A fluorescent labeling method and labeling system for IgLON5 antibody IgG detection

Abstract

The present invention discloses a fluorescent labeling system and method for detecting IgLON5 antibody IgG, belonging to the field of medical detection technology. The system includes a detection component, a detection unit, a mounting unit, and an electromagnetic excitation device, wherein the detection component is composed of a substrate and a signal expression layer, the signal expression layer having a metal film and a plurality of detection bumps, and the top is coated with an IgLON5-specific antigen as a capture agent. The mounting unit is used to fix the detection component and can control its relative movement under the detection unit to achieve high-precision detection on a region-by-region basis. The electromagnetic excitation device generates an alternating electromagnetic field through an induction coil, inducing a tiny electric field on the metal film to optimize the directional attachment and binding efficiency of the target analyte. The electromagnetic excitation device is provided with a first working state and a second working state, and dynamically adjusts the duration of the two states based on the data feedback from the real-time detection unit to optimize the distribution of the target analyte and the signal amplification effect.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical detection, and in particular relates to a fluorescent labeling method and a labeling system for detecting IgLON5 antibody IgG. Background Art

[0002] In the field of immunoassays and molecular diagnosis, efficient and accurate detection of antibodies is of great significance for the early diagnosis of diseases and the formulation of treatment plans. As an important marker related to autoimmune diseases of the nervous system, the accuracy and sensitivity of IgLON5 antibody IgG detection directly affect the effectiveness of clinical diagnosis. However, traditional antibody detection methods often have problems such as insufficient sensitivity, high background noise and long detection time, which make it difficult to meet the needs of efficient detection in complex sample environments. In addition, after the fluorescent marker binds to the targeted antibody, the residual unbound marker may interfere with the detection signal and affect the accuracy of the results. Therefore, how to effectively remove non-specifically bound fluorescent markers during the detection process and achieve efficient capture and directional attachment of targeted analytes at the microscopic scale has become a problem that needs to be solved urgently in current technology.

[0003] According to the disclosed technical solutions, the technical solution with publication number CN102507958A proposes a method for detecting IgG content. By preparing standard solutions of different concentrations and drawing regression test curves, it can achieve rapid detection of multiple concentrations of IgG. The technical solution with announcement number UA26767U proposes an IgG immunoenzyme test system, proposing the preparation of a monoclonal antibody with a high affinity constant, culture fluid titer, and peroxidase conjugate titer to improve the detection accuracy of IgG. The technical solution with publication number US20010018220A1 proposes an IgG antibody detection method. After preparing the sample with a dehydrated anti-IgG complex, the color of the detection reagent is used to achieve rapid detection of IgG antibodies.

[0004] The above technical solutions all propose a variety of technical solutions for realizing and optimizing IgG antibody detection. With the deepening of research in related fields, more efficient IgG antibody detection methods and supporting detection systems can be proposed.

[0005] The foregoing discussion of the background art is intended only to facilitate an understanding of the present invention. This discussion does not acknowledge or admit that any of the material referred to is part of the common general knowledge. Summary of the Invention

[0006] The present invention discloses a fluorescent labeling system and method for detecting IgLON5 antibodies (IgG), belonging to the field of medical detection technology. The system comprises a detection component, a detection unit, a mounting unit, and an electromagnetic excitation device. The detection component comprises a substrate and a signal expression layer, wherein the signal expression layer comprises a metal film and a plurality of detection bumps, and an IgLON5-specific antigen is coated on top as a capture agent. The mounting unit is used to secure the detection component and control its relative movement under the detection unit to achieve high-precision detection on a region-by-region basis. The electromagnetic excitation device generates an alternating electromagnetic field through an induction coil, inducing a microscopic electric field on the metal film to optimize the directional attachment and binding efficiency of the target analyte. The electromagnetic excitation device is provided with a first operating state and a second operating state, and dynamically adjusts the duration of the two states based on data feedback from the real-time detection unit to optimize the distribution of the target analyte and the signal amplification effect.

[0007] The present invention adopts the following technical solution: a fluorescent labeling system for detecting IgLON5 antibody IgG, the fluorescent labeling system comprising:

[0008] The detection component is configured to construct an environment on the surface of the detection component for specific binding of the IgLON5 antibody and the marker with fluorescent properties;

[0009] a detection unit configured to capture an image of the fluorescent signal generated from the surface of the detection component and convert the image into digital image data;

[0010] a mounting unit, configured to fix the detection assembly and enable the detection assembly and a detection element of the detection unit to move relative to each other, so as to allow the detection element to detect various areas of the detection assembly;

[0011] an analysis unit, configured to process and analyze the image data and output a detection result;

[0012] During pretreatment of the fluorescent marker, the fluorescent marker is chemically modified to impart a weak charge, so that the target marker formed after the fluorescent marker specifically binds to the IgLON5 antibody has a micropolarity. An electromagnetic excitation device is provided at the bottom of the detection component, and the electromagnetic excitation device is utilized to cause at least a portion of the detection component to generate an electromagnetic effect, thereby affecting the effect of the target marker adhering to the surface of the detection component.

[0013] Preferably, the detection component comprises: a substrate and a signal expression layer provided on the substrate; the signal expression layer is located on the surface of the detection component;

[0014] The signal expression layer has a microstructure, including a metal film laid on the substrate, and a plurality of detection bumps arranged on the surface of the metal film: the detection bumps include a bottom end connected to the metal film and a disc-shaped top disk opposite to the bottom end; and at least the detection bumps are processed to adhere to the top disk using IgLON5-specific antigen as a binding agent.

[0015] Preferably, the material of the detection bump is a semiconductor material, a solid-state resistor material or a material with nonlinear resistance characteristics, so that the detection bump has the following characteristics: when the voltage loaded on the detection bump exceeds a voltage threshold, the detection bump is electrically turned on.

[0016] Preferably, the detection element is a photodetector, and further includes other necessary accessories to assist the detection element in capturing photoelectric signals.

[0017] Preferably, utilizing said analysis unit comprises analyzing said image data at a pixel level, comprising identifying one or more light signals in the image data.

