High-throughput multi-channel biomarker detection equipment and detection method

Abstract

The invention relates to high-throughput multichannel biomarker detection equipment and a detection method, which are used for solving the problems of insufficient detection efficiency, large-scale high-throughput detection, operation convenience, result consistency and the like of the existing biomarker detection equipment. The high-throughput multi-channel biomarker detection equipment comprises a photon chip, a photon chip mounting module, an optical fiber array coupling module, a visual positioning system, a light source module, a spectrograph detection module, an anti-crosstalk microfluidic module, a control and analysis module and a display screen, according to the device, through collaborative design of an array photon chip and multi-channel micro-fluidic control, a single chip can realize simultaneous detection of more than or equal to 100 channels at most, so that the detection quantity in unit time is greatly improved, and the device is suitable for multi-index combined screening and large sample rapid detection; the label-free detection mode based on the optical microcavity avoids complex labeling steps (such as enzyme, fluorescence and metal nanoparticles) in traditional immunoassay, and reagent dependence and operation cost are reduced.

Description

Technical Field

[0001] This invention relates to the fields of optical sensing, biological detection and microfluidics integration technology, specifically a high-throughput multi-channel biomarker detection device and method integrating photonic chips. Background Technology

[0002] With the advancement of precision medicine and the rapid growth in demand for multi-indicator joint screening of major diseases, biomarker detection technology is developing towards high throughput, automation, and miniaturization. Traditional detection methods, such as enzyme-linked immunosorbent assay (ELISA) and chemiluminescent immunoassay (CLIA), are relatively mature in terms of specificity, but they have many bottlenecks in the scenario of parallel detection of multiple targets: (1) Significant throughput limitation: Conventional ELISA only supports low-throughput detection, and can only detect a few target biomarkers each time. When facing dozens or even hundreds of targets, repeated operations are required, resulting in high reagent consumption and long detection time; (2) Low equipment integration: It is necessary to rely on multiple large instruments (such as microplate readers, plate washers, etc.) to complete the incubation, washing, signal reading and other processes. The size is huge and not suitable for portable or point-of-care testing (POCT); (3) High dependence on labeling: Most of them require enzyme or fluorescent labeling of antigens / antibodies, which is not only complicated to operate, but also prone to equipment errors due to different labeling efficiencies.

[0003] In recent years, label-free sensing technology based on optical resonant cavities has become a core direction for next-generation high-throughput biosensing technologies due to its high sensitivity, real-time monitoring capabilities, and chip-level miniaturization advantages. Typical structures such as Fabry-Perot (FP) microcavities, microring resonators (MRRs), and photonic crystal cavities can achieve sensitive identification of target molecules such as proteins, nucleic acids, and pathogens by detecting the response of the resonant wavelength to changes in the refractive index of the environment (the refractive index detection limit can reach 10). -6 -10 -8 (RIU). In addition, chip array integration has a natural advantage in realizing multi-channel parallel detection.

[0004] However, despite the excellent performance of individual optical sensing units, there are still challenges in building a high-throughput detection device that can be truly used in practical applications: (1) Complex chip coupling and installation: Traditional free-space optical paths or single-fiber alignment methods are difficult to adapt to multi-channel photonic sensing chips, resulting in low coupling efficiency, high alignment requirements, and poor repeatability. Existing coupling schemes require fine calibration of hundreds of channels one by one, which is complex and cannot guarantee the consistency of array-level coupling; (2) Insufficient microfluidic integration: Many optical sensing chips lack stable and reliable fluid injection devices, which cannot support high-throughput and high-repeatability experiments; (3) Lack of a unified platform for data acquisition and analysis: Multi-channel spectral reading, multi-target parallel identification and detection, and multivariate analysis have not yet formed standardized processing procedures; (4) Lack of commercial prototypes for complete equipment: Current research is mostly focused on single-point prototype verification, lacking an engineering platform that integrates high-density sensing chips, low-loss optical coupling, anti-crosstalk flow control, and signal processing and analysis.

[0005] Therefore, there is an urgent need to develop a new type of photonic chip detection device with complete functions and high-throughput detection capabilities. This device should be able to integrate the sensitivity advantages of microcavity sensors with multi-channel fluid control equipment, high-efficiency coupling structures, and intelligent data processing platforms to solve the shortcomings of existing technologies in terms of detection efficiency, ease of operation, and result consistency, and promote the practical application and large-scale deployment of photonic chips. Summary of the Invention

[0006] In view of the shortcomings of the existing technology, the purpose of this invention is to provide a high-throughput multi-channel biomarker detection device and detection method to solve the problems of existing biomarker detection devices in the background technology in terms of detection efficiency, ease of operation and consistency of results.

[0007] To achieve the above objectives, this invention proposes a high-throughput multi-channel biomarker detection device, comprising:

[0008] A photonic chip comprising multiple M×N (M≥10, N≥10) optical resonant cavity array structures, the photonic chip being used for highly sensitive detection of target molecules;

[0009] A photonic chip mounting module, wherein the photonic chip mounting module is used to mount photonic chips;

[0010] A fiber optic array coupling module is connected to a photonic chip and is used to couple test light from an external light source into the photonic chip.

[0011] A visual positioning system, comprising a high-precision optical camera, which acquires images of a photonic chip and a fiber optic array coupling module, thereby enabling real-time observation of the relative position and alignment status of the photonic chip and the fiber optic array coupling module.

