Optical detection method, device and system

Abstract

The invention discloses an optical detection method, device and system, and relates to the technical field of optical detection. The method comprises the following steps: emitting at least two excitation beams with different wavelengths to a to-be-detected sample, exciting the to-be-detected sample to generate a to-be-detected optical signal, the to-be-detected sample comprising a plurality of luminescent materials with homogeneous systems; receiving a to-be-detected optical signal from the to-be-detected sample in a plurality of preset gating time windows; converting the optical signal to be detected into an electric signal; acquiring light intensity information of the to-be-detected optical signal in each gating time window according to the electric signal; calculating the initial light-emitting signal intensity of the light-emitting material of each homogeneous system according to the light intensity information; and calculating the concentration of the target object in the to-be-detected sample according to the initial light-emitting signal intensity. According to the optical detection method provided by the embodiment of the invention, multi-wavelength detection of the to-be-detected sample can be realized, the concentration of each target object in the to-be-detected sample is obtained, and the detection efficiency is greatly improved.

Description

Technical Field

[0001] The present invention relates to the field of optical detection technology, and in particular to an optical detection method, equipment and system. Background Art

[0002] In many fields such as biomedical research, environmental monitoring and materials science, as the demand for rapid and accurate detection of multiple targets simultaneously grows, existing detection technologies are facing increasing challenges. Currently, the single excitation and receiving path design commonly used in homogeneous detection equipment can no longer meet the requirements of high-efficiency and high-throughput detection.

[0003] First, conventional fluorescence spectrometers can usually only detect light of specific wavelengths sequentially, which requires manual or mechanical replacement of filters when detecting different wavelengths. This operation is not only time-consuming, but also increases the complexity of the experiment and limits the throughput and continuity of the detection. In addition, due to the limitations of the physical size and optical properties of the filter, this method is difficult to achieve high spectral resolution detection, which is particularly prominent when it is necessary to accurately distinguish very close spectral features. Secondly, although time-resolved fluorescence analysis can provide information on the luminescence lifetime of the sample, its implementation usually relies on complex time-resolved systems such as fast gating or single-photon counting technology. These systems are not only costly, but also complex to operate and require professional technicians for maintenance and calibration. In addition, the time resolution of these systems is usually limited by the response speed of electronic components, which is difficult to meet the research needs of ultra-fast processes. Furthermore, the existing technology has challenges in the synchronous detection of multi-color light signals. Since the excitation and emission characteristics of light of different wavelengths in the sample may be different, precise optical path design and time control are required to ensure that all wavelengths of light signals can be detected synchronously and accurately. This is technically very challenging, especially when high time resolution and high spectral resolution are required. Finally, another limitation of existing technologies is their flexibility and adaptability. With the continuous discovery of new biomarkers and environmental pollutants, and the continuous development of new materials, researchers need systems that can quickly adapt to new detection targets. However, existing detection systems often lack the necessary flexibility and are difficult to adapt to new detection needs, which limits their application in emerging fields.

[0004] In summary, the prior art has significant technical problems in the rapid and accurate detection of multiple targets, which are difficult to solve due to the physical limitations of optical components, the response speed of electronic systems, and the complexity of system design. Summary of the invention

[0005] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention proposes an optical detection method, device and system, which can realize multi-channel simultaneous detection and improve detection efficiency.

[0006] In a first aspect, an optical detection method according to an embodiment of the present invention comprises the following steps:

[0007] Emitting at least two excitation light beams of different wavelengths to a sample to be tested, to excite the sample to be tested to generate a light signal to be detected, wherein the sample to be tested includes a plurality of homogeneous system luminescent materials;

[0008] Receiving the optical signal to be detected from the sample to be detected within a plurality of preset gated time windows;

[0009] Converting the optical signal to be detected into an electrical signal;

[0010] Acquiring light intensity information of the optical signal to be detected within each of the gated time windows according to the electrical signal;

[0011] Calculate the initial luminescence signal intensity of the luminescent material of each homogeneous system according to the light intensity information;

[0012] The concentration of the target in the sample to be tested is calculated according to the initial luminescent signal intensity.

[0013] According to some embodiments of the present invention, the time intervals of the plurality of preset gated time windows are set based on the luminescence lifetimes of luminescent materials of a plurality of homogeneous systems.

[0014] According to some embodiments of the present invention, acquiring the light intensity information of the light signal to be detected in each of the gated time windows according to the electrical signal, and calculating the initial luminescent signal intensity of the luminescent material of each homogeneous system according to the light intensity information, comprises:

[0015] Detecting the luminescence intensity decay curves of n homogeneous systems of luminescent materials over time respectively, where n≥2 and n is a positive integer;

[0016] Select n time points t1, t2, ..., t n , respectively calculate the light intensity information of the luminescent material of each homogeneous system at these n time points; and calculate the initial luminescent signal intensity of the target object in the sample to be tested of each homogeneous system based on the light intensity information.

[0017] According to some embodiments of the present invention, the luminescent material is an afterglow material.

[0018] According to some embodiments of the present invention, the structural formula of the luminescent material is as follows:

[0019]

[0020] Wherein, X1 and X2 are selected from one of S or O;

[0021] R1 and R2 are independently selected from at least one of the following groups:

[0022] (a) absent; (b) hydrogen; (c) halogen; (d) amino; or (e) a derivative of amino.

[0023] According to some embodiments of the present invention, the amino derivative is at least one of an N-alkylated amino group, an N,N-dialkylated amino group, an N-arylamino group, and an N,N-diarylamino group.

[0024] According to some embodiments of the present invention, the N-arylamino group is selected from: N-phenylamino group, N-(substituted phenyl)amino group, and N-(condensed ring aryl)amino group.

[0025] According to some embodiments of the present invention, the substituent of the N-(substituted phenyl)amino group is selected from at least one of halogen, alkyl, alkoxy, amino, and nitro.

[0026] According to some embodiments of the present invention, the N-(condensed ring aromatic) amino group is selected from naphthyl, anthracenyl or phenanthrenyl. The optical detection method according to the embodiment of the present invention has at least the following beneficial effects: it can realize multi-wavelength detection of the sample to be tested, obtain the concentration of each luminescent material in the sample to be tested, and greatly improve the detection efficiency. In the second aspect, the optical detection device according to the embodiment of the present invention includes:

[0027] A light source system for emitting at least two excitation light beams of different wavelengths; the excitation light beams of different wavelengths do not interfere with each other during transmission and / or are independent in light beam space;

[0028] A spectroscopic element, used for reflecting and transmitting the excitation light beam, so that the reflected or transmitted excitation light beam is transmitted to the sample to be tested, and excites the sample to be tested to generate a light signal to be detected;

[0029] The detector is used to receive the optical signal to be detected and convert the optical signal to be detected into an electrical signal representing a detection result.

[0030] According to some embodiments of the present invention, the optical detection device also includes a light intensity sensing module, a processing module and a light source controller. The light intensity sensing module is used to detect the output intensity of the excitation light beams of different wavelengths of the light source system, and send the detection results to the processing module; the processing module is used to analyze whether the output intensity of each of the excitation light beams deviates from the preset light source intensity, and send a corresponding adjustment signal to the light source controller based on the excitation light beam with the deviation, so that the light source controller adjusts the output intensity of the corresponding excitation light beam of the light source system.

[0031] According to some embodiments of the present invention, the light source system includes at least two light sources, and the at least two light sources are used to emit excitation light beams of at least two different wavelengths; the optical detection device also includes a fuzzy logic controller and at least two backup light sources, and the backup light sources correspond to the light sources one by one. When the deviation between the output intensity of the excitation light beam of the light source and the preset light source intensity exceeds a threshold, the fuzzy logic controller is used to switch the light source to the corresponding backup light source. The fuzzy logic controller is also used to dynamically adjust the switching speed and response time according to the real-time status of the light source system and system requirements.

[0032] According to some embodiments of the present invention, the optical detection device further includes a collimating element, which is disposed at an output end of the light source system, and is used to convert the excitation light beam into a parallel light beam.

[0033] According to some embodiments of the present invention, the optical detection device also includes a first filter element and / or a second filter element; the excitation light beam is transmitted to the sample to be tested through the first filter element, and the first filter element is used to perform band filtering on the excitation light beam; the second filter element is arranged between the sample to be tested and the detector, and is used to perform band filtering on the light signal to be detected.

[0034] According to some embodiments of the present invention, the optical detection device further comprises a shutter element, wherein the shutter element is disposed between the sample to be detected and the detector, and a switching time of the shutter element is less than 0.2S.

[0035] According to some embodiments of the present invention, the light source system includes at least two light sources, and the at least two light sources are used to emit excitation light beams of at least two different wavelengths; or, the light source system includes a tunable laser, and the tunable laser can emit at least two excitation light beams of different wavelengths under different parameters.

[0036] According to some embodiments of the present invention, the spectroscopic element includes a first spectroscope, a second spectroscope and a third spectroscope; the light source system includes a first light source, a second light source and a third light source, wherein the first excitation light beam emitted by the first light source is transmitted along the horizontal direction, the second excitation light beam emitted by the second light source and the third excitation light beam emitted by the third light source are both perpendicular to the first excitation light beam, and the second light source and the third light source are staggered in the vertical direction, the first spectroscope is arranged at the intersection of the first excitation light beam and the second excitation light beam, the second spectroscope is arranged at the intersection of the first excitation light beam and the third excitation light beam, the first spectroscope and the second spectroscope are used to convert the transmission direction of the second excitation light beam and the third excitation light beam from the vertical direction to the horizontal direction; the third spectroscope is arranged between the light source system and the sample to be tested, and is used to reflect the first excitation light beam and / or the second excitation light beam and / or the third excitation light beam to the sample to be tested; the wavelengths of the first excitation light beam, the second excitation light beam and the third excitation light beam are different.

[0037] According to some embodiments of the present invention, the spectroscopic element also includes a fourth spectroscope and a fifth spectroscope; the detector includes a first detector, a second detector and a third detector, the fourth spectroscope is arranged between the first detector and the sample to be detected, and is used to transmit the light signal to be detected to the first detector, and reflect the light signal to be detected to the fifth spectroscope; the fifth spectroscope is used to reflect the light signal to be detected reflected by the first detector to the second detector, and transmit the light signal to be detected reflected by the first detector to the third detector; the input ends of the first detector, the second detector and the third detector are all provided with filter elements.