[0018] Preferably, the analyzing unit analyzes the image data, including analyzing the position, intensity, spectrum and Raman characteristics of one or more optical signals in the image data.

[0019] Preferably, the analysis unit further includes analyzing parameter changes of one or more optical signals in the image data in a time series, including changes in the intensity, spectrum or Raman characteristics of the optical signal in a time series.

[0020] At the same time, a fluorescent labeling method for detecting IgLON5 antibody IgG is proposed, which is applied to the fluorescent labeling system; the fluorescent labeling method comprises the following steps:

[0021] S100: pretreating the fluorescent marker by chemical modification so that the fluorescent marker carries a weak charge;

[0022] S200: mixing the sample to be tested and the fluorescent marker for reaction, and adding the mixture to the signal expression layer of the detection component, so that the target detection object in the mixture specifically binds to the binding agent on the surface of the signal expression layer;

[0023] S300: fixing the detection assembly to the mounting unit, the detection unit capturing images of light signals generated by the fluorescent substance in multiple regions on the surface of the signal expression layer, and operating and controlling the mounting unit to change the relative position of the detection unit and the detection assembly, thereby detecting light signals in different regions of the signal expression layer;

[0024] S400: Identifying and analyzing the light signal image, including analyzing at least one of the position, intensity, spectrum, and Raman characteristics of the light signal in the image.

[0025] Preferably, in step S300, non-contact electromagnetic excitation is applied to the normal direction of the surface of the detection component, thereby promoting the motion state of the target detection object in the detection component.

[0026] Preferably, based on the real-time monitoring result of the light signal image by the detection unit, the working parameters of the non-contact electromagnetic excitation applied to the detection component are changed; wherein the working parameters of the electromagnetic excitation include the voltage or frequency of the excitation.

[0027] The beneficial effects achieved by the present invention are:

[0028] This technical solution incorporates a signal expression layer within the detection assembly. The nanostructure of the metal film and detection bumps significantly enhances the signal strength of fluorescent markers through localized electromagnetic field enhancement. Furthermore, a non-contact electromagnetic excitation device induces a tiny electric field on the metal film, further optimizing the distribution and binding efficiency of the targeted analyte within the detection region, reducing background noise and improving the detection signal-to-noise ratio and overall system sensitivity.

[0029] The electromagnetic excitation device in this technical solution is configured with two operating modes: a first operating state and a second operating state. By dynamically adjusting the duration of the accelerating electromagnetic field based on the fluorescence or Raman signal captured by the real-time detection unit, the device achieves precise distribution and efficient capture of the targeted analyte within the detection region. The periodic electromagnetic excitation not only accelerates the directional movement of the target marker but also ensures the stable binding of the analyte within the detection region, significantly improving the reliability and efficiency of detection.

[0030] This technical solution uses a detection unit to monitor the intensity, distribution, and time series of fluorescence or Raman signals in real time. This data is fed back to the control module to dynamically adjust the operating parameters of the electromagnetic excitation device. The closed-loop feedback control mechanism in this system ensures the stability of the detection process, reduces signal errors caused by sample differences or environmental changes, and enables the system to maintain high accuracy and stability under different detection conditions. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The present invention can be further understood from the following description in conjunction with the accompanying drawings. The components in the figures are not necessarily drawn to scale, but rather the emphasis is placed on illustrating the principles of the embodiments. In different views, the same reference numerals designate corresponding parts.

[0032] Explanation of serial numbers: 10 - detection component; 100 - signal expression layer; 120 - metal film; 130 - detection bump; 140 - top disk; 160 - binder; 170 - substrate; 300 - mounting unit; 310 - power supply; 320 - induction coil; 400 - detection unit; 500 - computer system; 502 - bus; 504 - processor; 506 - main memory; 508 - read-only memory; 510 - storage device; 512 - display; 514 - input device; 516 - cursor control device; 518 - network device;

[0033] Figure 1 is a schematic diagram of the detection component described in an embodiment of the present invention;

[0034] Figure 2 is a schematic diagram of the installation unit described in an embodiment of the present invention;

[0035] Figure 3 is a schematic diagram of the detection unit described in an embodiment of the present invention;

[0036] Figure 4 Schematic diagram comparing the detection sensitivity of the present invention and the traditional fluorescent labeling system;

[0037] Figure 5 Schematic diagram of a computer system used in the system according to an embodiment of the present invention. DETAILED DESCRIPTION

[0038] In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with its embodiments; it should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. For those skilled in the art, other systems, methods and / or features of the present embodiment will become apparent after reviewing the following detailed description. It is intended that all such additional systems, methods, features and advantages are included in this specification. Included within the scope of the present invention and protected by the appended claims. Additional features of the disclosed embodiments are described in the following detailed description, and these features will be apparent from the following detailed description.

[0039] The same or similar reference numerals in the drawings of the embodiments of the present invention correspond to the same or similar components. In the description of the present invention, it should be understood that if there are terms such as "upper", "lower", "left", "right", etc. indicating an orientation or positional relationship, they are based on the orientation or positional relationship shown in the drawings. This is only for the convenience of describing the present invention and simplifying the description, and does not indicate or imply that the device or component referred to must have a specific orientation. The terms used in the drawings to describe the positional relationship are only for illustrative purposes and cannot be understood as limiting this patent. For those skilled in the art, the specific meanings of the above terms can be understood according to the specific circumstances.

[0040] Example 1: Exemplarily, a fluorescent labeling system for detecting IgLON5 antibody IgG is proposed, the fluorescent labeling system comprising:

[0041] The detection component is configured to construct an environment on the surface of the detection component for specific binding of the IgLON5 antibody and the marker with fluorescent properties;

[0042] a detection unit configured to capture an image of the fluorescent signal generated from the surface of the detection component and convert the image into digital image data;

[0043] a mounting unit, configured to fix the detection assembly and enable the detection assembly and a detection element of the detection unit to move relative to each other, so as to allow the detection element to detect various areas of the detection assembly;

[0044] an analysis unit, configured to process and analyze the image data and output a detection result;

[0045] During pretreatment of the fluorescent marker, the fluorescent marker is chemically modified to impart a weak charge, so that the target marker formed after the fluorescent marker specifically binds to the IgLON5 antibody has a micropolarity. An electromagnetic excitation device is provided at the bottom of the detection component, and the electromagnetic excitation device is utilized to cause at least a portion of the detection component to generate an electromagnetic effect, thereby affecting the effect of the target marker adhering to the surface of the detection component.