[0012] A light source module is connected to a fiber array coupling module, and the light source module is used to provide a light source to the fiber array coupling module; the light source module is a tunable laser with a center wavelength of 1550nm or a broadband light source.

[0013] An optical signal detection module is connected to an optical fiber array coupling module. The optical signal detection module is used to receive the light transmitted by the optical fiber array coupling module and to perform spectral acquisition.

[0014] An anti-crosstalk microfluidic module is connected to a photonic chip. The anti-crosstalk microfluidic module is used to construct a fluid device with multiple independent channels on the surface of the photonic chip for independent transport and reaction of multiple target samples.

[0015] The control and analysis module is connected to the photonic chip, the photonic chip mounting module, the fiber array coupling module, the visual positioning system, the light source module, the optical signal detection module, and the anti-crosstalk microfluidic module. The control and analysis module is used to control the light source switching, spectrum acquisition, temperature adjustment, chip alignment, and fluid injection, and to process, analyze, and output the spectral signals.

[0016] The display screen is connected to the control and analysis module.

[0017] Preferably, the photonic chip is a microring resonator array, a Fabry-Perot microcavity array, or a photonic crystal microcavity array chip, which can achieve highly sensitive detection of target molecules;

[0018] Each optical resonant cavity of the photonic chip is modified with a specific biological probe, and each optical resonant cavity serves as an independent sensing unit.

[0019] The photonic chip is equipped with a grating coupler.

[0020] Preferably, the photonic chip mounting module includes a vacuum adsorption stage, a TEC thermoelectric cooler, a high-precision nanopiezoelectric displacement stage, and a temperature sensor, all of which are mounted and integrated on the vacuum adsorption stage.

[0021] The TEC thermoelectric cooler and the high-precision nanopiezoelectric displacement stage are connected to the temperature sensor. When the temperature sensor detects that the local temperature difference of the photonic chip is greater than 0.01℃, the TEC thermoelectric cooler and the high-precision nanopiezoelectric displacement stage are activated simultaneously to compensate and suppress thermal drift to less than 0.5pm.

[0022] Preferably, the fiber array coupling module includes a first port and a second port;

[0023] The first port is provided with an input terminal for light input and an output terminal for light output. The input terminal is connected to the light source module, and the output terminal is connected to the light signal detection module.

[0024] The second port is the chip coupling end. The second port corresponds to the fiber array end face, which is precision polished and has a tilt angle of 0°, 8°, 12°, 41°, 45° or other angles, and is permanently rigidly fixed.

[0025] The input and output signals are optically coupled through the same V-groove array.

[0026] Preferably, the light source module includes a laser and an optical switch, the laser and the optical switch are connected, and the input terminal of the first port is connected to the laser through the optical switch. The optical path includes 1×N optical switches, which are used to switch between different detection channels.

[0027] The laser is a tunable laser with a center wavelength of 1550nm or a broadband light source, and the light source output is time-division switched to different input fiber optic ports via an optical switch;

[0028] The laser has an FC or APC connector.

[0029] The optical switch can be a mechanical optical switch, a MEMS optical switch, a magneto-optical switch, or an electro-optical switch.

[0030] Preferably, the optical signal detection module includes a photodetector and a spectrometer, and the output of the first port is connected to the photodetector or the spectrometer.

[0031] The photodetector is a one-dimensional photodetector array; the spectrometer is a high-speed spectrometer.

[0032] Preferably, the anti-crosstalk microfluidic module is fabricated from PDMS using soft photolithography and includes multiple independent channels, each with a separate inlet and outlet. The channel chambers are precisely aligned with the preset functional regions of the photonic chip, which include, but are not limited to, channel 1 (anti-PSA antibody), channel 2 (anti-HER2 antibody), and channel 3 (anti-IL-6 antibody), with a positioning error ≤ ±5 μm.

[0033] Preferably, the control and analysis module includes a control module and an analysis module, and the control module and the analysis module are connected.

[0034] The control module includes buttons, a touch module, and an external communication interface;

[0035] The analysis module uses a Gaussian-Lorentz mixture model to fit and extract the resonance peaks;

[0036] The control module also includes a closed-loop feedback device, which uses the light intensity information collected by the photodetector to trigger a high-precision nanopiezoelectric displacement stage for alignment optimization, thereby enabling efficient optical coupling of the photonic chip.

[0037] The TEC thermoelectric cooler is a temperature control device with a temperature control stability of ±0.01℃, which can control the resonant wavelength shift caused by the temperature drift of the photonic chip to within 0.5pm.

[0038] Preferably, it also includes a pressure clamp for bonding the anti-crosstalk microfluidic module to the photonic chip.

[0039] To achieve the above objectives, this invention proposes a high-throughput multichannel biomarker detection method, providing the aforementioned detection equipment, and the steps include:

[0040] S1. Fix the photonic chip onto the vacuum adsorption stage of the photonic chip mounting module, and control the temperature of the TEC thermoelectric cooler to remain stable within a certain temperature range; for example, set the temperature within the range of 20-25℃.

[0041] S2. Connect the first port input of the fiber array coupling module to the tunable laser FC or APC connector through the input of the optical switch, and switch the optical signal to the optical path alignment channel through the optical switch.

[0042] S3. Connect the output end of the array fiber in the fiber optic connector of the fiber optic array coupling module to the photodetector array or spectrometer.