[0038] According to some embodiments of the present invention, the optical detection device also includes a focusing lens and / or a collecting lens, wherein the focusing lens is arranged between the spectroscopic element and the sample to be tested, and the sample to be tested is located at the focal position of the focusing lens; the collecting lens is arranged between the sample to be tested and the detector, and is used to collect the light signal to be detected.

[0039] The optical detection device according to the embodiment of the present invention has at least the following beneficial effects: the light source system can emit excitation light beams of different wavelengths simultaneously or sequentially, thereby realizing multi-wavelength detection of the sample to be tested. By partially reflecting and partially transmitting the incident light through the spectroscopic element, the intensity of the optical signal to be detected received by the detector can be effectively improved, thereby enhancing the detection sensitivity and improving the reliability of the signal. In addition, the spectroscopic element can filter out unnecessary optical signals and only allow light of a specific wavelength to pass, thereby reducing interference from optical signals of other wavelengths. The detection device includes multiple detectors, each of which can correspond to a different wavelength, so that multi-channel simultaneous detection can be achieved, greatly improving the detection efficiency.

[0040] In a third aspect, an optical detection system according to an embodiment of the present invention includes:

[0041] The optical detection device as described in the embodiment of the second aspect;

[0042] Testing platform;

[0043] A multi-well plate, disposed on the detection platform, and used for carrying samples to be tested;

[0044] An oscillation module, used for oscillating and mixing the sample to be tested;

[0045] An incubation module, used for incubating the sample to be tested;

[0046] A heating module, used to control the temperature of the incubation module;

[0047] The control module is respectively connected to the optical detection device, the detection platform, the oscillation module, the incubation module and the heating module.

[0048] The optical detection system according to the embodiment of the present invention has at least the following beneficial effects: the porous plate and the aforementioned optical detection device are cleverly integrated together, the optical detection device performs optical detection on the samples to be tested in the porous plate, and the design of the porous plate makes it possible to process multiple groups of samples simultaneously in a smaller space. The incubation module is used to incubate the samples to be tested at a specific temperature, the heating module is used to heat the samples to be tested, and the oscillation module is used to oscillate and mix the samples to be tested. In the entire system, the control module can control the temperature of the heating module, the working mode of the oscillation module, such as the oscillation speed, oscillation time, etc., to meet the needs of different experiments. The design of this optical detection system enables the laboratory to efficiently carry out large-scale sample detection.

[0049] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0051] Figure 1 A schematic diagram of the structure of an optical detection device according to an embodiment of the present invention;

[0052] Figure 2 A schematic structural diagram of an optical detection device according to another embodiment of the present invention;

[0053] Figure 3 is a schematic structural diagram of an optical detection system according to an embodiment of the present invention;

[0054] Figure 4 A schematic diagram of the structure of a porous plate according to an embodiment of the present invention;

[0055] Figure 5 is a flowchart of the steps of the optical detection method according to an embodiment of the present invention;

[0056] Figure 6 Schematic diagram of the detection principle of the optical detection method according to an embodiment of the present invention;

[0057] Figure 7 A schematic diagram of the relationship between the light intensity of a luminescent material and time in an optical detection method according to an embodiment of the present invention;

[0058] Figure 8 is the normalized lifetime decay curve of the luminescent material measured in a dichloromethane solvent at room temperature;

[0059] Fig. 9 is the luminescence intensity decay curve of homogeneous system A and homogeneous system B over time;

[0060] Fig.10 is the luminescence intensity decay curve over time after the homogeneous system A and the homogeneous system B are mixed;

[0061] Fig.11 The standard curves are PCT and IL-6. DETAILED DESCRIPTION

[0062] Embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present application, and are not to be construed as limitations on the present application. For the step numbers in the following embodiments, they are only provided for the convenience of explanation, and the order between the steps is not limited in any way, and the execution order of each step in the embodiment can be adaptively adjusted according to the understanding of those skilled in the art.

[0063] The terms "first", "second", "third" and "fourth" etc. in the specification and claims of the present invention and the drawings are used to distinguish different objects, rather than to describe a specific order. In addition, the terms "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions. For example, a process, method, system, product or device that includes a series of steps or units is not limited to the listed steps or units, but may optionally include steps or units that are not listed, or may optionally include other steps or units that are inherent to these processes, methods, products or devices.

[0064] Reference to "embodiments" in the present invention means that a particular feature, structure, or characteristic described in conjunction with the embodiments may be included in at least one embodiment of the present invention. The appearance of the phrase in various places in the specification does not necessarily refer to the same embodiment, nor is it an independent or alternative embodiment that is mutually exclusive with other embodiments. It is explicitly and implicitly understood by those skilled in the art that the embodiments described herein may be combined with other embodiments.

[0065] The "initial luminescence signal intensity" mentioned in the context of the present invention refers to the luminescence intensity that each luminescent material should theoretically have at the moment of excitation (t=0) obtained by analyzing the light intensity information of the light signal to be detected within multiple preset gated time windows during the optical detection process and after mathematical processing. This intensity value reflects the original luminescence ability of the luminescent material before attenuation and is directly related to the concentration of the target in the sample to be tested.

[0066] The "sample to be tested" mentioned in the context of the present invention refers to a biological or chemical sample to be analyzed and tested, which may contain a target substance to be tested, and is usually a mixture containing multiple components, such as isolated blood, urine, tissue extracts, etc.

[0067] The "target" mentioned in the context of the present invention refers to a specific substance that needs to be specifically detected and quantified in the sample to be tested. The concentration of the target in the sample to be tested is unknown and needs to be determined by the method provided in the embodiment of the present invention.

[0068] The "gated time window" mentioned in the context of the present invention refers to a series of specific time periods set in advance during the optical detection process, which are used to control and limit the acquisition time of the light signal. During these specific time periods, the detection system selectively receives and records the light signals from the sample to be tested, while the signals outside these time periods are ignored or shielded. These time windows are predetermined during the experimental design phase and are set according to the luminescent properties of the substance to be tested and the purpose of the experiment. It usually includes multiple continuous or discontinuous time periods, each of which may have different durations and start times. The setting of multiple gated time windows allows the analysis of the luminescent dynamic characteristics of the luminescent material in different time periods.

[0069] It should be noted that the optical detection equipment, optical detection system and optical detection method of the present application are applied to the field of homogeneous multicolor detection technology. "Homogeneous multicolor" generally refers to the use of markers of different colors (such as fluorescent dyes, quantum dots, etc.) to mark different detection targets in the same reaction system in the field of luminescence, in vitro diagnosis or bioimaging. These markers can be distinguished under specific instruments, thereby achieving simultaneous detection of multiple targets.

[0070] Therefore, homogeneous multicolor detection technology can detect multiple indicators simultaneously in the same reaction system, which greatly improves the detection efficiency and reduces the time of sample processing and experimental operation. Since the detection is completed in the same reaction system, the risk of cross-contamination during sample transfer and operation is reduced. Multicolor labeling technology can also significantly improve the sensitivity and specificity of detection, which helps to identify target molecules more accurately. In homogeneous multicolor detection technology, the design of multi-channel light excitation and multiple detectors is very critical. Multi-channel light excitation can achieve synchronous excitation of multiple markers, thereby realizing the detection of multiple indicators in the same reaction system. Multiple detectors can receive light signals of different wavelengths respectively, realize independent detection of multiple markers, and ensure the accuracy of the detection results. By optimizing the design of multi-channel light excitation and multiple detectors, the detection sensitivity and signal resolution can be improved and the background noise can be reduced.

[0071] However, existing optical systems and detection equipment have technical bottlenecks in the design of multi-path light excitation and multiple detectors, making it difficult to achieve efficient light path allocation and signal reception. In the design of multi-path light excitation and multiple detectors, light signals of different wavelengths may interfere with each other, affecting the accuracy of the detection results.

[0072] To this end, the embodiments of the present invention provide an optical detection device, system and method for a homogeneous multi-color field, which can realize multi-channel excitation and multi-channel reception of light beams, and further realize synchronous detection of multi-channel optical signals. Since the multi-channel optical signals according to the present application can be detected synchronously, multiple groups of samples to be tested can be detected at the same time, thus realizing high-throughput detection of the device and system and shortening the detection time. The optical detection device, system and method according to the present application will be described in detail below in conjunction with the accompanying drawings. It should be understood that the actual optical detection device may also include other components, but in order to avoid blurring the main points of the present application, these other components are not discussed herein and are not shown in the accompanying drawings.

[0073] On the one hand, an embodiment of the present invention proposes an optical detection device, including a light source system, a spectroscopic element and a detector, wherein the light source system is used to emit at least two excitation light beams of different wavelengths, and the excitation light beams of different wavelengths do not interfere with each other during transmission and / or are independent in the light beam space; the spectroscopic element is used to reflect and transmit the excitation light beam, so that the reflected or transmitted excitation light beam is transmitted to the sample to be tested, and excites the sample to be tested to generate a light signal to be detected; the detector is used to receive the light signal to be detected, and convert the light signal to be detected into an electrical signal, thereby sending the electrical signal to a control module, so that the control module analyzes the sample to be tested and obtains a detection result.

[0074] Specifically, in this example, the light source system may include at least two light sources, each of which emits an excitation light beam of a specific wavelength, and the wavelengths of the excitation light beams emitted by different light sources are different. Therefore, multiple light sources may be configured according to the multiple different wavelengths of the required excitation light beams. The light source may be an LED, a laser, etc., which is used to generate an excitation light beam to excite the sample to be tested. The multiple light sources here refer to a combination of light sources of different wavelengths or colors. The combination of multiple light sources can provide multiple fixed wavelengths, and the wavelength of each light source is usually fixed. For example, the first light source may emit an excitation light with a wavelength of 685nm, while the second light source emits an excitation light with a wavelength of 730nm. The selection and configuration of the light source ensure that light of different wavelengths can effectively cover the specific characteristics of the sample to be tested. In addition, in some scenarios, as needed, the multiple light sources here can also be replaced with tunable lasers, that is, the light source system can change the wavelength of the light beam emitted by the tunable laser by adjusting the parameters of the tunable laser. The tunable laser can provide a continuous or specific range of wavelength selection, and the continuous adjustment of wavelength and power can be achieved through a single light source.