[0046] Preferably, the detection component comprises: a substrate and a signal expression layer provided on the substrate; the signal expression layer is located on the surface of the detection component;

[0047] The signal expression layer has a microstructure, including a metal film laid on the substrate, and a plurality of detection bumps arranged on the surface of the metal film: the detection bumps include a bottom end connected to the metal film and a disc-shaped top disk opposite to the bottom end; and at least the detection bumps are processed to adhere to the top disk using IgLON5-specific antigen as a binding agent.

[0048] Preferably, the material of the detection bump is a semiconductor material, a solid-state resistor material or a material with nonlinear resistance characteristics, so that the detection bump has the following characteristics: when the voltage loaded on the detection bump exceeds a voltage threshold, the detection bump is electrically turned on.

[0049] Preferably, the detection element is a photodetector, and further includes other necessary accessories to assist the detection element in capturing photoelectric signals.

[0050] Preferably, utilizing said analysis unit comprises analyzing said image data at a pixel level, comprising identifying one or more light signals in the image data.

[0051] Preferably, the analyzing unit analyzes the image data, including analyzing the position, intensity, spectrum and Raman characteristics of one or more optical signals in the image data.

[0052] Preferably, the analysis unit further includes analyzing parameter changes of one or more optical signals in the image data in a time series, including changes in the intensity, spectrum or Raman characteristics of the optical signal in a time series.

[0053] At the same time, a fluorescent labeling method for detecting IgLON5 antibody IgG is proposed, which is applied to the fluorescent labeling system; the fluorescent labeling method comprises the following steps:

[0054] S100: pretreating the fluorescent marker by chemical modification so that the fluorescent marker carries a weak charge;

[0055] S200: mixing the sample to be tested and the fluorescent marker for reaction, and adding the mixture to the signal expression layer of the detection component, so that the target detection object in the mixture specifically binds to the binding agent on the surface of the signal expression layer;

[0056] S300: fixing the detection assembly to the mounting unit, the detection unit capturing images of light signals generated by the fluorescent substance in multiple regions on the surface of the signal expression layer, and operating and controlling the mounting unit to change the relative position of the detection unit and the detection assembly, thereby detecting light signals in different regions of the signal expression layer;

[0057] S400: Identifying and analyzing the light signal image, including analyzing at least one of the position, intensity, spectrum, and Raman characteristics of the light signal in the image.

[0058] Preferably, in step S300, non-contact electromagnetic excitation is applied to the normal direction of the surface of the detection component, thereby promoting the motion state of the target detection object in the detection component.

[0059] Preferably, based on the real-time monitoring result of the light signal image by the detection unit, the working parameters of the non-contact electromagnetic excitation applied to the detection component are changed; wherein the working parameters of the electromagnetic excitation include the voltage or frequency of the excitation.

[0060] Specifically, as attached Figure 1 As shown, an implementation of the detection component is exemplarily described.

[0061] Preferably, the detection component includes a substrate 170 and a signal expression layer 100 .

[0062] Preferably, the substrate 170 serves as the core support component of the detection assembly, primarily supporting the signal expression layer 100, providing mechanical support and stability, and optimizing signal transmission and amplification. The substrate 170 is required to have a highly flat surface with an RMS roughness of less than 10 nm to ensure uniform placement of components on its surface.

[0063] Preferably, the thickness of the substrate 170 is 0.5 mm or less, and may be 0.3 mm or 0.1 mm, or thinner. The thickness of the substrate 170 needs to have a certain mechanical strength.

[0064] Preferably, the material of the substrate 170 is preferably an insulating material and has a certain thermal conductivity to conduct and dissipate the heat that may be generated by the detection component as quickly as possible. The material can be silicon dioxide, silicon nitride, or aluminum oxide.

[0065] Furthermore, the signal expression layer 100 is used to amplify the electromagnetic signal generated by molecules immobilized on the surface of the signal expression layer 100, including the target marker formed by the specific binding of the fluorescent marker and the IgLON5 antibody. Signal amplification refers only to the increase in the detectability of the signal itself, without significantly increasing the number of detectable molecules.

[0066] After amplification, the signal intensity is at least 100 times greater than that observed on glass, plastic, or flat metal films. The amplified signal is confined to a small area near the surface of the signal expression layer 100, meaning the detection depth is typically 200 nm or less. For fluorescence signals, the typical detection depth is 80 nm or less; for Raman scattering signals, the detection depth is typically 60 nm or less.

[0067] The design of the signal expression layer 100 is based on the principle of electromagnetic field enhancement. By constructing a fine nanostructure on its surface, it significantly amplifies the optical signals generated by target molecules, such as fluorescence or Raman scattering. The signal expression layer 100 comprises a metal film 120, detection bumps 130, and a top disk 140. These structures utilize surface plasmon resonance and local electromagnetic field enhancement to concentrate and amplify the energy of the excitation light.

[0068] Near the microstructure's detection bumps 130 and top disc 140, the local electromagnetic field intensity is significantly enhanced, forming a hotspot. This hotspot, located within a detection depth of approximately 150 nm from the surface of the metal film 120, effectively enhances the intensity of the target molecule signal while reducing background noise interference. Furthermore, the surface of the signal expression layer 100, preferably near the top disc 140, is immobilized with an IgLON5-specific antigen as a binding agent via a molecular linker layer, ensuring that the target marker specifically binds and generates a signal within the hotspot, further enhancing detection sensitivity.

[0069] The Signal Expression Layer 100 offers multiple advantages. Its high sensitivity significantly enhances the signal intensity of target molecules compared to traditional detection methods. Furthermore, its localized enhancement significantly reduces background noise, improving the signal-to-noise ratio. Signal amplification by the Signal Expression Layer 100 is linearly proportional to target molecule concentration, making it suitable for concentration detection over a wide dynamic range. It is also compatible with multiple detection methods, including fluorescence, surface-enhanced Raman scattering (SERS), and electroluminescence (ECL), providing strong technical support for high-precision molecular detection.