[0043] S4. Start the light source and switch the light switch to allow the excitation light to enter the first port input terminal, so that the excitation light is input into the photonic chip and outputs a transmission or reflection signal;

[0044] S5. By adjusting the high-precision nano-piezoelectric displacement stage in the photonic chip mounting module, the grating coupler on the photonic chip is aligned and coupled with the array end face (chip coupling end) of the fiber array coupling module.

[0045] S6. Use a pressure clamp to attach the anti-crosstalk microfluidic module to the photonic chip, and use an optical alignment microscope to align each channel with the preset functional area; where each channel is independently connected to one or more photonic chip sensing units of a target analyte.

[0046] S7. Inject the target biological sample into the channel of each anti-crosstalk microfluidic module, and the sample binds to the functionalized area on the chip surface; the optical switch synchronously switches to the channel of the anti-crosstalk microfluidic module (switching speed ≤ 0.1ms);

[0047] S8. Use the control and analysis software to control the spectrometer to acquire the spectral signal of each channel;

[0048] S9. Use the intelligent analysis module to process spectral signals;

[0049] S10 outputs a multi-channel resonant wavelength variation thermogram and a converted target analyte concentration report for further analysis or archiving.

[0050] Preferably, step S9 specifically includes:

[0051] S9.1 Spectral preprocessing, including background subtraction, filtering and normalization;

[0052] S9.2 Peak extraction: Extracting the resonant wavelength of the transmission spectrum through multi-peak fitting;

[0053] S9.3 Dynamic baseline correction: The environmental drift is collected through the non-functionalized reference channel, and the resonant wavelength shift of each sensing channel is deducted in real time.

[0054] S9.4 Temperature drift correction: The corrected resonant wavelength value is obtained through a functionalized reference channel containing a reference buffer solution that does not contain the target analyte.

[0055] S9.5 Concentration Conversion: Calculate the concentration of biomarkers based on the built-in standard curve (the number of target analytes shall not be less than 50).

[0056] Preferably, the channels in each anti-crosstalk microfluidic module characterize the binding of the target analyte by identifying the drift of the resonance peaks of microcavities such as FP optical resonators and microring resonators, with a wavelength resolution better than 0.001 nm.

[0057] Preferably, the detection cycle of each channel after sample injection is less than 3 minutes, and more than 50 target objects can be detected simultaneously in parallel.

[0058] Preferably, the photonic chip has a replaceable structure and supports a variety of functionalized probe chips to achieve joint detection of multiple markers such as proteins, nucleic acids, and small molecules.

[0059] Compared with the prior art, the present invention has the following advantages:

[0060] 1. High-throughput parallel detection capability: This device uses an array photonic chip and multi-channel microfluidics co-design to achieve simultaneous detection of up to ≥100 channels per chip, greatly increasing the detection volume per unit time. It is suitable for multi-indicator joint screening and rapid detection of large samples.

[0061] 2. Label-free and simplified operation: The label-free detection method based on optical resonant cavity avoids the complex labeling steps (such as enzymes, fluorescence, metal nanoparticles) in traditional immunoassays, reducing reagent dependence and operating costs.

[0062] 3. High detection sensitivity and accuracy: The high quality factor (Q value can reach 10) of the microcavity structure. 4 -10 5 This allows for the response to minute changes in refractive index (~10). -6 The RIU is highly sensitive, and when combined with a high-precision fitting algorithm, the wavelength extraction accuracy can reach ±0.5 pm, and the concentration detection sensitivity is better than 10 ng / mL.

[0063] 4. Stable optical path coupling and high repeatability: The array tilt angle V-groove and piezoelectric platform closed-loop control ensure automatic alignment of the optical fiber and the chip, solving the problems of cumbersome manual adjustment and poor stability of traditional coupling methods. The RSD of single chip repeatability detection is less than 3%.

[0064] 5. Strong anti-crosstalk capability: The microfluidic structure with independent liquid inlet / outlet design effectively avoids cross-contamination between channels and is suitable for mixed detection of various types of samples.

[0065] 6. High integration and compact size: The equipment has a desktop-level integrated structure with embedded modules such as laser, spectrometer, controller, and interface terminal, which makes it easy to promote and deploy in hospital laboratories, mobile testing vehicles, and primary clinics.

[0066] 7. Adaptable to multiple types of biomarker detection: The device is compatible with various detection tasks such as protein (e.g., cancer biomarkers), nucleic acid (e.g., viral nucleic acid fragments), and small molecule (e.g., toxins, pesticides), and has high versatility and expandability. Attached Figure Description

[0067] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0068] Figure 1 This is one of the device logic diagrams of Embodiment 1 of the present invention;

[0069] Figure 2This is the second device logic diagram of Embodiment 1 of the present invention;

[0070] Figure 3 This is a schematic diagram of the docking of a photonic chip of an optical resonant cavity unit array with an anti-crosstalk microfluidic module according to Embodiment 1 of the present invention;

[0071] Figure 4 This is a schematic diagram of the detection method flow according to Embodiment 2 of the present invention;

[0072] Figure 5 This is a schematic diagram of the photonic chip optical coupling feedback mechanism in Embodiment 1 of the present invention.