[0075] A spectroscopic element is arranged between the light source system and the sample to be tested. The spectroscopic element can be a semi-transparent and semi-reflective mirror, a prism or a diffraction grating. The main function of the spectroscopic element is to receive the incident light from the light source system, and partially reflect and partially transmit it. The reflected / transmitted light can be used for calibration or other auxiliary functions, while the other part of the transmitted / reflected light continues to be transmitted for detecting the sample to be tested. In some embodiments, the spectroscopic element is preferably a semi-transparent and semi-reflective mirror, and the lens of the semi-transparent and semi-reflective mirror preferably has a light transmittance of not less than 98.5% in the wavelength range of 450 to 780 nm.

[0076] It can be understood that in addition to the above-mentioned solution of using a spectroscope, in some application scenarios, the spectroscopic element can also use a multi-channel beam splitter (such as a fiber array or a beam splitter) to distribute the light beam to different lights or channels as needed, or distribute the light beam to different optical fibers through a fiber coupler, and each optical fiber transmits an independent light beam signal, thereby achieving spatial separation; wavelength-selective spectroscopic elements such as diffraction gratings, interference filters or fiber gratings can also be used to separate the different light beams according to their wavelengths, so that light beams of different wavelengths can be transmitted on different paths without interfering with each other.

[0077] In some embodiments, the excitation light beams of different wavelengths emitted by the emission channels of the light source system do not interfere with each other during transmission and / or are independent in the beam space. The "excitation light beams do not interfere with each other during transmission" mentioned here generally refers to the fact that in the same laser emission system, through time division or time-sharing technology, the laser signals of different channels will not interfere with each other during transmission. The "excitation light beams are independent in the beam space" mentioned means that in terms of spatial distribution, the light beams emitted by each laser light source are independent and do not affect each other. It is understandable that this can also be interpreted as multi-wavelength excitation and multi-wavelength emission, which means that the light source used to excite the sample is not a single wavelength, but contains multiple different wavelengths of light. Multi-wavelength emission means that the emission spectrum observed from the sample contains multiple wavelengths. Excitation and emission are orthogonal, which means in optical systems that there is a clear distinction and independence between the excitation wavelength and the emission wavelength. Specifically, this means that the emission spectrum produced by a specific excitation wavelength will not be affected by other excitation wavelengths.

[0078] For example, refer to Figure 1 ,The beam splitting element includes a plurality of beam splitters. By arranging a plurality of beam splitters in the optical path, each beam splitter can split the excitation light beam into two parts, one part continues to be transmitted along the original direction, and the other part is reflected or transmitted to a new direction, for example, Figure 1In the figure, the light beam of the first light source 100 is transmitted along the axially parallel direction. Since the second light source 200 and the third light source 300 are arranged in a direction perpendicular to the light beam of the first light source 100, the light beams of the second light source 200 and the third light source 300 are respectively reflected by the first beam splitter 400 and the second beam splitter 500, thereby changing their original transmission directions from a direction perpendicular to the first excitation light beam emitted by the first light source 100 to a direction parallel to the first excitation light beam. In this way, the samples to be tested in the same area can be excited so that the light signals to be detected are captured by the detector, thereby analyzing the samples to be tested.

[0079] After the sample to be tested receives the excitation light beam, it generates a light signal to be detected. The detector is used to receive and detect the light signal to be detected that passes through the spectroscopic element. The detector can accurately capture the light signal that passes through the spectroscopic element and convert it into an electrical signal for subsequent data processing and analysis, thereby obtaining relevant information of the sample to be tested.

[0080] In some embodiments, the detector is preferably an avalanche photodiode (APD) and / or a photomultiplier tube (PMT). Since APD and PMT can provide high signal-to-noise ratio and low error measurement results, different color components in a multicolor system can be accurately distinguished and quantified. In homogeneous multicolor detection, light of different colors has different intensities, and the high sensitivity of APD and PMT ensures that weaker light signals to be detected can also be effectively detected during the detection process. Generally speaking, avalanche photodiodes and photomultiplier tubes are installed on optical detection equipment, close to or facing the light path between the light source and the sample, so that the detector can receive the light signal to be detected from the sample to be tested. It is understood that in some cases, the front end of the detector is usually equipped with one or more optical coupling elements, such as lenses or gratings, to focus or separate light signals to be detected of different wavelengths. "Optical coupling" here generally refers to the process of transmitting light signals from the light source system to the sample to be tested, and then from the sample to be tested to the detector. In some examples, PMT is usually used for weak light signal detection requiring high sensitivity and low noise, while APD is more suitable for applications requiring high speed, high throughput and a wide dynamic range. For example, when the detected light signal is very weak, such as when performing fluorescence detection, time-resolved fluorescence measurement, or certain types of enzyme-linked immunosorbent assays (ELISA), a color reaction is usually used to detect the binding of antibodies to antigens, and PMT can effectively detect the weak light signals generated by these color reactions. PMT is preferred for its high sensitivity and low noise. PMT can amplify extremely low levels of light signals and is very suitable for detecting low concentrations of target molecules, especially when detecting short-lived fluorescent molecules. In scenarios that require high throughput and rapid detection, such as drug screening, gene expression analysis, etc., APD has a faster response time and a higher linear range. It is understandable that the configuration of the detector here can be either, but there are some differences if the respective advantages of the two must be compared, but in most cases, it does not have much impact on the detection results.

[0081] The above-mentioned optical detection equipment composed of a light source system, a spectroscopic element and a detector, during detection, the light source system emits light in a specific wavelength range as excitation light. The excitation light is irradiated to the sample to be tested, part of the light is absorbed, and the rest of the light is scattered or transmitted. If the sample contains target chemical or biological molecules, fluorescence or color changes may occur after reaction with the reagent. The light processed by the sample enters a spectroscopic element, such as a grating or prism, for wavelength decomposition and separation. The separated light is guided to the detector, and the detector converts the optical signal to be detected into an electrical signal for subsequent analysis. When the detection area contains multiple target samples, the device switches different light sources to emit excitation light of different wavelengths to achieve independent detection of each target. The photon signals (such as fluorescence, afterglow or color changes) generated by chemical reactions or biochemical reactions in the sample to be tested can be accurately captured by the detector and converted into electrical signals.

[0082] In some embodiments, the optical detection equipment also includes a light intensity sensing module, a processing module and a light source controller. The light intensity sensing module is used to detect the output intensity of the excitation light beam of different wavelengths of the light source system, and send the detection result to the processing module; the processing module is used to analyze whether the output intensity of each excitation light beam deviates from the preset light source intensity, and send a corresponding adjustment signal to the light source controller according to the deviated excitation light beam, so that the light source controller adjusts the output intensity of the corresponding excitation light beam of the light source system.

[0083] Assuming that the light source system includes multiple light sources, if the stability of the light source is not high, the accuracy of the measurement result may be affected. For this reason, a light intensity sensing module is integrated in the optical detection equipment. This module can use a highly sensitive photoelectric sensor to accurately capture the slight changes in the intensity of the light source. The light intensity sensing module is used to detect the output intensity of each wavelength light source in real time. For example, the light intensity sensing module is installed at the proximal end of each light source, that is, the initial part of the path of the light source emitting light, but does not affect the path of the light transmitted to the sample. A fixed point is set in the light path, and the light of all light sources must pass through this point. The light intensity sensing module is installed at this point, which can be used to monitor the light intensity passing through this point. The processing module may include an analog-to-digital converter and a processor. The processor may use an MCU, a CPU, a DSP, etc. The light intensity sensing module transmits the real-time monitoring data to the analog-to-digital converter, converts the analog signal of the light intensity into a digital signal; the digital signal is then transmitted to the processor, and the processor analyzes the data and compares it with the preset light source intensity. If the processor detects that the actual light source intensity deviates from the preset light source intensity, it will send an adjustment signal to the corresponding light source controller through the control interface. The light source controller corresponds to the light source one by one. The light source controller adjusts the output of the light source according to the adjustment signal received, such as adjusting the current or changing the driving parameters of the light source to correct the light intensity. Before the optical detection equipment starts detection, each light source is excited in turn to generate an excitation beam. The photoelectric sensor monitors the light intensity of each light source in real time. The light intensity data is transmitted to the processor in real time, and the processor compares the actual light intensity with the preset light intensity. If the intensity of any light source is lower than or higher than the predetermined range, the processor instructs the light source controller to make adjustments, such as increasing or decreasing the current. After adjustment, the light source is detected again until its intensity is adjusted to an appropriate level. This process ensures that the light source intensity is within the optimal range for each detection, thereby ensuring the accuracy and repeatability of the detection results.

[0084] In some embodiments, the optical detection device also includes a fuzzy logic controller and at least two backup light sources, the backup light sources corresponding one-to-one to the light sources in the light source system, and when the deviation between the output intensity of the excitation light beam of the light source and the preset light source intensity exceeds a threshold, the fuzzy logic controller is used to switch the light source to the corresponding backup light source, and the fuzzy logic controller is also used to dynamically adjust the switching speed and response time according to the real-time status of the light source system and system requirements.

[0085] Specifically, if the light intensity sensor module detects that the deviation exceeds the threshold, it means that there may be a problem with the light source. In this case, a redundant light source component is designed. The redundant light source component uses multi-threaded parallel processing technology so that the system can switch to the backup light source in time when a problem with the light source is detected. The fuzzy logic controller is used to dynamically adjust the switching speed and response time according to the real-time status of the light source and system requirements to ensure the continuity and accuracy of the detection.