[0070] More specifically, in an exemplary embodiment, the signal expression layer 100 includes a surface covered with a metal film 120. The metal film 120 is a continuous metal conductive layer; the metal film 120 is laid on the surface of the substrate 170 and connected to the bottom of the detection bump 130. The metal film 120 is preferably made of gold, silver, or copper. In a more preferred embodiment, an alloy metal with excellent conductivity and oxidation resistance, such as a gold-aluminum alloy or a copper-nickel alloy, is used. The metal film 120 serves as the induction device required for subsequent electromagnetic excitation and can also enhance the electromagnetic field strength near the detection bump to amplify the fluorescence or Raman signal.

[0071] In an exemplary embodiment, a plurality of detection bumps 130 are uniformly or non-uniformly arranged on the surface of the metal film 120. The arrangement of the detection bumps 130 can be periodic or non-periodic. Preferably, the height of the detection bumps 130 is 5 nm to 300 nm, more preferably 50 nm to 200 nm. Preferably, the spacing between the detection bumps 130 is less than the wavelength of the light used for the laser fluorescent marker, and preferably generally less than 1 / 2 of the target light wavelength, to ensure that the local electromagnetic field can effectively couple and enhance the signal. Optionally, if the fluorescent marker is fluorescein isothiocyanate, its excitation wavelength is approximately 488 nm; if the fluorescent marker is Cy5, its excitation wavelength is approximately 650 nm. Therefore, when using visible light excitation, the spacing between the detection bumps 130 is 200 nm to 400 nm; when using near-infrared light excitation, the spacing between the detection bumps 130 is 300 nm to 500 nm.

[0072] The detection bump 130 can elevate the signal generation area to a certain height from the metal film, thereby optimizing the distribution of the electromagnetic field.

[0073] Exemplarily, the top disk 140 can be circular, conical, polygonal, elliptical, or elongated. The shapes of multiple top disks 140 can be the same or different. The diameter of the top disk 140 is preferably in the range of 50 nm to 200 nm. Within the visible light range (400 nm to 700 nm), the recommended diameter is 80 nm to 120 nm, and within the near-infrared range (700 nm to 1000 nm), the recommended diameter is 120 nm to 200 nm. The surface plasmon resonance effect is optimal when the diameter of the top disk 140 is close to 1 / 4 to 1 / 2 the wavelength of the excitation light. This design effectively concentrates the electromagnetic field energy, forming a high-intensity localized hotspot in the gap between the top disk 140 and the metal film 120, significantly enhancing the optical signal generated by the fluorescent marker. A smaller diameter top disk 140 is suitable for densely distributed nanostructures, but this will relatively limit the overall signal amplification effect. A larger diameter top disk 140, on the other hand, can cover a wider electromagnetic field area, but the hotspot density will be reduced. Therefore, the diameter design of the top disk 140 needs to strike a balance between hotspot density and electromagnetic field enhancement effect.

[0074] Preferably, the thickness of the top disk 140 affects the signal amplification performance and structural stability, and its preferred range is 5nm to 80nm. In visible light detection, the recommended thickness of the top disk 140 is 10nm to 30nm, and in near-infrared light detection, the recommended thickness is 30nm to 50nm. A thinner top disk 140 helps to produce a stronger surface plasmon resonance effect, but may cause thermal effects or structural instability under high-intensity excitation. Although a thicker top disk 140 slightly reduces the resonance effect, it can significantly improve the mechanical stability of the structure and maintain a higher resonance effect under specific circumstances.

[0075] Preferably, the material of the top disk 140 can be the same as that of the metal film 120 .

[0076] Furthermore, a binding agent 160 is coated on at least a portion of the surface of the top disk 140 and the detection bump 130. Preferably, the binding agent 160 is a reagent that can specifically bind to the IgLON5 antibody by interacting with the IgLON5 antibody. The specific binding enables the binding agent 160 to bind to and concentrate the IgLON5 antibody from a test mixture containing various molecules; the specific binding effect is usually mediated by the affinity region of the binding agent. Preferably, the binding agent 160 can be any molecule that can specifically bind to the IgLON5 antibody. Preferably, the binding agent 160 can be selected to have a dissociation constant of less than 10-6 M and does not significantly bind other molecules.

[0077] Specific binding refers to the ability of a binding agent to preferentially bind to IgLON5 antibodies in a heterogeneous mixture. Specific binding interactions are able to distinguish the desired target molecule in the sample, i.e., IgLON5 antibodies or IgLON5 aptamers, with a binding difference of 50-fold or greater between the target molecule and non-target molecules, resulting in a clearly resolvable detection effect.

[0078] Exemplarily, the binding agent 160 can be a full-length or partial fragment of IgLON5 protein, such as a genetically engineered recombinant IgLON5 protein or a specific antigenic fragment of IgLON5 protein, or a synthetic IgLON5 peptide.

[0079] Exemplarily, the binder 160 can be attached to the surface of the signal expression layer by one or more of the following methods: (1) through a molecular linker layer, such as using a molecule with amino NH2 or sulfhydryl SH functionality to connect the binder to the surface of the signal expression layer; (2) through chemical modification, such as connecting the capture agent to the signal expression layer through NHS esters, affinity molecules or oligonucleotide probes.

[0080] Furthermore, the mounting unit 300 is used to fix the detection component and provide a high-precision relative movement function to ensure that the detection unit can cover various areas of the detection component for signal acquisition. Preferably, the mounting unit 300 includes a stable mechanical structure that can keep the position of the detection component precisely controllable during the detection process, avoiding deviation or distortion of the detection results due to slight vibrations or displacements. Preferably, the mounting unit 300 includes an adjustable mechanical platform that can perform high-precision fine-tuning in three orthogonal directions of x, y, and z to achieve precise alignment and area coverage between the detection component and the detection unit.