[0073] In the diagram: 1. Photonic chip; 2. Photonic chip mounting module; 3. Fiber optic array coupling module; 4. Visual positioning system; 5. Light source module; 6. Optical signal detection module; 7. Anti-crosstalk microfluidic module; 8. Control and analysis module; 9. Display screen. Detailed Implementation

[0074] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, 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. The components of the embodiments of the present invention described and shown in the accompanying drawings can generally be arranged and designed in various different configurations. Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0075] Example 1: As Figures 1-3 and Figure 5 As shown, this invention proposes a high-throughput multi-channel biomarker detection device, comprising:

[0076] Photonic chip 1 includes multiple 10×10 array optical resonant cavity units, forming 100 functionally independent detection units; photonic chip 1 is used for highly sensitive detection of target molecules;

[0077] Photonic chip mounting module 2, which is used to mount photonic chip 1;

[0078] Fiber optic array coupling module 3, which is connected to photonic chip 1, is used to couple test light from an external light source into photonic chip 1.

[0079] The visual positioning system 4 includes a high-precision optical camera. The visual positioning system 4 acquires images of the photonic chip 1 and the fiber array coupling module 3 through the high-precision optical camera, thereby observing the relative position and alignment status of the photonic chip 1 and the fiber array coupling module 3 in real time.

[0080] Light source module 5 is connected to fiber array coupling module 3 and is used to provide light source to fiber array coupling module 3.

[0081] The optical signal detection module 6 is connected to the fiber array coupling module 3. The optical signal detection module 6 is used to receive the light transmitted by the fiber array coupling module 3 and perform spectral acquisition.

[0082] Anti-crosstalk microfluidic module 7 is connected to photonic chip 1. The anti-crosstalk microfluidic module 7 is used to construct a fluid device with multiple independent channels on the surface of photonic chip 1 for independent transport and reaction of multiple target samples.

[0083] The control and analysis module 8 is connected to the photonic chip 1, the photonic chip mounting module 2, the fiber array coupling module 3, the visual positioning system 4, the light source module 5, the optical signal detection module 6, and the anti-crosstalk microfluidic module 7. The control and analysis module 8 is used to control the light source switching, spectrum acquisition, temperature adjustment, chip alignment, and fluid injection, and to process, analyze, and output the spectral signals.

[0084] Display screen 9 is connected to control and analysis module 8.

[0085] This embodiment Figure 1 The PC (i.e., computer) in the middle includes a control and analysis module 8 and a display screen 9.

[0086] The photonic chip 1 is a microring resonator array, a Fabry-Perot microcavity array, or a photonic crystal microcavity array chip, which can achieve highly sensitive detection of target molecules.

[0087] Each optical resonant cavity of the photonic chip 1 is modified with a specific biological probe, and each optical resonant cavity serves as an independent sensing unit. A grating coupler is disposed on the photonic chip 1.

[0088] This array design has the following characteristics:

[0089] The photonic chip 1 includes one or more optical resonant cavity unit arrays;

[0090] like Figure 3As shown, all resonant cavity units are connected in series through a single bus waveguide, with only a single input / output port, facilitating device integration; each optical resonant cavity is a high-Q FP resonant cavity (Q greater than 10). 4 Its cavity length and the gap between the reflectors are precisely designed and simulated to ensure stable resonance near the C-band;

[0091] Optical resonant cavity arrays support multi-channel operation based on wavelength division multiplexing. The resonant wavelengths of different channels can be dynamically tuned by changing the left and right periods of the cavity and the Bragg reflection structure, thereby supporting the simultaneous detection of multiple targets.

[0092] The photonic chip mounting module 2 includes a vacuum adsorption stage, a TEC thermoelectric cooler, a high-precision nanopiezoelectric displacement stage, and a temperature sensor. The TEC thermoelectric cooler, the high-precision nanopiezoelectric displacement stage, and the temperature sensor are all mounted and integrated on the vacuum adsorption stage.

[0093] The TEC thermoelectric cooler and the high-precision nanopiezoelectric displacement stage are connected to the temperature sensor. When the temperature sensor detects that the local temperature difference of the photonic chip 1 is greater than 0.01℃, the TEC thermoelectric cooler and the high-precision nanopiezoelectric displacement stage are activated simultaneously to compensate and suppress thermal drift to less than 0.5pm.

[0094] The photonic chip 1 is fixed on a vacuum adsorption stage with integrated TEC components, and the adsorption force is greater than 1 kPa to ensure stability;

[0095] The TEC thermoelectric cooler uses feedback from an internal temperature sensor to determine the current and direction of operation, thereby stabilizing the temperature within a set range (e.g., 25℃ ± 0.01℃).

[0096] The entire platform is mounted on a high-precision nanopiezoelectric displacement stage, which can achieve sub-nanometer-level adjustment of XYZ and rotation direction to correct chip position offset caused by temperature drift or mechanical disturbance.

[0097] The fiber array coupling module 3 includes a first port and a second port;

[0098] The first port is provided with an input terminal for light input and an output terminal for light output. The input terminal is connected to the light source module 5, and the output terminal is connected to the light signal detection module 6.

[0099] The second port is the chip coupling end. The second port corresponds to the fiber array end face, which is precision polished and has a tilt angle of 0°, 8°, 12°, 41°, 45° or other angles, and is permanently rigidly fixed.

[0100] The input and output signals are optically coupled through the same V-groove array.

[0101] The light source module 5 includes a laser and an optical switch, which are connected together. The input terminal of the first port is connected to the laser via the optical switch. The optical path includes 1×N optical switches, which are used to switch between different detection channels.

[0102] The laser is a tunable laser or a broadband light source with a center wavelength of 1550nm. The laser output is time-division coupled to different input fiber ports via an optical switch.

