A multiplexed integrated microfluidic micro-ring resonator biosensor chip and a detection method thereof

Abstract

The application relates to a multiplexed integrated microfluidic micro-ring resonator cavity biosensor chip, which comprises a micro-flow channel layer and a main chip layer, wherein the micro-flow channel layer is arranged above the main chip layer; the micro-flow channel layer comprises a plurality of single sample inlet and outlet channels, a sample collection area and a single total sample inlet and outlet channel; the plurality of single sample inlet and outlet channels are each provided with an independent reaction chamber, each reaction chamber contains a micro-ring structure unit arranged on the main chip layer; and the plurality of micro-ring structure units are gradientally staggered. The multiplexed integrated microfluidic micro-ring resonator cavity biosensor chip provided by the application is a detection chip which can realize the detection of ten analytes in the same sample and the detection of the same analyte in ten samples, has high integration and economic advantages, and meanwhile, the gradient staggered arrangement of the plurality of micro-ring structure units can avoid mutual interference and improve the detection accuracy.

Description

Technical Field

[0001] This invention relates to the field of microfluidic chip fabrication technology, specifically to a multiplexed integrated microfluidic microring resonant cavity biosensor chip and its detection method. Background Technology

[0002] Microfluidics is a technology that creates micron-scale environments for precisely controlling and driving the flow of tiny liquids. It enables the controlled flow of minute liquids through target areas at controlled speeds and directions. Microfluidics offers significant advantages in detection, including multi-channel operation, miniaturization, low cost, small sample sizes, ease of operation, and high integration, making it a powerful tool in biosensing.

[0003] In addition, microfluidic technology can complete all analytical processes. Sample preparation, separation and reaction stages can all be integrated on the same microfluidic chip, which has great potential for real-time detection.

[0004] Label-free bioassay is a detection technology that enables quantification or semi-quantification of analytes without labeling the target analyte, providing real-time information for the analyte.

[0005] Label-free detection can detect the dynamic information of interactions between analytes in real time, making it one of the most ideal choices for instant detection.

[0006] However, label-free bioassays typically rely on complex analytical instruments, making it difficult to achieve large-scale application and miniaturization of label-free bioassays in non-laboratory environments.

[0007] Micro-nano optics has developed rapidly over the past few decades, and the integration of silicon photonics technology with biosensors has greatly broadened the application of optical biosensors.

[0008] However, individual channels on the same chip are relatively independent, making it impossible to detect multiple markers or multiple samples simultaneously on a single chip. Summary of the Invention

[0009] Technical problems to be solved

[0010] This invention proposes a multiplexed integrated microfluidic microring resonant cavity biosensor chip, the main purpose of which is to enable the simultaneous detection of multiple analytes of the same sample or the simultaneous detection of the same analyte of multiple samples on a single chip.

[0011] Efficient and diversified biosensing detection of multiple biomarkers can effectively improve biosensing efficiency and reduce detection costs.

[0012] Technical solution

[0013] To achieve the objectives of this invention, the technical solution adopted is as follows: a multiplexed integrated microfluidic microring resonant cavity biosensor chip, comprising:

[0014] A microfluidic channel layer and a main chip layer, wherein the microfluidic channel layer is disposed above the main chip layer;

[0015] The microfluidic channel layer includes: multiple individual sample inlet / outlet channels, a sample collection area, and a single overall sample inlet / outlet channel;

[0016] Multiple individual sample inlet and outlet channels are equipped with independent reaction chambers, each containing a microring structure unit located in the main chip layer; these microring structure units are arranged in a gradient-interlaced pattern. This detection chip can achieve dual functions: detection of ten analytes in the same sample and detection of the same analyte in ten samples. Furthermore, the gradient-interlaced arrangement of the multiple microring structure units avoids mutual interference, improving detection accuracy.

[0017] Preferably, the microring structure unit adopts a structure that satisfies the microring resonance principle to generate a resonant spectrum.

[0018] Preferably, the microring structural unit adopts a single microring structure, a double microring structure, a concentric ring structure, or a racetrack ring structure, and the microring structural unit realizes a microring immune base through biofunctionalization.

[0019] Preferably, the width of a single individual sample inlet / outlet channel reaction chamber ranges from 50 to 500 μm, the height of a single total sample inlet / outlet channel reaction chamber ranges from 10 to 200 μm, and the width of a single total sample inlet / outlet channel ranges from 50 to 1000 μm.