[0081] The mounting unit 300 drives the detection assembly through a precision transmission mechanism, such as a servo motor, magnetic guide rail, or motorized platform, to perform relative displacement. This displacement can be continuous or point-by-point, depending on the required detection accuracy and pixel resolution. The transmission mechanism is controlled by a control module within the mounting unit. This control module adjusts the detection assembly's movement speed, direction, and step size based on real-time feedback from the detection unit, ensuring that the detection unit consistently covers each detection area of ​​the detection assembly.

[0082] Preferably, the mounting unit 300 is also equipped with a positioning and feedback system, such as an optical sensor or encoder, for real-time monitoring of the detection component's position information, ensuring that the detection component accurately stays in the predetermined position after each movement, thereby improving the accuracy and stability of the detection. During the detection process, the mounting unit 300 ensures optical alignment between the detection component and the detection unit, preventing inaccurate optical signal readings due to positional deviations.

[0083] Preferably, the mechanical structure of the mounting unit 300 also needs to have good vibration resistance and thermal stability to avoid slight mechanical deformation or thermal expansion during long-term testing or in complex environments, thereby affecting the detection accuracy.

[0084] For example, as shown in the attached Figure 2 As shown, an electromagnetic excitation device is provided at an appropriate distance from the bottom of the detection assembly; preferably, the electromagnetic excitation device includes an induction coil 320, a power supply 310, and a control module for controlling the operation of the electromagnetic excitation device. The induction coil 320 is used to generate a small current or voltage in the metal film 120 through electromagnetic induction without directly contacting the detection assembly. The electromagnetic excitation device generates an alternating electromagnetic field around itself, causing an induced current to be generated on the surface of the detection assembly, particularly the metal film 120, thereby forming a local electromagnetic field enhancement effect at least near the surface of the metal film 120. The electromagnetic effect can effectively influence the target markers attached to the surface of the detection assembly, optimizing their attachment and positioning.

[0085] Preferably, the power supply 310 is an AC power supply or a DC power supply equipped with an AC output conversion function. The power supply 310 provides the induction coil 320 with alternating electrical energy to generate an induced current, so that the induction coil 320 can continuously generate a stable alternating electromagnetic field. The output parameters of the power supply 310, including voltage, frequency, and current intensity, can be adjusted according to actual needs to meet the electromagnetic excitation requirements in different detection scenarios. The power supply 310 typically includes an adjustable output module for precisely controlling the voltage and frequency applied to the induction coil 320 to ensure the stability and controllability of the electromagnetic field.

[0086] Specifically, the electromagnetic excitation device adjusts the distribution and intensity of the electromagnetic field by controlling the frequency and intensity of the current flowing through the induction coil 320, thereby controlling the current or voltage generated by the metal film 120. While the tiny current or voltage generated does not directly affect the physical state of the detection component, it can stimulate local electric and electromagnetic fields on the surface of the detection component, thereby enhancing the interaction between the target marker and the metal film. Appropriate electromagnetic excitation can promote the directional attachment of the target marker, strengthen its binding to the detection bump 130, and optimize the signal enhancement effect.

[0087] Under the action of the electromagnetic excitation device, the tiny electric field generated on the surface of the metal film can significantly affect the movement of the charged IgLON5 antibody IgG as the target analyte before and after specific binding. The effect of the electric field on the target analyte is similar to the electrophoresis effect, where charged molecules will move in a direction under the action of the electric field. When the electric field intensity is high, the target analyte will be guided to accumulate on the surface of the metal film, especially in areas with strong electric fields, thereby increasing its probability of binding to the capture agent on the surface of the metal film. The movement speed of the target analyte is positively correlated with the intensity of the electric field. The stronger the electric field, the more significant the aggregation of the target analyte. This process effectively improves the attachment efficiency of the target marker, especially in low-concentration samples, and can enhance the sensitivity and stability of detection.

[0088] In addition to its directional effect, the electric field also enhances the binding force between the target analyte and the surface of the metal film 120 by increasing the distribution of localized charge on the surface of the metal film 120. When the target analyte (such as IgLON5 antibody IgG) reaches the metal film surface under the guidance of the electric field, the surface electric field enhances electrostatic adsorption, promoting the binding of the target marker to the metal film. The electric field strengthens the surface electrostatic force or dipole interaction, further enhancing the adhesion of the target analyte, thereby forming a more stable bond to the surface. In high-concentration samples, the electric field prevents the target marker from falling off the metal film surface, reducing signal loss and improving signal stability and detectability.

[0089] In addition, the tiny electric field generated by the metal film 120 not only facilitates the directional attachment of the target marker, but also further enhances the intensity of the local electromagnetic field by affecting the electromagnetic field near the surface of the metal film 120. This enhanced electric field effect significantly increases the intensity of the optical signal generated by the target analyte, such as fluorescence or Raman signal, through the surface plasmon resonance effect. In fluorescence detection, the enhanced electric field causes the target marker to emit a more obvious light signal, thereby improving detection sensitivity and accuracy; while in Raman scattering detection, the enhanced effect of the electric field increases the scattering ability of the molecules to light, further increasing the intensity of the Raman signal and obtaining clearer detection results. These effects not only enhance the signal intensity of the detection, but also increase the accuracy of the quantitative analysis, reduce errors caused by signal instability or marker desorption, and ensure efficient and stable quantitative analysis.

[0090] And in a preferred embodiment, the electromagnetic excitation device is also fixed to the installation unit 300 and driven by the installation unit 300 to move synchronously with the detection component.

[0091] For example, as shown in the attached Figure 3As shown, the detection unit 400 is disposed above the detection assembly and is used to capture and analyze optical signals generated from the surface of the detection assembly. Preferably, the detection unit 400 includes a photodetector, optionally a photodiode, a photomultiplier tube, or a CCD camera, for capturing fluorescence or Raman signals generated on the surface of the detection assembly. These photodetectors can convert optical signals into electrical signals for subsequent processing by the analysis unit.

[0092] To further optimize signal capture and reduce background noise, detection unit 400 also includes auxiliary optical components such as optical filters, lenses, and condensers. Optical filters selectively transmit light signals within a specific wavelength range while rejecting irrelevant wavelengths, thereby improving signal purity. Lenses and condensers adjust the focus and transmission of the light beam, ensuring that the light signal is accurately projected onto the detector for optimal light signal collection.