[0086] For example, each wavelength of the light source in the light source system is equipped with at least one backup light source of the same wavelength. The backup light source is configured in parallel with the main light source, but is in standby mode under normal circumstances. Each pair of light sources (main light source and backup light source) is connected to a light source controller, which is responsible for managing the output and switching of the light sources. Multi-threaded parallel processing technology is used to monitor the status of all light sources at the same time. Each light source (including the backup light source) has an independent monitoring thread, which is responsible for collecting light intensity data and status information in real time. This design ensures the high responsiveness and real-time performance of the system to the light source status, so that it can quickly switch to the backup light source when there is a problem with the main light source. The integrated fuzzy logic controller is used to process the switching logic of the light source. The fuzzy logic controller receives data from the light intensity sensing module and evaluates the health status of the light source according to the preset rules and fuzzy logic algorithm. The controller dynamically adjusts the switching speed and response time to optimize the continuity and accuracy of the detection process, taking into account the real-time status of the light source and the detection requirements of the system. When the controller determines that the main light source needs to be replaced, it sends a switching signal to the light source controller. The light source controller immediately deactivates the main light source and starts the backup light source. The entire switching process can be completed in milliseconds. The switching mechanism also includes a feedback loop where, after the switch is complete, the new light source status is re-evaluated to ensure that it is functioning properly.

[0087] In some embodiments, the optical detection device further includes a collimating element, which is arranged at the output end of the light source system, and the collimating element is used to convert the excitation light beam into a parallel light beam. It is understandable that when the light source system emits an excitation light beam, these light beams may show a certain divergence due to the characteristics of the light source and the design of the optical system. The collimating element is placed at the output end of the light source system, and can focus or adjust the light beam so that the light beam becomes parallel after passing through the collimating element. This parallel light beam can reduce the scattering and loss of the light beam during propagation. By using a collimating element, the number of optical components required can be reduced, thereby simplifying the design and manufacture of the optical system. In some embodiments, the collimating element is preferably a laser lens.

[0088] In some embodiments, the optical detection device further includes a first filter element and / or a second filter element; the excitation light beam is transmitted to the sample to be tested through the first filter element, and the first filter element is used to perform wavelength screening on the excitation light beam; the second filter element is arranged between the sample to be tested and the detector, and is used to perform wavelength screening on the optical signal to be detected. The first filter element and the second filter element are arranged before the excitation light beam emitted by the light source system reaches the detector, and are used to perform wavelength screening on the light beam, so that only light of the required wavelength band is allowed to pass before the light beam reaches the detector. Among them, the first filter element can be arranged between the light source system and the sample to be tested, and is used to perform wavelength screening on the excitation light beam, so that only the excitation light beam of a specific wavelength is allowed to pass and reach the sample to be tested, so as to match the excitation peak of the fluorescent marker of the sample to be tested, that is, when the fluorescent substance is irradiated with light of a certain energy, the peak value of the light energy absorbed by the molecules in the process of transitioning from the ground state to the excited state appears at a specific wavelength. The second filter element can be arranged between the detector and the sample to be tested, and is used to perform wavelength screening on the optical signal to be detected generated by the sample to be tested, so as to screen out the light of a specific wavelength emitted by the fluorescent marker and exclude the interference of other non-specific light. For example, when measuring the absorbance of a solution of a sample to be tested, such as determining the concentration of protein or nucleic acid, according to the required measurement wavelength (such as 280nm for protein measurement), a filter element is set to allow only light of that wavelength to pass through. If multiple wavelengths need to be measured simultaneously, a switchable multi-wavelength filter system can be used (the module is set to switch automatically). For another example, in experiments for simultaneous detection of multiple markers, such as flow cytometry or cell imaging with multiple fluorescent markers, multiple filter channels are set according to the emission wavelengths of different fluorescent markers, and each channel corresponds to a specific wavelength range to achieve simultaneous detection of multiple fluorescent signals.

[0089] Therefore, the first filter element and the second filter element can adopt filters or multi-wavelength filter systems. In some embodiments, the wavelength filtering channels of the filters are preferably 455nm-505nm and 595nm-635nm. The range of 455nm-505nm usually corresponds to blue-green light, and the range of 595nm-635nm usually corresponds to orange to red light. Blue-green light and orange to red light correspond to the best excitation and emission wavelengths of different fluorescent dyes, respectively, which can ensure that the fluorescent signals between the dyes do not overlap, making it easier to distinguish and detect.

[0090] In some embodiments, the filter is preferably an infrared cutoff filter. Since the infrared cutoff filter can filter out infrared light and only allow visible light to pass, it can reduce the interference of non-target wavelengths of light (such as infrared light), thereby improving the signal-to-noise ratio and making the detection results more accurate and reliable.

[0091] In some embodiments, the optical detection device also includes a shutter element, which is arranged between the sample to be tested and the detector, and the switching time of the shutter element is less than 0.2S. By switching the switching state of the shutter element, the light signal to be detected of the sample to be tested can be controlled to reach the detector through the shutter element within a specific time. The switching time of the shutter element is less than 0.2s, and the shutter can be closed quickly and effectively, thereby achieving complete gated shading. In addition, the gating in the prior art usually uses a gating time window of the microsecond (μs) level, while the gating in the embodiment of the present invention can use a time window of the millisecond (ms) level to collect signals, and at the same time can measure the luminescence decay process with a duration of seconds (s). From microseconds to milliseconds, it means that the time resolution capability has been significantly improved, so that the luminescence process with a longer life (seconds) can be measured, which expands the scope of application.

[0092] The optical detection equipment of the present application is described in detail below with reference to specific embodiments. It should be noted that these examples do not constitute a limitation to the present application.

[0093] Embodiment 1:

[0094] like Figure 1As shown, in this embodiment, the light source system includes a first light source 100, a second light source 200 and a third light source 300, the spectroscopic element includes a first spectroscope 400, a second spectroscope 500, a third spectroscope 600, a fourth spectroscope 700 and a fifth spectroscope 800, and the detector includes a first detector 900, a second detector 1000 and a third detector 1100; wherein the first excitation light beam emitted by the first light source 100 is transmitted along the horizontal direction, the second excitation light beam emitted by the second light source 200 and the third excitation light beam emitted by the third light source 300 are both perpendicular to the first excitation light beam, and the second light source 200 and the third light source 300 are perpendicular to the first excitation light beam. The light sources 300 are staggered in the vertical direction, a first beam splitter 400 is arranged at the intersection of the first excitation beam and the second excitation beam, and a second beam splitter 500 is arranged at the intersection of the first excitation beam and the third excitation beam. The first beam splitter 400 and the second beam splitter 500 are used to convert the transmission direction of the second excitation beam and the third excitation beam from a vertical direction to a horizontal direction; the third beam splitter 600 is arranged between the light source system and the sample to be tested, and is used to reflect the first excitation beam and / or the second excitation beam and / or the third excitation beam to the sample to be tested; the wavelengths of the first excitation beam, the second excitation beam and the third excitation beam are different.

[0095] The three detectors can be PMT detectors, and filters are fixed at the front end of the detectors. The three filters can filter out light of three different wavelengths. When the light from the second light source 200 and the third light source 300 reaches the first spectroscope 400 and the second spectroscope 500, when the light enters the spectroscope at a specific angle (usually 45 degrees), the mirror surface will have a high transmittance or high reflectance for light of a specific wavelength, and show the opposite characteristics for light of other wavelengths. In this way, the transmission direction of the excitation light beam changes from vertical to parallel after passing through the spectroscope. When it reaches the intersection of the vertical direction with the first detector 900, a third spectroscope 600 is provided. The third spectroscope 600 reflects the light beam. The light is projected onto the sample to be tested below in the vertical direction. The sample to be tested will emit a light signal to be detected when stimulated by the light. After the light signal to be detected passes through the third beam splitter 600, it reaches the fourth beam splitter 700 above. Here, the feedback light beam is split into two paths, one of which passes through the fourth beam splitter 700 to reach the first detector 900, and the other is reflected in the horizontal direction. The fifth beam splitter 800 in the horizontal direction splits the light into two paths again, one of which reaches the second detector 1000 and the other reaches the third detector 1100. Since different filters are provided at the front ends of the first detector 900, the second detector 1000, and the third detector 1100, light of different wavelengths can be filtered out. Therefore, the wavelengths of light received by different detectors are actually different, so that information of three wavelengths can be obtained simultaneously in a single detection.

[0096] Embodiment 2:

[0097] like Figure 2 As shown, in this embodiment, different from Embodiment 1, two detectors are provided, one of which is a PMT detector 50 and the other is an APD detector 20, and a switchable filter system is installed between the PMT and APD detectors. In this example, the light source system includes a first laser 1 and a second laser 2. The light emitted by the first laser 1 is transmitted in a parallel direction, and the light of the second laser 2 is perpendicular to the parallel light beam of the first laser 1. When it reaches the intersection, it is reflected by the dichroic mirror 3 (beam splitter), and the light changes from the original vertical direction to a parallel direction. When it reaches the dichroic mirror 4, the light is reflected again, and excites the sample to be tested 6 to emit a light signal to be detected, which can be called emitted light. The emitted light passes through the dichroic mirror 4 and reaches the shutter element 7. The shutter element 7 can block the emitted light here. The PMT detector and the APD detector are normally open. In this way, when detection is required, the shutter element 7 is opened, and the emitted light reaches the dichroic mirror 8 above the shutter element 7 and is divided into two paths of light. One path passes through the dichroic mirror 8 and the filter 30 to reach the PMT detector 50, and the other path passes through the dichroic mirror 8 and the filter 9 to reach the APD detector 20. Here, since the installed filter is a switchable filter system, such as the IR-CUT dual filter switcher, an infrared cut-off filter is used to filter out infrared light to ensure the true restoration of the color image. When the light is insufficient, it switches to full-spectrum optical glass, allowing the detector to utilize all light and improve the detection capability under low-light conditions.