[0020] Compared with the prior art, the beneficial effects of the present invention are:

[0021] The multiplexed integrated microfluidic microring resonant cavity biosensor chip proposed in this invention is a detection chip capable of dual functions: detection of ten analytes in the same sample and detection of the same analyte in ten samples. This chip boasts high integration and economic advantages. It can achieve complex functions such as ten channels flowing through different solutions separately and ten channels simultaneously flowing through the same solution. Furthermore, it can utilize a combination of two detection methods to achieve the dual functions of detecting ten analytes in the same sample and detecting the same analyte in ten samples, thus expanding the application of multiplexed integrated microfluidic microring resonant cavity biosensor chips. Simultaneously, the gradient-staggered arrangement of multiple microring structural units avoids mutual interference and improves detection accuracy. Attached Figure Description

[0022] Figure 1 This is a top view of the structure of the multiplexed integrated microfluidic microring resonant cavity biosensor chip of the present invention;

[0023] Figure 2 This is a three-dimensional view of the chip structure of the multiplexed integrated microfluidic microring resonant cavity biosensor chip of the present invention.

[0024] Figure 3 An exploded view of the chip structure of the multiplexed integrated microfluidic microring resonant cavity biosensor chip of the present invention.

[0025] Figure 4 This is a schematic diagram of the analytical flow of different samples modified by multiple units of the multiplexed integrated microfluidic microring resonant cavity biosensor chip of the present invention.

[0026] Figure 5 This is a schematic diagram of the analytical flow of multiple units of the same sample modified by the multiplexed integrated microfluidic microring resonant cavity biosensor chip of the present invention.

[0027] In the figure: 1. Microfluidic channel layer 101, Individual sample inlet / outlet channel 102, Sample collection area 103, Total sample inlet / outlet channel 2. Main chip layer 3. Micro-ring structure unit. Detailed Implementation

[0028] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0029] Example 1:

[0030] like Figures 1 to 5 As shown, the present invention provides a technical solution: a multiplexed integrated microfluidic microring resonant cavity biosensor chip, comprising: a microfluidic channel layer 1 and a main chip layer 2, wherein the microfluidic channel layer 1 is disposed above the main chip layer 2, and the microfluidic channel layer 1 includes: multiple individual sample inlet / outlet channels 101, a sample collection area 102 and a single total sample inlet / outlet channel 103, wherein the sample collection area 102 connects the multiple individual sample inlet / outlet channels 101 and the single total sample inlet / outlet channel 103, and this embodiment uses ten individual sample inlet / outlet channels 101.

[0031] Each of the ten separate sample entry and exit channels 101 is equipped with an independent reaction chamber. Each reaction chamber contains a micro-ring structure unit 3 located in the main chip layer 2. Multiple micro-ring structure units 3 are arranged in a gradient staggered manner to avoid mutual interference.

[0032] The ten micro-ring structural units 3 of the main chip layer 2 and the ten individual sample inlet and outlet channels 101 of the microfluidic channel layer 1 work together to achieve the modification of the same sample or different samples in the ten micro-rings.

[0033] Samples can be introduced through ten individual inlet / outlet channels 101, flow through the sample collection area 102, and then flow out through a single main inlet / outlet channel 103. In this case, different samples can be introduced into the ten individual inlet / outlet channels 101 and flow out through one outlet. Different samples can be in the ten individual inlet / outlet channels 101.

[0034] Samples can also be introduced through a single total inlet / outlet channel 103, flow through the sample collection area 102, and then flow out through ten individual inlet / outlet channels 101. In this case, the same sample is introduced into the single total inlet / outlet channel 103 and flows out through the ten individual inlet / outlet channels 101. The same sample is in the ten individual inlet / outlet channels 101, and the ten micro-loops can modify or capture the same sample.

[0035] The microring structural unit 3 adopts a structure that satisfies the microring resonance principle to generate a resonant spectrum. The microring structural unit 3 can adopt a single microring structure, a double microring structure, a concentric ring structure, or a racetrack ring structure. At the same time, the microring structural unit 3 realizes a microring immune base through biofunctionalization.

[0036] The width of a single individual sample inlet / outlet channel 101 reaction chamber ranges from 50 to 500 μm, the height of a single total sample inlet / outlet channel 103 reaction chamber ranges from 10 to 200 μm, and the width of a single total sample inlet / outlet channel 103 ranges from 50 to 1000 μm.

[0037] The microring resonant cavity biosensor is a type of refractive index-based sensor that detects light wavelengths (λ). res Within the ring, the following conditions are met:

[0038] Ln eff =mλ res

[0039] Where L is the circumference of the ring, and m is an integer, also called the modulus (m = 1, 2, 3, ...). The refractive index (n) near the microring structure. eff When the refractive index changes, the overall effective refractive index of the microring structure changes accordingly, and the resonant wavelength of the output spectrum in the straight waveguide will also change accordingly.