[0103] The laser has an FC or APC connector.

[0104] The optical switch can be a mechanical optical switch, a MEMS optical switch, a magneto-optical switch, or an electro-optical switch.

[0105] like Figure 5 As shown, the input light uses a tunable narrow linewidth laser, which, together with an optical switch array, enables channel switching.

[0106] The optical input is coupled to the beveled polished structure of photonic chip 1 through an optical fiber array, with matching angles of approximately 0°, 8°, 12°, 41° or other angles to reduce coupling loss;

[0107] The output light is guided out by the chip and connected to a multi-channel photodetector array;

[0108] An optical signal detection module is set in the optical path to monitor the coupling efficiency in real time. If the loss is found to be greater than 5dB, a high-precision nano-piezoelectric displacement stage is automatically triggered for fine adjustment to maintain the coupling loss at ≤0.8dB.

[0109] The optical signal detection module 6 includes a photodetector and a spectrometer, and the output of the first port is connected to the photodetector or the spectrometer.

[0110] The photodetector is a one-dimensional photodetector array; the spectrometer is a high-speed spectrometer.

[0111] The anti-crosstalk microfluidic module 7 is fabricated from PDMS using soft photolithography, such as Figure 3 As shown, it contains multiple independent channels, each corresponding to a sensing unit. The dead volume of each channel is less than 0.5 μL, and each channel has a separate inlet and outlet, supporting parallel loading of samples or buffer solutions. The pressurized sealing structure ensures no cross-contamination and supports continuous detection or multiple elution. The channel chambers are precisely aligned with the preset functional areas of photonic chip 1. The preset functional areas of photonic chip 1 include, but are not limited to, anti-PSA antibody in channel 1, anti-HER2 antibody in channel 2, and anti-IL-6 antibody in channel 3, with a positioning error ≤ ±5 μm.

[0112] The control and analysis module 8 includes a control module and an analysis module, which are connected together.

[0113] The control module includes buttons, a touch module, and an external communication interface;

[0114] The analysis module uses a Gaussian-Lorentz mixture model to fit and extract the resonance peaks;

[0115] The control module also includes a closed-loop feedback device, which uses the light intensity information collected by the photodetector to trigger a high-precision nanopiezoelectric displacement stage for alignment optimization, thereby enabling efficient optical coupling of the photonic chip.

[0116] The TEC thermoelectric cooler is a temperature control device with a temperature control stability of ±0.01℃, which can control the resonant wavelength shift caused by the temperature drift of the photonic chip 1 to within 0.5pm.

[0117] The laser sequentially excites each channel via an optical switch;

[0118] A multi-channel photoelectric detection array acquires transmission signals to obtain a complete transmission spectrum;

[0119] The built-in Gaussian-Lorentz hybrid model (Voigt type) fitting algorithm extracts and tracks resonance peaks, and combines a reference channel to achieve automatic drift correction, ensuring accurate and reliable detection data.

[0120] This device can perform parallel detection of multiple proteins, nucleic acids, and tumor markers, and has advantages such as high throughput, high sensitivity, and small sample consumption. It is suitable for various scenarios such as medical screening, nutritional monitoring, and rapid on-site testing.

[0121] Example 2, as Figure 4 As shown, based on the technical solution and working principle of Embodiment 1, this invention proposes a high-throughput multi-channel biomarker detection method, providing the aforementioned detection equipment, and the steps include:

[0122] S1. Fix the photonic chip 1 onto the vacuum adsorption stage of the photonic chip mounting module 2, and control the temperature of the TEC thermoelectric cooler to remain stable at 20-25℃.

[0123] S2. Connect the first port input of the fiber array coupling module 3 to the tunable laser FC or APC connector through the input of the optical switch, and switch the optical signal to the optical path alignment channel through the optical switch.

[0124] S3. Connect the array fiber output end in the fiber connector of the fiber array coupling module 3 to the photodetector array or spectrometer.

[0125] S4. Start the light source and switch the light switch to allow the excitation light to enter the first port input terminal, so that the excitation light is input into the photonic chip 1 and outputs a transmission or reflection signal;

[0126] S5. By adjusting the high-precision nano-piezoelectric displacement stage in the photonic chip mounting module 2, the grating coupler on the photonic chip 1 is aligned and coupled with the array end face (chip coupling end) of the fiber array coupling module 3.

[0127] S6. Use a pressure clamp to attach the anti-crosstalk microfluidic module 7 to the photonic chip 1, and use an optical alignment microscope to align each channel with the preset functional area; wherein each channel is independently connected to one or more photonic chip 1 sensing units of a type of target analyte.

[0128] S7. Inject the target biological sample into the channel of each anti-crosstalk microfluidic module 7, and the sample binds to the functionalized area on the chip surface; the optical switch synchronously switches to the channel of the anti-crosstalk microfluidic module 7 (switching speed ≤ 0.1ms);

[0129] S8. Use the control and analysis software to control the spectrometer to acquire the spectral signal of each channel;

[0130] S9. Use the intelligent analysis module to process spectral signals;

[0131] S10 outputs a multi-channel resonant wavelength variation thermogram and a converted target analyte concentration report for further analysis or archiving.

[0132] Preferably, step S9 specifically includes:

[0133] S9.1 Spectral preprocessing, including background subtraction, filtering and normalization;

[0134] S9.2 Peak extraction: Extracting the resonant wavelength of the transmission spectrum through multi-peak fitting;

[0135] S9.3 Dynamic baseline correction: The environmental drift is collected through the non-functionalized reference channel, and the resonant wavelength shift of each sensing channel is deducted in real time.