[0098] In some embodiments, the optical detection device further includes a focusing lens 5 and / or a collecting lens 10 and a collecting lens 40. The focusing lens 5 is arranged between the spectroscopic element and the sample to be tested, and the sample to be tested is located at the focal position of the focusing lens 5; the collecting lens 10 and the collecting lens 40 are arranged between the sample to be tested and the detector, and are used to collect the optical signal to be detected. The focusing lens 5 is fixed by an optical bracket to ensure that it is aligned with the optical axis between the dichroic mirror 4 and the sample to be tested 6. The dichroic mirror 4 here is used to divide the excitation light beam into two parts, one part is reflected to the focusing lens 5, so that the focusing lens 5 focuses the parallel light beam to the sample to be tested 6 to form a small light spot, and the other part of the light is used as a reference signal. In this way, the distribution and focusing of the light beam can be accurately controlled to ensure that the size and position of the light spot on the sample meet the detection requirements. The collecting lens 10 and the collecting lens 40 are used to collect the fluorescence emitted by the sample to be tested 6 so that the fluorescence is focused on the detector.

[0099] On the other hand, Figure 3As shown, an embodiment of the present invention further proposes an optical detection system, which includes the optical detection device, detection platform 1200, multi-well plate 1300, oscillation module (not shown), incubation module (not shown), heating module (not shown) and control module 1400 described in the above-mentioned embodiments; the multi-well plate 1300 is arranged on the detection platform 1200, the multi-well plate 1300 is used to carry the sample to be tested, the oscillation module is used to oscillate and mix the sample to be tested, and the incubation module is used to incubate the sample to be tested; the heating module is used to control the temperature of the incubation module; the control module 1400 is respectively communicated with the optical detection device, the detection platform 1200, the oscillation module, the incubation module and the heating module.

[0100] like Figure 4 As shown, in this example, the multi-well plate 1300 can be a 96-well plate, etc., for carrying the sample to be tested; the heating module includes a temperature compensation unit and a temperature sensing unit, wherein the temperature sensing unit is used to detect the temperature in real time, the temperature control accuracy of the temperature compensation unit is preferably ±0.2°C, and the inter-well range of the temperature compensation unit is not greater than 0.3°C. The temperature compensation unit includes a temperature compensation algorithm for correcting the output of the control module 1400. The control module 1400 is used to control the state of each module in the system, such as controlling the opening or closing of the light source system; setting the light source excitation time; temperature control of the heating module; mode control of the oscillation module. The control module 1400 reads the multi-well plate 1300 in at least one of the following modes: sequential reading, hole-by-hole reading, and custom reading. In some embodiments, the system further includes: a memory configured to store a computer program; a processor configured to execute a computer program.

[0101] In the above scheme, the porous plate 1300 and the aforementioned optical detection equipment are cleverly integrated together, and the optical detection equipment performs optical detection on the samples to be tested in the porous plate 1300. The design of the porous plate 1300 makes it possible to process multiple groups of samples simultaneously in a smaller space. It can be understood that the incubation module is used to incubate the samples to be tested at a specific temperature, the heating module is used to heat the samples to be tested, and the oscillation module is used to oscillate and mix the samples to be tested. In the entire system, the control module can control the temperature of the heating module, the working mode of the oscillation module such as the oscillation speed, oscillation time, etc., to meet the needs of different experiments. The design of this optical detection system enables the laboratory to efficiently carry out large-scale sample detection.

[0102] The content of this application is introduced below with specific embodiments. It should be noted that these examples do not constitute a limitation to this application.

[0103] Embodiment 3:

[0104] In order to overcome the limitations of existing detection technology in parallel detection of multiple targets, this embodiment provides an optical detection device for multi-channel parallel detection, the main feature of which is that it has multiple independent channels, each of which can independently perform light excitation and light reception, thereby realizing synchronous and efficient detection of multiple targets. This design has multiple independent detection channels, each of which can process different samples or targets in parallel, greatly improving the throughput and efficiency of detection. Each channel can be independently optimized according to the specific optical properties of the target to be detected (such as absorption wavelength, emission wavelength, fluorescence lifetime, etc.). This targeted optimization includes but is not limited to selecting a suitable excitation light source, filter, detector, etc. to ensure that each channel can detect a specific target with the highest efficiency. The system can increase or decrease the number of channels as needed to adapt to different detection requirements. Since each channel can work independently, the detection of multiple targets can be carried out simultaneously, greatly shortening the detection time. Independent optimization for each target helps to improve the accuracy and sensitivity of detection and reduce cross-interference. The system design takes into account different types of targets and detection methods, and can be compatible with multiple detection modes, such as fluorescence, phosphorescence, chemiluminescence, etc.

[0105] Each channel is equipped with an independent excitation light source, which can be a laser, LED or other type of light source. The appropriate excitation light wavelength is selected according to the absorption characteristics of the target. Each channel is equipped with an independent detector, such as a photomultiplier tube (PMT), avalanche photodiode (APD) or complementary metal oxide semiconductor (CMOS) sensor, which is used to receive and record the light signal emitted by the target. The advanced optical design ensures the spatial separation of the excitation light and the emission light to reduce optical crosstalk. Equipped with intelligent control software, it can adjust the parameters of each channel in real time, including light source intensity, detector gain, etc., to achieve precise control of the detection process.

[0106] In this embodiment, the optical detection device for multi-channel parallel detection includes a laser module, an optical path modulation module, a sample illumination and fluorescence excitation module, and a fluorescence collection and detection module.

[0107] In the laser module, it includes a laser and a collimating lens. The laser is located in front of the collimating lens to ensure that the excitation beam is directly irradiated on the collimating lens. The laser is fixed by an adjustable mounting seat, which is convenient for fine-tuning the position and angle of the laser. The collimating lens is installed in front of the laser and fixed by a precise optical bracket to ensure the alignment of the optical axis. The laser in this embodiment includes two solid-state lasers, for example, with wavelengths of 685nm and 730nm respectively. The collimating lens in this embodiment is used to convert the divergent light beam emitted by the laser into parallel light. In this way, a high-quality collimated beam can be achieved to ensure that the laser can still maintain a small spot size after long-distance transmission, thereby improving the detection sensitivity. Secondly, in the optical path modulation module, it includes a beam splitter and a focusing lens. The beam splitter is located after the collimating lens, and the focusing lens is located between the beam splitter and the sample. The beam splitter is fixed by a precise optical bracket to ensure that its angle and position are accurate. The focusing lens is also fixed by an optical bracket to ensure that it is aligned with the optical axis between the beam splitter and the sample. The beam splitter here is used to divide the laser beam into two parts, one part directly irradiates the sample, and the other part is used as a reference signal. The focusing lens focuses the parallel light beam onto the sample to form a small light spot. In this way, the distribution and focusing of the light beam can be precisely controlled to ensure that the size and position of the light spot on the sample meet the detection requirements. In the sample irradiation and fluorescence excitation module, it includes a sample stage and a fluorescence excitation filter. The sample stage is used to place the sample, usually a 96-well plate. The fluorescence excitation filter is located between the sample and the focusing lens, allowing only light of a specific wavelength to pass through. The sample stage is located at the focal position of the focusing lens, and the fluorescence excitation filter is located between the focusing lens and the sample. The sample stage is fixed by a precision mechanical structure to ensure the repeatability and stability of the sample position. The fluorescence excitation filter is fixed by an adjustable optical bracket, which is convenient for adjusting its position and angle. In this way, it can be ensured that only light of a specific wavelength can excite the fluorescent substance in the sample, thereby improving the specificity and sensitivity of fluorescence detection. Finally, the fluorescence collection and detection module includes a collection lens, an emission filter and an APD / PMT detector, wherein the collection lens collects the fluorescence emitted by the sample, the emission filter is located behind the collection lens, and only allows fluorescence of a specific wavelength to pass through, and the APD / PMT detector is used to detect the fluorescence signal. The collection lens is located behind the sample, and the emission filter is located between the collection lens and the APD / PMT. The collection lens is fixed by an optical bracket to ensure that it is aligned with the optical axis of the sample and the emission filter. The emission filter is fixed by an adjustable optical bracket to facilitate adjustment of its position and angle. The APD / PMT is fixed by a precise optical bracket to ensure that its receiving area is aligned with the optical axis of the emission filter. In this way, the fluorescence collection and detection module can efficiently collect and detect the fluorescence emitted by the sample, and improve the purity of the signal and the accuracy of detection through the selective filtering of the filter.

[0108] The above scheme of this embodiment can achieve high-precision beam control, efficient fluorescence excitation and collection, and accurate signal detection by precisely controlling the assembly and connection of each module, and the entire optical path design can meet the high standards required for biological detection.

[0109] Embodiment 4:

[0110] In order to achieve high-efficiency and high-throughput multi-target detection, the present application proposes an optical detection system, which not only includes the above-mentioned optical detection equipment, but also integrates an incubation unit, a heating unit, an oscillation unit, a 96-well plate and a software control system to achieve efficient processing and analysis of biological samples.

[0111] In the optical detection system of this embodiment, the 96-well plate is located on the detection platform of the optical detection device, and is used to carry the sample to be tested. The incubation unit is located below the 96-well plate, and is used to incubate the sample to be tested. The heating unit and the incubation unit can be integrated together to control the temperature during the incubation process. The oscillation unit is located next to the 96-well plate, and is used to oscillate and mix the sample to be tested. The control module is a central processing unit, which is connected to the incubation unit, the heating unit, the oscillation unit, and the optical detection device through a communication interface to achieve control and coordination of the entire system.

[0112] During the test, first, the sample to be tested is loaded into the 96-well plate. The control module sends instructions to the incubation unit and the oscillation unit through the communication connection to start the incubation and mixing of the sample. The incubation unit maintains a constant temperature through the heating unit, while the oscillation unit mixes the sample according to the set mode and speed. While the sample is being processed, the control module sets the parameters of the optical detection device according to the preset detection parameters, including the wavelength of the excitation light source. After the incubation and mixing treatment is completed, the 96-well plate is automatically moved to the detection platform of the optical detection device. The excitation light source emits light of a specific wavelength and irradiates the sample in the 96-well plate through an optical microscope. The target in the sample generates a fluorescent signal under the excitation of light, which is received by the detector and converted into an electrical signal. The control module collects the electrical signal data generated by the detector, and performs real-time or subsequent data analysis through the built-in software to generate the test results.