[0093] Embodiment 2: This embodiment should be understood to include at least all the features of any of the above embodiments, and further improve upon them:

[0094] In an exemplary embodiment, the electromagnetic excitation device utilizes a periodic electromagnetic excitation control algorithm to achieve periodic acceleration of the targeted analyte, and is combined with a detection unit to perform real-time signal detection. The electromagnetic excitation control algorithm has two operating states, alternatingly adjusting the excitation intensity of the electromagnetic excitation device within a specific period to achieve periodic acceleration of the targeted analyte.

[0095] Preferably, the two working states include a first working state and a second working state, and in addition to the two working states, the electromagnetic excitation device can also be set to stop working, that is, not generate any electromagnetic excitation effect. The duration of the first working state is set to t1, and the duration of the second working state is set to t2.

[0096] In some embodiments, t1>t2, so that the targeted analyte is under the action of a stable basic electromagnetic field; and t2 has a shorter duration, providing a brief acceleration effect of the accelerating electromagnetic field.

[0097] Preferably, in the first working state, the electromagnetic excitation device generates minimal excitation, maintaining a basic electromagnetic field for attracting and directing the target analyte. At this time, the target analyte slowly moves toward the surface of the metal film under the action of the micro electric field.

[0098] Preferably, during time t2, the electromagnetic excitation device enters a second operating state, in which it generates a stronger electromagnetic field that acts on the targeted analyte, accelerating its movement. This state lasts for a short time, intended to provide a brief acceleration of the targeted analyte, helping it quickly reach the detection area or increasing its frequency of contact with the capture agent.

[0099] For example, the voltage range of the electromagnetic excitation device applied to the induction coil 320 should be sufficient to generate an electric field strength sufficient to affect the metal film and stimulate the accelerated movement of the targeted analyte. In a preferred embodiment, in the first operating state, the voltage range is 0.1V to 50V; in the second operating state, the voltage range is 5V to 50V.

[0100] Preferably, the action process time of the second working state is very short, and the holding time t2 is usually tens of milliseconds to hundreds of milliseconds.

[0101] Furthermore, the frequency of the electromagnetic excitation device is preferably set to 100kHz to 10MHz. In the first working state, the frequency setting range can be 100kHz to 1MHz, so as to be suitable for the basic electromagnetic field effect at a lower electric field strength, and the electric field can more gently affect the targeted analyte.

[0102] In the second working state, the frequency setting range is 1 MHz to 10 MHz, which is used to generate a stronger accelerating electromagnetic field. The high-frequency electric field can make the analyte respond more quickly, thereby achieving an acceleration effect in a short time.

[0103] In some embodiments, at least one pre-detection is performed, and the value of t2 in the second working state is calculated based on the result of the pre-detection.

[0104] In this pre-test, a sample of known concentration and a fluorescent marker are used, and the specific binding process between them is carried out in the first operating state. This pre-test provides the system with the following key information after specific binding of the sample under the current detection components and operating parameters: the distribution of the target analyte on the metal film surface; the baseline intensity of the fluorescence or Raman signal; and the binding efficiency between the analyte and the capture agent, which can be indirectly inferred from indicators such as fluorescence intensity and signal distribution. This information provides a basis for adjusting the operating state in subsequent tests, ensuring the accuracy and sensitivity of the detection process.

[0105] Furthermore, in the subsequent second detection, the holding time t2 of the second working state of the electromagnetic excitation device is dynamically adjusted according to the results of the pre-detection. Since the concentration of the relevant analyte is known in the pre-detection, the binding efficiency E of the target analyte can be quantitatively determined. binding And calculate the value of t2 using the following formula:

[0106] ;

[0107] In the above formula, k is the adjustment coefficient, which can be optimized after multiple experiments; Va is the migration speed of the target analyte, ΔS is the change in signal intensity per unit time (such as the change in fluorescence intensity), and E binding is the target analyte binding efficiency measured during pre-test.

[0108] Preferably, when the pre-detected binding efficiency E binding If the electromagnetic field is low, the system will appropriately increase the duration of t2 to accelerate the effect of the electromagnetic field on the targeted analyte, prompting it to locate on the surface of the signal expression layer 100 more quickly.

[0109] If the signal intensity change ΔS of the first detection is low, it means that the binding or orientation process of the target analyte is not ideal. At this time, the time t2 of the accelerating electromagnetic field can be increased to accelerate the attachment and orientation of the target marker.

[0110] If the signal strength and binding efficiency are good, t2 in the second detection can be shortened appropriately to maintain a stable electromagnetic field effect.

[0111] In a preferred embodiment, detection bump 130 is constructed of a semiconductor material whose conductivity is affected by the applied voltage, resulting in detection bump 130 exhibiting distinct switching characteristics under varying voltage conditions. Specifically, when the voltage does not reach a specific threshold, detection bump 130 is in a non-conductive state. However, when the applied voltage exceeds the threshold, detection bump 130 becomes electrically conductive. In this non-conductive state, top disk 140 is non-inductive, and the electric field effect between it and metal film 120 is weak. The electromagnetic field enhancement effect in the local hotspot region is limited, and the aggregation and binding of targeted analytes primarily relies on the electric field attraction on the surface of metal film 120.

[0112] When the detection requirements require higher target analyte capture efficiency or signal enhancement effect, the induced voltage of the metal film 120 can be increased so that it reaches or exceeds the conduction threshold voltage of the detection bump 130. At this time, the detection bump 130 will enter the on state, and the current can flow between the metal film and the detection bump, and form a significant electric field enhancement area at the metal disk on the top of the detection bump. This electric field effect not only significantly increases the aggregation rate of the target analyte at the top of the detection bump, but also can improve the binding efficiency and signal strength of the target marker in a shorter time. Especially in the detection scenario of low-concentration target analytes, this local electric field enhancement can effectively overcome the limitations of molecular diffusion and improve the success rate of molecular capture.