[0136] S9.4 Temperature drift correction: The corrected resonant wavelength value is obtained through a functionalized reference channel containing a reference buffer solution that does not contain the target analyte.

[0137] S9.5 Concentration Conversion: Calculate biomarker concentrations based on the built-in standard curve (target analytes number no less than 50). Each channel in the anti-crosstalk microfluidic module 7 characterizes the binding of target analytes by identifying the drift of resonance peaks in optical resonators such as FP optical resonators and microring resonators, with a wavelength resolution better than 0.001 nm.

[0138] After sample injection, the detection cycle of each channel is less than 3 minutes, and it can detect more than 50 targets in parallel at the same time.

[0139] The photonic chip 1 has a replaceable structure and supports a variety of functionalized probe chips to achieve joint detection of multiple biomarkers such as proteins, nucleic acids, and small molecules.

[0140] Example 3: Equipment Configuration and Temperature-Piezoelectric Co-control Mechanism

[0141] This embodiment details the device configuration and temperature-pressure coordinated control mechanism for supporting the stable operation of the optical resonant cavity photonic chip 1, covering the following technical details:

[0142] 1. Module composition and linkage logic

[0143] (1) Photonic chip 1 is mounted on the photonic chip mounting module:

[0144] Vacuum adsorption platform: The chip is fixed by vacuum adsorption to ensure that the chip does not shift.

[0145] TEC thermoelectric cooler module: control chip temperature is 25.00±0.01℃;

[0146] High-precision nanopiezoelectric displacement stage: resolution ≤10nm, capable of XYZ and rotational direction adjustment to adjust the position of photonic chip 1;

[0147] Temperature and pressure linkage mechanism: When the temperature difference between the chip edge and center is detected to be greater than 0.01℃, the device simultaneously triggers the macro temperature adjustment of the TEC thermoelectric cooler and the Z-axis fine adjustment of the high-precision nano-piezoelectric displacement stage to compensate for the changes in the optical resonant cavity structure caused by the local temperature gradient. This mechanism controls the resonant peak displacement caused by temperature drift to less than 0.5pm, which can effectively solve the problem of local stress or thermal deformation that is difficult to handle by traditional temperature control.

[0148] (2) Optical coupling feedback module:

[0149] A fixed tilt angle (41°±1°) and grating end-face spacing (50±5μm) are maintained between the chip and the fiber array, according to... Figure 5 The optical coupling feedback mechanism in the photonic chip 1 enables coupling calibration. A real-time light intensity monitor monitors the emitted signal; if the loss exceeds 5dB, it triggers XYZ direction position compensation, operating independently of the temperature control device.

[0150] (3) Microfluidic coupling alignment:

[0151] The PDMS layer is pressed onto the chip surface using a pressure fixture. Real-time images are acquired using an optical microscope, and an image recognition module determines the alignment deviation between the flow channel and the optical resonant cavity array. The platform performs fine-tuning in the XY direction based on image feedback, ultimately achieving an alignment error of ≤±5μm.

[0152] 2. Dynamic stability control in the testing process

[0153] After the biological sample is injected, the device maintains detection stability through triple collaborative feedback:

[0154] Main temperature control: The TEC thermoelectric cooler controls the overall temperature to remain constant at 25.00℃;

[0155] Stress compensation: Temperature sensing → piezoelectric platform fine-tuning → suppressing resonant displacement caused by thermal expansion of the cavity;

[0156] Coupling maintenance: The chip displacement caused by fluid disturbance will be compensated in real time by the piezoelectric platform driven by light intensity feedback to ensure optical path consistency.

[0157] Example 4: Multi-channel marker detection process and software calibration equipment

[0158] This embodiment provides a biomarker multi-channel detection process and signal processing software device adapted to the optical resonant cavity array photonic chip 1, constructing a complete automatic analysis process of "spectral acquisition - data decoding - result output", and especially optimizing the resonance peak extraction, drift compensation and multi-channel data comparison analysis algorithms.

[0159] 1. Testing process design

[0160] The entire multi-channel detection process includes the following steps:

[0161] S1. Equipment initialization: After the control software starts, it self-checks the status of the temperature control, high-precision nano-piezoelectric displacement stage and light source module 5, and confirms that the chip installation and microfluidic bonding are completed.

[0162] S2. Light source and acquisition path calibration: Tune the laser scanning wavelength range (e.g., 1500-1580nm), determine the position of the optimal optical power and resonant wavelength through the reference channel, and automatically set the scanning center wavelength position and resolution of each channel.

[0163] S3. Sample loading: Various biological samples are injected in parallel via a multi-channel microfluidic module, with each channel loading different concentrations or types of target molecules.

[0164] S4. Resonance spectrum acquisition: The laser switches wavelengths in sequence according to the channel to excite the laser, and the transmission spectrum is synchronously acquired and saved in real time through the photoelectric detection array.

[0165] S5. Data Analysis and Drift Calibration: The equipment software performs equipment error corrections such as temperature drift and mechanical stress based on the reference channel. All target channel signals are normalized with reference to the corresponding background spectrum and reference spectrum.

[0166] S6. Resonance peak extraction and displacement calculation:

[0167] A mixture of Gaussian and Lorentz fitting algorithm was used to extract the wavelength position of the resonant peak channel by channel and compare it with the baseline data to calculate the peak redshift / blueshift value (unit: pm).