[0040] By establishing a calibration curve for the concentration of analytes based on the resonant wavelength change and corresponding concentration collected at the output end of the straight waveguide, quantitative detection of analytes with unknown concentrations can be performed.

[0041] Example 2:

[0042] S1: Obtain a sample to be tested, and inject the sample into a single total inlet / outlet channel 103;

[0043] S2: The sample to be tested flows through the sample collection area 102;

[0044] S3: The sample to be tested is diverted from the sample collection area 102 into multiple separate sample entry and exit channels 101. The sample to be tested passes through multiple micro-ring structure units 3 in sequence to complete sample capture.

[0045] S4: The waste liquid of the sample to be tested after flowing through the ten micro-ring units flows out from the ten channels through the ten separate inlet and outlet channels 101 and is discharged from the chip.

[0046] like Figures 1 to 4 As shown, the purpose of this embodiment is to detect ten analytes from the same sample. Further, to detect ten analytes from the same sample, it is necessary to first modify the antibodies corresponding to the ten analytes on ten microloop units respectively, and then simultaneously introduce the same sample into the ten microloop units.

[0047] A detection method for a multiplexed integrated microfluidic microring resonant cavity biosensor chip includes:

[0048] Among them, the following are adopted Figure 4 The microfluidic method involves introducing samples through ten individual sample inlet / outlet channels 101 of the chip, sequentially passing in antibody (PBS Buffer) containing 10 μg / mL and maintaining at room temperature for 1 hour;

[0049] Among them, the following are adopted Figure 5 The microfluidic method involves injecting samples into a single total inlet / outlet channel 103 of the microring chip, rinsing with PBS buffer, and maintaining at room temperature for 30 min with 1% BSA to seal the remaining free surface and prevent nonspecific adsorption.

[0050] After rinsing with PBS Buffer, the microcircular immune substrate was stored at 4°C for further use;

[0051] The immunoassay process uses, for example Figure 5 The microfluidic method involves injecting a single total sample inlet / outlet channel 103 of the chip, passing an antigen (PBS Buffer) solution into the microfluidic channel of the microring immune substrate, and incubating at room temperature for 30 minutes. After immune recognition, the sample is rinsed with PBS Buffer.

[0052] Example 3:

[0053] like Figures 1 to 5 As shown, the purpose of this embodiment is to detect the same analyte in ten samples. Further, to detect the same analyte in ten samples, it is necessary to first modify the antibody corresponding to the same analyte on ten microloop units simultaneously, and then pass ten samples into each of the ten microloop units.

[0054] A detection method for an integrated microfluidic microring resonant cavity biosensor chip for multiplexing, comprising:

[0055] S1: Acquire multiple samples to be tested, and sequentially inject the samples into the individual sample inlet / outlet channel 101;

[0056] In this embodiment, ten types of samples are used for testing.

[0057] S2: In multiple separate sample entry and exit channels 101, the sample to be tested passes through the micro-ring structure unit 3 to complete sample capture;

[0058] S3: After the sample to be tested completes the capture of the micro-ring structure unit 3, the sample to be tested flows through the sample collection area 102;

[0059] S4: The samples to be tested mixed in the sample collection area 102 flow out through a single total inlet / outlet channel 103 and are discharged from the chip.

[0060] Among them, the following are adopted Figure 5 The microfluidic method involves introducing a single total inlet / outlet channel 103 of the microring chip, sequentially introducing a homoantibody (PBS Buffer) containing 10 μg / mL and maintaining it at room temperature for 1 hour;

[0061] Among them, the following are adopted Figure 5 The microfluidic method involves injecting samples into a single total inlet / outlet channel 103 of the microring chip, rinsing with PBS buffer, and maintaining at room temperature for 30 min with 1% BSA to seal the remaining free surface and prevent nonspecific adsorption.

[0062] After rinsing with PBS Buffer, the microcircular immune substrate was stored at 4°C for further use;

[0063] The immunoassay process uses, for example Figure 4 The microfluidic method involves introducing ten individual sample inlet and outlet channels 101 of the microring chip, sequentially passing ten antigen (PBS Buffer) solutions into the microfluidic channels of the microring immune substrate, and incubating at room temperature for 30 minutes. After immune recognition, the sample is rinsed with PBS Buffer.

[0064] The embodiments disclosed herein are preferred embodiments, but are not limited thereto. Those skilled in the art can readily grasp the spirit of the present invention based on the above embodiments and make different extensions and variations, but as long as they do not depart from the spirit of the present invention, they are all within the protection scope of the present invention.