[0113] It is understandable that the oscillation mode of the oscillation unit in this embodiment can be set to different intensities, such as strong, medium and weak modes. In the strong vibration mode, the reaction speed can be increased and the reaction time can be shortened. The medium vibration mode is suitable for most conventional experiments to ensure sufficient mixing of the sample to be tested and the reagent. The weak vibration intensity is suitable for experiments that are sensitive to vibration. Of course, the oscillation unit can also support users to customize the parameters of the oscillation unit, such as the oscillation amplitude and frequency.

[0114] The system adopts an integrated design, integrating sample processing and optical detection on the same platform, reducing the number of operation steps and improving detection efficiency. Through the software control system, the sample processing and detection are automated, reducing manual intervention and reducing operational errors. The 96-well plate design allows for simultaneous processing and detection of multiple samples, achieving high-throughput detection.

[0115] Embodiment 5:

[0116] In this embodiment, an example is given of a 96-well plate automatically moving on a detection platform of an optical detection system. It can be considered that under the control instruction or trigger instruction of the control module, the 96-well plate moves synchronously with the detection platform, and the door opening and closing and mechanical resetting can be realized.

[0117] Specifically, the detection platform is equipped with a motor-driven mobile platform that can carry a 96-well plate. The movement of the detection platform is controlled by a control module. In addition, it also includes a sensor for detecting the position of the 96-well plate and the state of the door, and a switch door mechanism for controlling the opening and closing of the door. For example, a stepper motor or a servo motor is installed on the detection platform, and the platform is driven to move along the X-axis and Y-axis directions through motor control. At the position feedback level, an encoder or a grating ruler is installed on the platform to feedback the position information of the platform in real time. The control module sends a movement instruction to the motor according to the preset program or the instruction input by the user, and drives the platform to move to the specified position. A mechanical or electric door mechanism is installed next to the detection platform, and a sensor is installed near the door. The control module sends instructions to open and close the door as needed, and controls the opening or closing of the door through the drive device of the door mechanism. A reset button or limit switch is also installed on the detection platform. When a reset is required, the control module sends a reset instruction, drives the motor to return the platform to the initial position, and triggers the reset mechanism at the same time to ensure that the platform is accurately reset.

[0118] It is understood that the control module has a built-in control algorithm to ensure that the platform can move to the specified position accurately according to the instructions. The built-in algorithm in the control module can also adjust the parameters of the motor control system according to the information fed back by the sensor to achieve more precise position control. In addition, the built-in algorithm also includes an exception handling program. When encountering a fault or abnormal situation, the system can stop the operation in time and issue an alarm.

[0119] On the other hand, the embodiment of the present invention also proposes an optical detection method, such as Figure 5 As shown, the optical detection method includes the following steps:

[0120] Step S100: emitting at least two excitation light beams of different wavelengths to a sample to be tested, wherein the sample to be tested includes a plurality of homogeneous system luminescent materials;

[0121] Step S200: receiving an optical signal to be detected from a sample to be detected within a plurality of preset gated time windows;

[0122] Step S300: converting the optical signal to be detected into an electrical signal;

[0123] Step S400: acquiring light intensity information of the optical signal to be detected within each gated time window according to the electrical signal;

[0124] Step S500: calculating the initial luminescence signal intensity of the luminescent material of each homogeneous system according to the light intensity information;

[0125] Step S600: Calculate the concentration of the target in the corresponding sample to be tested according to the initial luminescent signal intensity.

[0126] Specifically, at least two excitation light beams of different wavelengths are emitted through a light source system, and the excitation light beams of different wavelengths do not interfere with each other during transmission and / or are independent in the light beam space; then, the spectroscopic element reflects and transmits the excitation light beam, and transmits the reflected or transmitted excitation light beam to the sample to be tested, so as to excite the sample to be tested to generate a light signal to be detected; then, the shutter element is opened within a plurality of preset gated time windows, so that the detector receives the light signal to be detected within the gated time window; the shutter element is arranged between the detector and the sample to be tested; the detector converts the received light signal to be detected into an electrical signal, and sends the electrical signal to the control module; the control module obtains the light intensity information of the light signal to be detected within each gated time window according to the electrical signal, and calculates the initial luminescence signal intensity of each luminescent material according to the light intensity information; the control module calculates the concentration of each target molecule according to the initial luminescence signal intensity.

[0127] In some embodiments, the control module calculates the concentration of the target in the corresponding sample to be tested according to the light intensity information as follows:

[0128] First, the luminescence lifetime of various luminescent materials used to detect the sample to be tested is obtained;

[0129] Secondly, the gating time parameters during detection are determined according to the luminescence lifetime, and the parameters are input into the control module;

[0130] Finally, the intensity of the individual light signal of the original target object to be detected is calculated according to the intensity of the mixed light signal obtained by detection.

[0131] In some embodiments, the steps of calculating the intensity of the original individual optical signal of the target to be detected by the mixed optical signal intensity are as follows:

[0132] For n targets to be tested, there will be n groups of homogeneous reagents containing luminescent materials, corresponding to n luminescence lifetimes τ n , so according to τ n Determine n-1 gate times tn , corresponding to n-1 light intensity signal values ​​I1-I n-1 , establish n-1 light intensity signal values ​​and the original light intensity signal value I A The relationship between is obtained, and the original concentration of the target object to be measured is calculated through the relationship.

[0133] It should be noted that, generally speaking, the luminescence lifetime refers to the time required for the light intensity to decay to 1 / e of the initial light intensity I0, but the gating time window here can be selected within the range of the light intensity decaying to 0 based on the luminescence lifetime.

[0134] In this example, gated detection can be achieved by the luminescence lifetime of the sample to be tested. Since different types of luminescent materials have different luminescence durations (i.e., luminescence lifetimes) after excitation, this difference can be used to distinguish target molecules in materials or samples to be tested. In gated detection technology, by setting a specific gated time window, only the luminescent signal detected within this gated time window belongs to the signal intensity of a specific molecule.

[0135] See also Figure 6 and Figure 7 , we can see that, assuming that there are three target molecules in the sample to be tested, there are three luminescence lifetimes. First, under the irradiation of the excitation light of different wavelengths emitted by the first laser, the second laser, and the third laser, the excitation light beam is transmitted to the sample to be tested. The sample to be tested is excited by the external light source and begins to emit light. After the excitation stops, the luminescent material will continue to emit light for a period of time, that is, Figure 7 In t1, t2, and t3, since the optical detection device is provided with a shutter element, this shutter element can be opened within a specific gated time window, and within the corresponding time window, the detector receives the luminescence signal. For example, within the time window from 0 to t1, the intensity of the detected light signal is the mixed light intensity I of the three luminescent materials. (1+2+3) In the time window of t1 to t2, the intensity of the detected light signal is the mixed light intensity I of the two luminescent materials with longer luminescence lifetimes. (2+3) Similarly, in the time window of t2 to t3, the intensity of the detected light signal is the light intensity I of the luminescent material with the longest luminescence lifetime. (3) Generally speaking, the photon number decay of luminescent materials is exponential decay, and there is the following formula:

[0136] I (t) =I0exp(-t / τ1);

[0137] Among them, I (t)It represents the luminescence signal intensity at time t, that is, the intensity of light emitted by the luminescent material at that moment; I0 ​​represents the initial luminescence signal intensity, that is, the light intensity of the luminescent material when it starts to emit light; exp(-t / τ1) is an attenuation factor, which is used to describe the attenuation of intensity as time t changes; τ1 is called the luminescence lifetime or attenuation constant, which is a parameter that characterizes the attenuation rate of the light intensity of the luminescent material, and represents the time required for the light intensity to decay to 1 / e of the initial light intensity I0. The longer the luminescence lifetime, the slower the luminescence decays of the material.

[0138] Therefore, based on the above formula, after obtaining the luminous signal intensity of the third material at a certain time between t2 and t3, the initial luminous signal intensity of the third material can be calculated according to the above formula. (2+3) , the initial luminescence signal intensity of the second material can be obtained, and so on, the initial luminescence signal intensity of the first material can be obtained.

[0139] Furthermore, a linear standard curve can be established between concentration and luminescence signal intensity (or photon number) and concentration. According to this standard curve, the actual initial luminescence signal intensity is substituted into the linear relationship to obtain the concentration of different target molecules in the sample to be tested.

[0140] It is understandable that the above method can also be used to calculate the number of different situations such as two target molecules, four target molecules, etc. in the sample to be tested. For example, assuming that there are two target molecules in the sample to be tested, in the time window of 0 to t1, the detected light signal intensity is the mixed light intensity I of the two luminescent materials. (1+2) In the time window of t1 to t2, the intensity of the detected light signal is the light intensity I of the luminescent material with the longest luminescence lifetime. (2) , the light intensity I of another luminescent material can be calculated (1) , and then using the above calculation method, the concentration of various target molecules can be calculated. Similarly, the same method can also be used to calculate the case where there are four or five target molecules.

[0141] It can be concluded from the above description that the optical detection method includes the following steps:

[0142] Detecting the luminescence intensity decay curves of n homogeneous systems of luminescent materials over time respectively, where n≥2 and n is a positive integer;

[0143] Select n time points t1, t2, ..., t n , respectively calculate the light intensity information of the luminescent material of each homogeneous system at these n time points; calculate the initial luminescent signal intensity of the luminescent material of each homogeneous system according to the light intensity information. It should be noted that the luminescent material is an afterglow material, and the structural formula of the luminescent material is as follows:

[0144]

[0145] Wherein, X1 and X2 are selected from one of S or O;

[0146] R1 and R2 are independently selected from at least one of the following groups:

[0147] (a) absent; (b) hydrogen; (c) halogen; (d) amino; or (e) a derivative of amino.

[0148] Wherein, the derivative of amino group is at least one of N-alkylated amino group, N,N-dialkylated amino group, N-arylamino group and N,N-diarylamino group. N-arylamino group is selected from one of N-phenylamino group, N-(substituted phenyl)amino group and N-(condensed aryl)amino group. The substituent of N-(substituted phenyl)amino group is selected from at least one of halogen, alkyl, alkoxy, amino group and nitro group. The N-(condensed aryl)amino group is selected from naphthyl, anthracenyl or phenanthrenyl.

