas to enter through the inlet and fully contact the aerosol in the atomization chamber (340), and finally carry the aerosol from the outlet into the detection device for reaction and analysis.

10. The detection system according to claim 9, characterized in that, O-rings (350) are also provided on both sides of the transmission diaphragm (330).

Images