[0113] The material selection for the detection bump 130 needs to be appropriately matched to the induced voltage setting of the metal film to ensure precise control within a specific voltage range. Preferably, the detection bump 130 is made of a semiconductor material, such as silicon, gallium arsenide, or gallium nitride. These materials have excellent switching characteristics and stable electric field response, exhibiting distinct conducting and non-conducting states under different voltage conditions. The doping concentration, bandgap width, and breakdown voltage of these materials need to be appropriately designed to ensure that their conduction threshold is within a controllable range (e.g., 1V to 10V). This prevents premature conduction under normal operating conditions while ensuring effective conduction when required.

[0114] In addition, the induced voltage setting of the metal film 120 must be optimized through accurate electromagnetic field simulation and calculation. The amplitude and distribution of the induced voltage must meet the following requirements:

[0115] Ensure that when the voltage does not reach the conduction threshold, the detection bump remains in a non-conducting state to maintain the basic electromagnetic field attraction effect.

[0116] When the voltage exceeds the conduction threshold, the metal disk on top of the detection bump can generate an effective electric field enhancement, significantly improving the aggregation and binding efficiency of the target marker.

[0117] The tagging system dynamically adjusts the induced voltage setting based on real-time feedback from the detection unit, ensuring optimal matching of the detection bump's conduction state and electric field strength under varying detection conditions. This dynamic coupling of voltage and material properties allows for flexible adjustment based on diverse detection requirements, preventing material breakdown due to excessive voltage while ensuring efficient electromagnetic excitation for targeted analyte capture and signal amplification.

[0118] As attached Figure 4 The following graph shows a comparison of the detection sensitivity of this technology solution and traditional fluorescent labeling systems. The horizontal axis represents analyte concentration, using a logarithmic scale, representing target analyte concentrations ranging from 1 fM to 100 nM. The vertical axis represents fluorescence signal intensity (au), indicating the detected fluorescence signal intensity. Lower values ​​indicate lower concentrations that can be detected at the same detection intensity, indicating greater detection sensitivity.

[0119] By comparison, at low concentrations (fM to pM), the traditional detection scheme has weak signal intensity and the curve is close to horizontal; at medium and high concentrations (pM to nM), the signal intensity gradually increases, but the enhancement is limited.

[0120] This technical solution (solid line) exhibits significantly higher fluorescence signal intensity at low concentrations (fM to pM), with the curve showing a significant rise at low concentrations. In the medium-to-high concentration range (pM to nM), the fluorescence signal intensity gradually approaches saturation, but the overall level remains higher than that of traditional solutions.

[0121] Embodiment 3: This embodiment should be understood to include at least all the features of any of the above embodiments, and further improve upon them:

[0122] For example, the Figure 5 A schematic diagram of a computer system 500 in which the fluorescent labeling system described herein can be implemented is depicted; the computer system 500 can control the operation of each working unit, module, and component in the system according to the current control program; and also includes the collection, storage, and processing of working data and detection data generated during the operation of the system to ultimately achieve the expected effect of the fluorescent labeling system.

[0123] The computer system 500 includes a bus 502 or other communication mechanism for transmitting information, and one or more processors 504 coupled to the bus 502 for processing information; the processor 504 may be, for example, one or more general-purpose microprocessors;

[0124] The computer system 500 also includes a main memory 506, such as a random access memory (RAM), a cache, and / or other dynamic storage device, coupled to the bus 502 for storing information and instructions to be executed by the processor 504; the main memory 506 may also be used to store temporary variables or other intermediate information during execution of instructions to be executed by the processor 504; these instructions, when stored in a storage medium accessible to the processor 504, present the computer system 500 as a special-purpose machine customized to perform the operations specified in the instructions;

[0125] The computer system 500 may also include a read-only memory (ROM) 508 or other static storage device coupled to the bus 502 for storing static information and instructions for the processor 504; a storage device 510 such as a magnetic disk, an optical disk, or a USB drive (flash drive) is coupled to the bus 502 for storing information and instructions;

[0126] And further, coupled to the bus 502 may also include a display 512 for displaying various information, data, media, etc., an input device 514 for allowing a user of the computer system 500 to control, manipulate, and / or interact with the computer system 500;

[0127] A preferred way of interacting with the management system may be through a cursor control device 516, such as a computer mouse or similar control / navigation mechanism;

[0128] Furthermore, the computer system 500 may further include a network device 518 coupled to the bus 502; wherein the network device 518 may include, for example, a wired network card, a wireless network card, a switching chip, a router, a switch, and other components;

[0129] In general, the terms "engine," "component," "system," "database," and the like as used herein may refer to logic embodied in hardware or firmware, or to a collection of software instructions, possibly with entry and exit points, written in a programming language such as Java, C, or C++; software components may be compiled and linked into executable programs, installed in a dynamic link library, or may be written in an interpreted programming language (e.g., BASIC, Perl, or Python); it will be understood that software components may be callable from other components or from themselves, and / or may be called in response to detected events or interrupts;

[0130] Software components configured to execute on a computing device may be provided on a computer-readable medium, such as a compact disc, digital video disc, flash drive, magnetic disk, or any other tangible medium, or as a digital download (and may be initially stored in a compressed or installable format that requires installation, decompression, or decryption prior to execution); such software code may be stored in part or in whole on a memory device of the executing computing device for execution by the computing device; software instructions may be embedded in firmware, such as an EPROM. It should also be understood that hardware components may be composed of connected logic units (such as gates and flip-flops), and / or may be composed of programmable units (such as programmable gate arrays or processors);

[0131] Computer system 500 includes a processor that can implement the techniques described herein using custom hard-wired logic, one or more ASICs or FPGAs, firmware, and / or program logic that, in combination with the computer system, renders computer system 500 a special-purpose computing device;

[0132] According to one or more embodiments, the techniques herein are performed by computer system 500 in response to processor 504 executing one or more sequences of one or more instructions contained in main memory 506; such instructions may be read into main memory 506 from another storage medium, such as storage device 510; execution of the sequences of instructions contained in main memory 506 causes processor 504 to perform the process steps described herein; in alternative embodiments, hard-wired circuitry may be used in place of or in combination with software instructions;

[0133] As used herein, the term "non-transitory media" and similar terms refer to any media that store data and / or instructions that cause a machine to operate in a specific fashion; such non-transitory media may include non-volatile media and / or volatile media; non-volatile media include, for example, optical or magnetic disks, such as storage device 510; volatile media include dynamic memory, such as main memory 506;

[0134] Among these, common forms of non-transitory media include, for example, floppy disks, diskettes, hard disks, solid-state drives, magnetic tape or any other magnetic data storage medium, CD-ROMs, any other optical data storage medium, any physical medium having a pattern of holes, RAM, PROM and EPROM, FLASH-EPROM, NVRAM, any other memory chip or cartridge, and networked versions thereof;

[0135] Non-transient media are distinct from, but may be used in conjunction with, transmission media; transmission media participate in the transmission of information between non-transient media; for example, transmission media include coaxial cables, copper wires, and optical fibers, including the wires that comprise bus 502; transmission media may also take the form of sound or light waves, such as radio waves and infrared data communications.