[0149] Embodiment 6:

[0150] (1) Preparation of detection reagents

[0151] Preparation of carrier microspheres: Carboxyl polystyrene hollow microspheres were dissolved in 100 mL of deionized water, and ultrasonicated to form a dispersed phase to obtain a solution of carrier microspheres.

[0152] Preparation of donor microspheres: The sensitizer SiPc was dispersed in 10 mL of tetrahydrofuran solution. After the solution was prepared, it was quickly added to the above solution containing the carrier microspheres, and then gradually heated to 50°C. After continuous stirring for 10 hours, it was centrifuged and washed twice with deionized water and ethanol solution. The microspheres were stored in deionized water to prepare SiPc donor microspheres, which were then stored at room temperature in a brown reagent bottle or other light-proof container for later use.

[0153] Preparation of acceptor microspheres: Mix the buffer OO-Bu and the luminescent agent Eu(TTA)3(phen), and use the same preparation process as the donor microspheres to replace the sensitizer SiPc with the buffer OO-Bu and the luminescent agent Eu(TTA)3(phen) to obtain the acceptor microspheres.

[0154] The process for homogeneous receptor microspheres with other lifespans is the same as above, except that the buffer body is replaced.

[0155] The structure of the cache body is preferably as follows:

[0156]

[0157] The normalized lifetime decay curve of the above-mentioned buffer as a luminescent material in dichloromethane solvent at room temperature is as follows: Figure 8 As shown:

[0158] Microsphere-antibody coupling: Taking PCT as an example, N-hydroxysulfosuccinimide was added to the donor microspheres and the acceptor microspheres respectively, and the mixture was shaken and mixed. Then, an equal volume of (1-ethyl-(3-dimethylaminopropyl)carbodiimide) was added and shaken. After the activated solution was centrifuged, the supernatant was discarded, 1 mL of 2-(N-morpholino)ethanesulfonic acid was added for ultrasonication, and antibodies were added to the ultrasonicated mixture (labeled at a ratio of microspheres: antibody = 15:1), wherein PCT-Ab1 antibody was added to the donor microspheres and PCT-Ab2 antibody was added to the acceptor microspheres. The mixture was mixed in the dark, centrifuged and the supernatant was discarded. 1 mL of 1% bovine serum albumin was added to the precipitate for blocking. The mixture was ultrasonically mixed in the dark, centrifuged and the supernatant was discarded. Then, borate buffer (0.05 M, pH 8.0) was added to the precipitate, and the mixture was dispersed by ultrasonication and stored.

[0159] (2) Detection and calculation process

[0160] Homogeneous system mixing: In a three-channel homogenizer, set the temperature to 37°C. After the temperature stabilizes, open the chamber door, add 100 μL of test solution R1, 20 μL of the analyte PCT, and 30 μL of the donor microsphere solution coupled to the antibody to the 96-well plate in sequence, close the chamber door, and incubate with shaking for 2 minutes. Then add 30 μL of the receptor microsphere solution coupled to the antibody to the system, close the chamber door, and incubate with shaking for 8 minutes.

[0161] Experimental process for obtaining the long afterglow lifetime decay curve and k value: Taking the two-lifetime system as an example, the system is excited by 680nm excitation light, and the light signal at 615nm is collected. The excitation time is set to 0.5s, the delay time is 20ms, the single-point collection time is 0.1s, and the luminous intensity is continuously collected within 60s to obtain the intensity decay curve. The appropriate t1 and t2 are obtained by calculation and the k value is obtained.

[0162] Luminescence signal collection: Taking the two-lifetime system as an example, the system is excited by 680nm excitation light, and the light signal at 615nm is collected. Set the excitation time to 0.5s, the gate time t1=1s, t2=3s, and obtain the luminescence intensities I1 and I2 respectively. Substituting them into the calculation formula in the method, the luminescence intensity signal corresponding to the detection of different analytes can be restored, and the concentration of the analyte can be deduced using the working curve to achieve simultaneous detection of two analytes.

[0163] For example, for the detection of only two homogeneous systems, the luminescence intensity decay curves of homogeneous system A and homogeneous system B over time are experimentally detected. By selecting appropriate t1 and t2, the ratios of luminescence intensity at time t1 and t2, k1 and k2, can be obtained from the two decay curves respectively. According to the decay curve of the long afterglow system lifetime, it can be known that the ratio is a constant value related to the lifetime of the system, which is used to subsequently restore the luminescence intensity information of a single system when the two systems are mixed. The principle is as follows: Fig. 9 As shown:

[0164]

[0165] A homogeneous system A and a homogeneous system B with different lifetimes are mixed, and signals are collected at time t1 and t2 respectively after excitation to obtain I1 and I2:

[0166]

[0167] According to the above two formulas, the initial luminous intensity can be calculated:

[0168]

[0169] The schematic diagram of its principle is as follows Fig.10 shown.

[0170] After selecting t1 and t2, set the gate and collect the signal after a delay of t1 after the excitation light is turned off.

[0171] Taking the inflammatory markers PCT and IL-6 as examples, the luminescence intensity-analyte concentration working curve was established:

[0172] In the homogeneous system of buffer + antigen + left ball + right ball, the antigen concentrations were 0, 1, 2, 5, 10, and 20 ng / mL, respectively. After incubation, the cells were irradiated with excitation light, and multiple groups of luminescence intensity signals were collected at time t1. The luminescence intensity values ​​were plotted against the corresponding PCT concentrations to obtain a working curve. The working curve is shown in FIG. Fig.11 As shown:

[0173] According to the above-mentioned idea of ​​lifetime decoding, the accurate concentrations of the corresponding analytes A and B can be obtained through the standard curve and the measured I(A, t1) and I(B, t1). Similarly, the concentration of the target analyte in the multi-homogeneous system can be obtained.

[0174] In some embodiments, the optical detection method further comprises the following steps:

[0175] The light intensity sensing module detects the output intensity of the excitation light beams of different wavelengths of the light source system and sends the detection results to the processing module;

[0176] The processing module analyzes whether the output intensity of each excitation light beam deviates from the preset light source intensity according to the detection result, and sends a corresponding adjustment signal to the light source controller according to the excitation light beam with deviation;

[0177] The light source controller adjusts the output intensity of the corresponding excitation light beam of the light source system according to the adjustment signal.

[0178] Specifically, assuming that the light source system includes multiple light sources, if the stability of the light source is not high, the accuracy of the measurement result may be affected. To this end, a light intensity sensing module is integrated in the optical detection device. This module can use a highly sensitive photoelectric sensor to accurately capture the slight changes in the intensity of the light source. The light intensity sensing module is used to detect the output intensity of each wavelength light source in real time. For example, the light intensity sensing module is installed at the proximal end of each light source, that is, the initial part of the path of the light source emitting light, but does not affect the path of the light transmitted to the sample. A fixed point is set in the light path, and the light of all light sources must pass through this point. The light intensity sensing module is installed at this point, which can be used to monitor the light intensity passing through this point. The processing module may include an analog-to-digital converter and a processor. The processor may use an MCU, a CPU, a DSP, etc. The light intensity sensing module transmits the real-time monitoring data to the analog-to-digital converter, converts the analog signal of the light intensity into a digital signal; the digital signal is then transmitted to the processor, and the processor analyzes the data and compares it with the preset light source intensity. If the processor detects that the actual light source intensity deviates from the preset light source intensity, it will send an adjustment signal to the corresponding light source controller through the control interface. The light source controller corresponds to the light source one by one. The light source controller adjusts the output of the light source according to the adjustment signal received, such as adjusting the current or changing the driving parameters of the light source to correct the light intensity. Before the optical detection equipment starts detection, each light source is excited in turn to generate an excitation beam. The photoelectric sensor monitors the light intensity of each light source in real time. The light intensity data is transmitted to the processor in real time, and the processor compares the actual light intensity with the preset light intensity. If the intensity of any light source is lower than or higher than the predetermined range, the processor instructs the light source controller to make adjustments, such as increasing or decreasing the current. After adjustment, the light source is detected again until its intensity is adjusted to an appropriate level. This process ensures that the light source intensity is within the optimal range for each detection, thereby ensuring the accuracy and repeatability of the detection results.

[0179] In some embodiments, the optical detection method further comprises the following steps:

[0180] When the deviation between the output intensity of the excitation light beam of the light source and the preset light source intensity exceeds a threshold, the fuzzy logic controller switches the light source to a corresponding standby light source;

[0181] The fuzzy logic controller dynamically adjusts the switching speed and response time according to the real-time status of the light source system and system requirements.

[0182] Specifically, if the light intensity sensor module detects that the deviation exceeds the threshold, it means that there may be a problem with the light source. In this case, a redundant light source component is designed. The redundant light source component uses multi-threaded parallel processing technology so that the system can switch to the backup light source in time when a problem with the light source is detected. The fuzzy logic controller is used to dynamically adjust the switching speed and response time according to the real-time status of the light source and system requirements to ensure the continuity and accuracy of the detection.

[0183] For example, each wavelength of the light source in the light source system is equipped with at least one backup light source of the same wavelength. The backup light source is configured in parallel with the main light source, but is in standby mode under normal circumstances. Each pair of light sources (main light source and backup light source) is connected to a light source controller, which is responsible for managing the output and switching of the light sources. Multi-threaded parallel processing technology is used to monitor the status of all light sources at the same time. Each light source (including the backup light source) has an independent monitoring thread, which is responsible for collecting light intensity data and status information in real time. This design ensures the high responsiveness and real-time performance of the system to the light source status, so that it can quickly switch to the backup light source when there is a problem with the main light source. The integrated fuzzy logic controller is used to process the switching logic of the light source. The fuzzy logic controller receives data from the light intensity sensing module and evaluates the health status of the light source according to the preset rules and fuzzy logic algorithm. The controller dynamically adjusts the switching speed and response time to optimize the continuity and accuracy of the detection process, taking into account the real-time status of the light source and the detection requirements of the system. When the controller determines that the main light source needs to be replaced, it sends a switching signal to the light source controller. The light source controller immediately deactivates the main light source and starts the backup light source. The entire switching process can be completed in milliseconds. The switching mechanism also includes a feedback link. After the switching is completed, the new light source state is re-evaluated to ensure its normal operation. In summary, according to the optical detection equipment, system and method of the embodiments of the present application, it is possible to simultaneously or sequentially emit excitation light beams of different wavelengths, thereby realizing multi-wavelength detection of the sample to be tested. By partially reflecting and partially transmitting the incident light through the spectroscopic element, the intensity of the light signal to be detected received by the detector can be effectively improved, thereby enhancing the detection sensitivity and improving the reliability of the signal. In addition, the function of the spectroscopic element can filter out unnecessary light signals and only allow light of a specific wavelength to pass, thereby reducing interference from light signals of other wavelengths. The detection device includes multiple detectors, each of which can correspond to different wavelengths, so that multi-channel simultaneous detection can be achieved, greatly improving the detection efficiency.