Claims

1. A multiplexed integrated microfluidic microring resonant cavity biosensor chip, characterized in that, include: A microfluidic channel layer and a main chip layer, wherein the microfluidic channel layer is disposed above the main chip layer; The microfluidic channel layer includes: multiple individual sample inlet and outlet channels, a sample collection area, and a single total sample inlet and outlet channel. One end of the sample collection area is connected to multiple parallel individual sample inlet and outlet channels, and the other end of the sample collection area is connected to the total sample inlet and outlet channel. Each of the multiple individual sample inlet and outlet channels is equipped with an independent reaction chamber, and each reaction chamber contains a microring structure unit set in the main chip layer; the multiple microring structure units are arranged in a gradient staggered manner.

2. The multiplexed integrated microfluidic microring resonant cavity biosensor chip according to claim 1, characterized in that, The microring structure unit adopts a structure that satisfies the microring resonance principle to generate a resonant spectrum.

3. The multiplexed integrated microfluidic microring resonant cavity biosensor chip according to claim 2, characterized in that, The microring structural unit adopts a single microring structure, a double microring structure, a concentric ring structure, or a racetrack ring structure. The microring structural unit realizes a microring immune base through biofunctionalization.

4. The multiplexed integrated microfluidic microring resonant cavity biosensor chip according to claim 1, wherein the width of the individual sample inlet / outlet channel reaction chamber is 50-500 μm, the height of the individual total sample inlet / outlet channel reaction chamber is 10-200 μm, and the width of the individual total sample inlet / outlet channel is 50-1000 μm.

5. A detection method for an integrated microfluidic microring resonant cavity biosensor chip with multiplexing as described in any one of claims 1 to 4, characterized in that, include: S1: Acquire multiple samples to be tested, and sequentially inject the samples into the test channel from separate inlet and outlet channels; S2: In multiple separate inlet and outlet channels, the sample to be tested passes through the micro-ring structure unit to complete sample capture; S3: After the sample to be tested completes the capture of the micro-ring structure unit, the sample to be tested flows through the sample collection area; S4: The samples to be tested, mixed in the sample collection area, flow out of the chip through a single total inlet / outlet channel.

6. The detection method of the multiplexed integrated microfluidic microring resonant cavity biosensor chip according to claim 5, characterized in that, Samples were introduced through multiple separate inlet and outlet channels, and PBS buffer containing 10 μg / mL antibody was sequentially introduced and kept at room temperature for 1 hour. Samples were introduced into a single total inlet / outlet channel of the microring chip, rinsed with PBS buffer, and kept at room temperature for 30 min with 1% BSA to seal the remaining free surface and prevent nonspecific adsorption. After rinsing with PBS Buffer, the microcircular immune substrate was stored at 4°C for further use; The immunoassay process employs a microfluidic method, with samples introduced into a single total inlet / outlet channel of the chip. PBS buffer solution containing the antigen is passed through the microfluidic channel of the microring immunoassay substrate and incubated at room temperature for 30 min. After immunorecognition, the sample is rinsed with PBS buffer.

7. A detection method for an integrated microfluidic microring resonant cavity biosensor chip with multiplexing as described in any one of claims 1 to 4, characterized in that, include: S1: Obtain a sample to be tested and introduce it through a single main inlet / outlet channel; S2: The sample to be tested flows through the sample collection area; S3: The sample to be tested is diverted from the sample collection area into multiple separate inlet and outlet channels. The sample to be tested passes through multiple micro-ring structure units in sequence to complete sample capture. S4: The waste liquid of the sample to be tested after flowing through the ten micro-ring units flows out from the ten channels through ten separate inlet and outlet channels and is discharged from the chip.

8. The detection method of the multiplexed integrated microfluidic microring resonant cavity biosensor chip according to claim 7, characterized in that, Samples were introduced into a single total inlet and outlet channel of the microring chip, and PBS buffer containing 10 μg / mL of the same antibody was sequentially introduced and kept at room temperature for 1 hour. Inject the sample through a single total inlet / outlet channel, rinse with PBS buffer, and incubate with 1% BSA at room temperature for 30 min to seal the remaining free surface and prevent nonspecific adsorption. After rinsing with PBS Buffer, the microcircular immune substrate was stored at 4°C for further use; The immunoassay process employs a microfluidic method, with samples introduced through ten individual inlet and outlet channels of the microring chip. Ten PBS buffer solutions containing antigens are sequentially passed through the microfluidic channels of the microring immunoassay substrate and incubated at room temperature for 30 minutes. After immunorecognition, the sample is rinsed with PBS buffer.

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