[0136] Although the present application has been described above with reference to various embodiments, it will be understood that many changes and modifications may be made without departing from the scope of the present application. That is, the methods, systems, and devices discussed above are examples. Various configurations may omit, replace, or add various processes or components as appropriate. For example, in alternative configurations, the methods may be performed in an order different from that described, and / or various components may be added, omitted, and / or combined. Moreover, features described with respect to certain configurations may be combined in various other configurations, such as different aspects and elements of the configurations may be combined in a similar manner. Furthermore, as technology develops, the elements therein may be updated, i.e., many of the elements are examples and do not limit the scope of the present disclosure or the claims.

[0137] Specific details are given in the description to provide a thorough understanding of the exemplary configurations, including implementations. However, the configurations can be practiced without these specific details. For example, well-known circuits, processes, algorithms, structures, and techniques have been shown without unnecessary detail to avoid obscuring the configurations. This description provides only example configurations and does not limit the scope, applicability, or configurations of the claims. Instead, the foregoing description of the configurations will provide those skilled in the art with an enabling description for implementing the described techniques. Various changes may be made to the function and arrangement of the elements without departing from the spirit or scope of the present disclosure.

[0138] In summary, it is intended that the above detailed description be considered illustrative rather than restrictive, and it should be understood that the above embodiments are intended to be merely illustrative of the present invention and not to limit the scope of protection of the present invention. After reading the contents of the present invention, a skilled person may make various changes or modifications to the present invention, and these equivalent changes and modifications also fall within the scope defined by the claims of the present invention.

Claims

1. A fluorescent labeling system for detecting IgLON5 antibody IgG, characterized in that: The fluorescent labeling system comprises: The detection component is configured to construct an environment on the surface of the detection component for specific binding of the IgLON5 antibody and the fluorescent marker; a detection unit configured to capture an image of the fluorescent signal generated from the surface of the detection component and convert the image into digital image data; a mounting unit, configured to fix the detection assembly and enable the detection assembly and a detection element of the detection unit to move relative to each other, so as to allow the detection element to detect various areas of the detection assembly; an analysis unit, configured to process and analyze the image data and output a detection result; In the pretreatment of the fluorescent marker, the fluorescent marker is chemically modified to have a weak charge, so that the target marker formed after the fluorescent marker specifically binds to the IgLON5 antibody has a slight polarity; an electromagnetic excitation device is provided at an appropriate distance from the bottom of the detection component, and the electromagnetic excitation device is used to cause at least a portion of the detection component to generate an electromagnetic effect, thereby affecting the effect of the target marker adhering to the surface of the detection component; The detection component includes a substrate and a signal expression layer provided on the substrate; the signal expression layer is located on the surface of the detection component; The signal expression layer has a microstructure, including a metal film laid on the substrate, and a plurality of detection bumps provided on the surface of the metal film, wherein the detection bumps include a bottom end connected to the metal film and a top disk in the shape of a disk opposite to the bottom end; and at least the detection bumps are processed to adhere to the top disk using an IgLON5-specific antigen as a binding agent; The electromagnetic excitation device includes an induction coil, a power supply, and a control module for controlling the operation of the electromagnetic excitation device; The electromagnetic excitation device uses a periodic electromagnetic excitation control algorithm to achieve periodic acceleration of the targeted analyte and performs real-time signal detection in conjunction with a detection unit. The electromagnetic excitation control algorithm is configured with two working states, which alternately adjust the excitation intensity of the electromagnetic excitation device within a specific period. The two working states include a first working state and a second working state, and in addition to the two working states, the electromagnetic excitation device is also configured to stop working, i.e., not generate any electromagnetic excitation effect. The duration of the first working state is set to t1, and the duration of the second working state is set to t2, where t1>t2. In the first working state, the electromagnetic excitation device generates minimal excitation, maintaining a stable basic electromagnetic field to attract and orient the targeted analyte. During time t2, the electromagnetic excitation device enters the second working state, at which time the electromagnetic excitation device generates a stronger electromagnetic field to act on the targeted analyte, accelerating its movement and helping the targeted analyte to quickly reach the detection area or increase its contact frequency with the binding agent. Perform at least one pre-detection, and calculate the value of t2 in the second working state based on the result of the pre-detection; In the pre-test, the concentration of the analyte is known, so the binding efficiency E of the target analyte can be quantitatively determined. binding Calculate the value of t2 using the following formula: ； In the above formula, k is the adjustment coefficient, which is optimized after multiple experiments; Va is the migration speed of the target analyte, ΔS is the change in signal intensity per unit time, and E binding is the target analyte binding efficiency measured during pre-test; The material of the detection bump is semiconductor material.

2. The fluorescent labeling system according to claim 1, wherein: The analysis unit also includes analyzing parameter changes of one or more optical signals in the image data in a time series, including changes in the intensity, spectrum or Raman characteristics of the optical signal in a time series.

3. The fluorescent labeling system according to claim 2, wherein: The detection element is a photodetector.

4. The fluorescent labeling system according to claim 3, wherein: The analysis unit includes analyzing the image data at a pixel level, including identifying one or more light signals in the image data.

5. The fluorescent labeling system according to claim 4, wherein: The analyzing unit analyzes the image data, including analyzing the position, intensity, spectrum and Raman characteristics of one or more optical signals in the image data.

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