[0168] S7. Concentration Inversion and Quantitative Output:

[0169] By combining the standard curve model (pre-calibrated), the peak position shift is mapped to the target molecule concentration, and detection reports and alarm information (such as warnings of excessive concentration or low detection limit) are automatically generated.

[0170] 2. Software Architecture and Core Algorithm Module

[0171] The software for this device consists of a graphical user interface (GUI) and multiple algorithm kernels, supporting automated batch processing, data visualization, and result export. Specific modules include:

[0172] (1) Spectrum acquisition module:

[0173] Controls the tuned laser to perform step scanning and acquires transmission spectra from multiple channels, supporting custom configuration of wavelength range / step size.

[0174] (2) Resonance Peak Fitting and Tracking Module:

[0175] Accuracy is enhanced by employing Voigt-type (Gaussian-Lorentz hybrid) curve fitting;

[0176] Multi-peak detection and fitting confidence thresholds can be set;

[0177] Real-time monitoring of peak levels over time enables dynamic tracking.

[0178] (3) Reference channel drift correction module:

[0179] By setting a background baseline through a fixed channel, we can determine equipment-related drift (such as overall peak position shift caused by temperature) and make proportional corrections in each detection channel.

[0180] (4) Signal normalization and denoising module:

[0181] Methods such as moving average and wavelet transform are introduced to filter out random noise;

[0182] All channels are normalized to the same background signal reference, improving the accuracy of cross-channel comparison.

[0183] (5) Concentration Inversion Module:

[0184] By combining a pre-set standard curve library (the standard curves for each target analyte can be calibrated periodically), the shift values ​​are mapped to the target molecule concentrations;

[0185] Supports linear / nonlinear regression fitting of standard curves for different biomarkers;

[0186] Displays quantitative results and concentration error estimates (confidence intervals / error bars).

[0187] (6) Results Output and Recording Module:

[0188] Generate a CSV data report containing timestamps, channel numbers, sample numbers, resonance peak wavelengths, peak position changes, and concentration results;

[0189] Supports batch export of test results and spectra in PDF format;

[0190] Automatic warnings can be set for channels with abnormal results.

[0191] 3. Analysis of Implementation Results and Advantages

[0192] This detection process, combined with the photonic chip 1 structure and temperature-controlled piezoelectric device of the present invention, can achieve detection of 100 channels, with the minimum detection concentration of each channel reaching the pg / mL level;

[0193] The software equipment has fast, stable, and automated analysis capabilities, greatly reducing the need for manual intervention;

[0194] The testing process is highly standardized and suitable for various application scenarios such as high-throughput medical screening, biological research, and drug screening.

[0195] The equipment has good scalability and compatibility, and supports interfacing with existing laboratory LIMS equipment to achieve full-process digitalization.

[0196] The terms "upper," "lower," "front," "rear," "left," and "right" used above are for descriptive purposes only and should not be construed as indicating or implying relative importance. Unless otherwise specifically stated, the relative steps, numerical expressions, and values ​​of components and steps set forth in these embodiments do not limit the scope of the invention.

[0197] Of course, the above description is only a specific embodiment of the present invention and is not intended to limit the scope of the present invention. All equivalent changes or modifications made to the structure, features and principles described in the claims of the present invention should be included in the scope of the claims of the present invention.

[0198] The above-described embodiments are merely specific implementations of the present invention, used to illustrate the technical solutions of the present invention, and are not intended to limit it. The scope of protection of the present invention is not limited thereto. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that any person skilled in the art can still modify or easily conceive of changes to the technical solutions described in the foregoing embodiments within the technical scope disclosed in the present invention, or make equivalent substitutions for some of the technical features; and these modifications, changes, or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention, and should all be covered within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A high-throughput multi-channel biomarker detection device, characterized in that, include: A photonic chip (1) comprising multiple M×N optical resonant cavity array structures, the photonic chip (1) being used for highly sensitive detection of target molecules; Photonic chip mounting module (2), the photonic chip mounting module (2) is used to mount photonic chip (1); Fiber array coupling module (3), which is connected to photonic chip (1), is used to couple test light from an external light source into photonic chip (1); The visual positioning system (4) includes a high-precision optical camera. The visual positioning system (4) acquires images of the photonic chip (1) and the fiber array coupling module (3) through the high-precision optical camera, thereby observing the relative position and alignment status of the photonic chip (1) and the fiber array coupling module (3) in real time. The light source module (5) is connected to the fiber array coupling module (3). The light source module (5) is used to provide a light source to the fiber array coupling module (3). The light source module (5) is a tunable laser with a center wavelength of 1550nm or a broadband light source. Optical signal detection module (6), which is connected to fiber array coupling module (3), is used to receive the light transmitted by fiber array coupling module (3) and perform spectral acquisition; Anti-crosstalk microfluidic module (7) is connected to photonic chip (1). The anti-crosstalk microfluidic module (7) is used to construct a fluid system with multiple independent channels on the surface of photonic chip (1) for independent transport and reaction of multiple target samples. The control and analysis module (8) is connected to the photonic chip (1), the photonic chip mounting module (2), the fiber array coupling module (3), the visual positioning system (4), the light source module (5), the optical signal detection module (6), and the anti-crosstalk microfluidic module (7), respectively. The control and analysis module (8) is used to control the light source switching, spectrum acquisition, temperature adjustment, chip alignment, fluid injection, and to process, analyze and output the spectral signal. The display screen (9) is connected to the control and analysis module (8).

2. The high-throughput multi-channel biomarker detection device according to claim 1, characterized in that, The photonic chip (1) is a microring resonator array, a Fabry-Perot microcavity array, or a photonic crystal microcavity array chip, which can achieve highly sensitive detection of target molecules; Each optical resonant cavity of the photonic chip (1) is modified with a specific biological probe, and each optical resonant cavity serves as an independent sensing unit.

3. A high-throughput multi-channel biomarker detection device according to claim 1 or 2, characterized in that, The photonic chip mounting module (2) includes a vacuum adsorption stage, a TEC thermoelectric cooler, a high-precision nanopiezoelectric displacement stage and a temperature sensor. The TEC thermoelectric cooler, the high-precision nanopiezoelectric displacement stage and the temperature sensor are all mounted and integrated on the vacuum adsorption stage. The TEC thermoelectric cooler and the high-precision nanopiezoelectric displacement stage are connected to the temperature sensor respectively. When the temperature sensor detects that the local temperature difference of the photonic chip (1) is greater than 0.01℃, the TEC thermoelectric cooler and the high-precision nanopiezoelectric displacement stage are started simultaneously to compensate and suppress thermal drift to less than 0.5pm.

4. A high-throughput multi-channel biomarker detection device according to claim 1 or 2, characterized in that, The fiber array coupling module (3) includes a first port and a second port; The first port is provided with an input terminal for light input and an output terminal for light output. The input terminal is connected to the light source module (5), and the output terminal is connected to the light signal detection module (6). The second port is the chip coupling terminal.

5. A high-throughput multi-channel biomarker detection device according to claim 4, characterized in that, The light source module (5) includes a laser and an optical switch, which are connected together. The input terminal of the first port is connected to the laser through the optical switch.

6. A high-throughput multi-channel biomarker detection device according to claim 4, characterized in that, The optical signal detection module (6) includes a photodetector and a spectrometer, and the output of the first port is connected to the photodetector or the spectrometer.

7. A high-throughput multi-channel biomarker detection device according to claim 1, 2, 5 or 6, characterized in that, The anti-crosstalk microfluidic module (7) is made of PDMS by soft lithography and contains multiple independent channels, each with a separate inlet and outlet.

8. A high-throughput multi-channel biomarker detection device according to claim 1, 2, 5 or 6, characterized in that, The control and analysis module (8) includes a control module and an analysis module, which are connected together; The control module includes buttons, a touch module, and an external communication interface; The analysis module uses a Gaussian-Lorentz mixture model to fit and extract the resonance peaks; The control module also includes a closed-loop feedback system, which uses the light intensity information collected by the photodetector to trigger a high-precision nanopiezoelectric displacement stage for alignment optimization, thereby enabling efficient optical coupling of the photonic chip.

9. A high-throughput multi-channel biomarker detection method, characterized in that, Providing the detection device according to any one of claims 1-8, the steps include: S1. Fix the photonic chip (1) on the vacuum adsorption stage of the photonic chip mounting module (2) and control the temperature of the TEC thermoelectric cooler to remain stable within a certain temperature range. S2. Connect the first port input of the fiber array coupling module (3) to the tunable laser FC or APC connector through the input of the optical switch, and switch the optical signal to the optical path alignment channel through the optical switch. S3. Connect the array fiber output end of the fiber connector in the fiber array coupling module (3) to the photodetector array or spectrometer. S4. Start the light source and switch the light switch to enter the first port input terminal so that the excitation light is input into the photonic chip (1) and outputs a transmission or reflection signal. S5. By adjusting the high-precision nano-piezoelectric displacement stage in the photonic chip mounting module (2), the grating coupler on the photonic chip (1) is aligned and coupled with the array end face (chip coupling end) of the fiber array coupling module (3); S6. Use a pressure clamp to attach the anti-crosstalk microfluidic module (7) to the photonic chip (1), and use an optical alignment microscope to align each channel with the preset functional area; wherein each channel is independently connected to one or more photonic chip (1) sensing units of a type of target analyte; S7. Inject the target biological sample into the channel of each anti-crosstalk microfluidic module (7), and the sample binds to the functionalized area on the chip surface; the optical switch synchronously switches to the channel of the anti-crosstalk microfluidic module (7) (switching speed ≤ 0.1ms); S8. Use the control and analysis software to control the spectrometer to acquire the spectral signal of each channel; S9. Use the intelligent analysis module to process spectral signals; S10 outputs a multi-channel resonant wavelength variation thermogram and a converted target analyte concentration report for further analysis or archiving.

10. The high-throughput multi-channel biomarker detection method according to claim 9, characterized in that, Step S9 specifically includes: S9.1 Spectral preprocessing, including background subtraction, filtering and normalization; S9.2 Peak extraction: Extracting the resonant wavelength of the transmission spectrum through multi-peak fitting; S9.3 Dynamic baseline correction: The environmental drift is collected through the non-functionalized reference channel, and the resonant wavelength shift of each sensing channel is deducted in real time. S9.4 Temperature drift correction: The corrected resonant wavelength value is obtained through a functionalized reference channel containing a reference buffer solution that does not contain the target analyte. S9.5 Concentration Conversion: Calculate biomarker concentrations based on the built-in standard curve.

Images