[0184] On the other hand, an embodiment of the present invention further provides an electronic device, including:

[0185] A memory for storing program instructions;

[0186] The processor is used to call the program instructions stored in the memory and execute the above optical detection method according to the obtained program instructions.

[0187] The processor may be implemented by a general-purpose central processing unit (CPU), a microprocessor, an application-specific integrated circuit (ASIC), or one or more integrated circuits, and is used to execute relevant programs to implement the technical solutions provided in the embodiments of the present application;

[0188] The memory can be implemented in the form of a read-only memory (ROM), a static storage device, a dynamic storage device, or a random access memory (RAM). The memory can store an operating system and other application programs. When the technical solution provided in the embodiments of this specification is implemented by software or firmware, the relevant program code is stored in the memory, and the processor calls and executes the optical detection method of the embodiments of this application; the memory and the processor can be connected through a bus or the like.

[0189] On the other hand, an embodiment of the present invention further provides a storage medium, which is a computer-readable storage medium and stores a computer program. When the computer program is executed by a processor, the above-mentioned optical detection method is implemented.

[0190] As a non-transient computer-readable storage medium, the memory can be used to store non-transient software programs and non-transient computer executable programs. In addition, the memory may include a high-speed random access memory, and may also include a non-transient memory, such as at least one disk storage device, a flash memory device, or other non-transient solid-state storage devices. In some embodiments, the memory may optionally include a memory remotely arranged relative to the processor, and these remote memories may be connected to the processor via a network. Examples of the above-mentioned networks include, but are not limited to, the Internet, an intranet, a local area network, a mobile communication network, and a combination thereof. The device embodiments described above are merely schematic, wherein the units described as separate components may or may not be physically separated, and are implemented to be located in one place, or may also be distributed to multiple network units. Some or all of the modules may be selected according to actual needs to achieve the purpose of the present embodiment.

[0191] Although specific embodiments are described herein, those of ordinary skill in the art will recognize that many other modifications or alternative embodiments are also within the scope of the present disclosure. For example, any of the functions and / or processing capabilities described in conjunction with a particular device or component may be performed by any other device or component. In addition, although various exemplary implementations and architectures have been described according to embodiments of the present disclosure, those of ordinary skill in the art will recognize that many other modifications to the exemplary implementations and architectures described herein are also within the scope of the present disclosure.

[0192] Some aspects of the present disclosure are described above with reference to the block diagram and flow chart of the system, method, system and / or computer program product according to exemplary embodiments.It should be understood that the combination of one or more blocks in the block diagram and flow chart and the blocks in the block diagram and flow chart can be realized by executing computer executable program instructions respectively.Equally, according to some embodiments, some blocks in the block diagram and flow chart may not need to be executed in the order shown, or may not need to be executed in full.In addition, additional components and / or operations beyond those components and / or operations shown in the blocks in the block diagram and flow chart may be present in certain embodiments.

[0193] Therefore, the blocks in the block diagrams and flow charts support the combination of devices for performing the specified functions, the combination of elements or steps for performing the specified functions, and the program instruction devices for performing the specified functions. It should also be understood that each block in the block diagrams and flow charts and the combination of blocks in the block diagrams and flow charts can be implemented by a special-purpose hardware computer system or a combination of special-purpose hardware and computer instructions that performs a specific function, element or step.

[0194] The program modules, applications, etc. described herein may include one or more software components, including, for example, software objects, methods, data structures, etc. Each such software component may include computer-executable instructions that, in response to execution, cause at least a portion of the functionality described herein (e.g., one or more operations of the illustrative methods described herein) to be performed.

[0195] Software components can be encoded with any of various programming languages. An exemplary programming language can be a low-level programming language, such as an assembly language associated with a specific hardware architecture and / or an operating system platform. The software component comprising assembly language instructions may need to be converted to executable machine code by an assembler before being executed by hardware architecture and / or platform. Another exemplary programming language can be a higher-level programming language, which can be transplanted across multiple architectures. The software component comprising a higher-level programming language may need to be converted to an intermediate representation by an interpreter or a compiler before execution. Other examples of programming languages ​​include but are not limited to macro languages, shells or command languages, job control languages, scripting languages, database queries or search languages ​​or report writing languages. In one or more exemplary embodiments, the software component comprising the instruction of one of the above-mentioned programming language examples can be directly executed by an operating system or other software components, without first being converted into another form.

[0196] Software components may be stored as files or other data storage structures. Software components of similar type or related functions may be stored together, such as in a specific directory, folder, or library. Software components may be static (e.g., preset or fixed) or dynamic (e.g., created or modified at execution time).

[0197] The embodiments of the present invention are described in detail above with reference to the accompanying drawings, but the present invention is not limited to the above embodiments, and various changes can be made within the knowledge scope of ordinary technicians in the relevant technical field without departing from the purpose of the present invention.

Claims

1. An optical detection method, characterized in that: The following steps are involved: Emitting at least two excitation light beams of different wavelengths to a sample to be tested, to excite the sample to be tested to generate a light signal to be detected, wherein the sample to be tested includes a plurality of homogeneous system luminescent materials; Receiving the optical signal to be detected from the sample to be detected within a plurality of preset gated time windows; Converting the optical signal to be detected into an electrical signal; Acquire light intensity information of the optical signal to be detected within each of the gated time windows according to the electrical signal; Calculate the initial luminescence signal intensity of the luminescent material of each homogeneous system according to the light intensity information; The concentration of the target in the sample to be tested is calculated according to the initial luminescent signal intensity.

2. The optical detection method according to claim 1, characterized in that: The time intervals of the plurality of preset gated time windows are set based on the luminescence lifetimes of the luminescent materials of the plurality of homogeneous systems.

3. The optical detection method according to claim 2, characterized in that: The step of acquiring the light intensity information of the light signal to be detected within each gated time window according to the electrical signal, and calculating the initial luminescence signal intensity of the luminescent material of each homogeneous system according to the light intensity information comprises: Detecting the luminescence intensity decay curves of n homogeneous systems of luminescent materials over time respectively, where n≥2 and n is a positive integer; Select n time points t1, t2, ..., t n , respectively calculate the light intensity information of the luminescent material of each homogeneous system at these n time points; and calculate the initial luminescent signal intensity of the luminescent material of each homogeneous system based on the light intensity information.

4. The optical detection method according to claim 1, characterized in that: The luminescent material is an afterglow material.

5. The optical detection method according to claim 4, characterized in that: The structural formula of the luminescent material is shown below: Wherein, X1 and X2 are selected from one of S or O; R1 and R2 are independently selected from at least one of the following groups: (a) absent; (b) hydrogen; (c) halogen; (d) amino; or (e) a derivative of amino.

6. The optical detection method according to claim 5, characterized in that: The amino derivative is at least one of an N-alkylated amino group, an N,N-dialkylated amino group, an N-arylamino group, and an N,N-diarylamino group.

7. The optical detection method according to claim 6, characterized in that: The N-arylamino group is selected from the group consisting of: N-phenylamino, N-(substituted phenyl)amino, and N-(condensed ring aryl)amino.

8. The optical detection method according to claim 7, characterized in that: The substituent of the N-(substituted phenyl)amino group is selected from at least one of halogen, alkyl, alkoxy, amino and nitro.

9. The optical detection method according to claim 7, characterized in that: The N-(fused ring aryl)amino group is selected from naphthyl, anthracenyl or phenanthrenyl.

10. An optical detection device, characterized in that: include: A light source system for emitting at least two excitation light beams of different wavelengths; the excitation light beams of different wavelengths do not interfere with each other during transmission and / or are independent in light beam space; A spectroscopic element, used for reflecting and transmitting the excitation light beam, so that the reflected or transmitted excitation light beam is transmitted to the sample to be tested, and excites the sample to be tested to generate a light signal to be detected; The detector is used to receive the optical signal to be detected and convert the optical signal to be detected into an electrical signal representing a detection result.

11. The optical detection device according to claim 10, characterized in that: The optical detection equipment also includes a light intensity sensing module, a processing module and a light source controller. The light intensity sensing module is used to detect the output intensity of the excitation light beams of different wavelengths of the light source system and send the detection results to the processing module; the processing module is used to analyze whether the output intensity of each excitation light beam deviates from the preset light source intensity, and send a corresponding adjustment signal to the light source controller based on the excitation light beam with the deviation, so that the light source controller adjusts the output intensity of the corresponding excitation light beam of the light source system.

12. The optical detection device according to claim 11, characterized in that: The light source system includes at least two light sources, and the at least two light sources are used to emit excitation light beams of at least two different wavelengths; the optical detection equipment also includes a fuzzy logic controller and at least two backup light sources, and the backup light sources correspond to the light sources one by one. When the deviation between the output intensity of the excitation light beam of the light source and the preset light source intensity exceeds a threshold, the fuzzy logic controller is used to switch the light source to the corresponding backup light source. The fuzzy logic controller is also used to dynamically adjust the switching speed and response time according to the real-time status of the light source system and system requirements.

13. The optical detection device according to claim 10, characterized in that: The optical detection device further comprises a collimating element, which is arranged at the output end of the light source system and is used for converting the excitation light beam into a parallel light beam.

14. The optical detection device according to claim 10, characterized in that: The optical detection device also includes a first filter element and / or a second filter element; the excitation light beam is transmitted to the sample to be tested through the first filter element, and the first filter element is used to perform wavelength filtering on the excitation light beam; the second filter element is arranged between the sample to be tested and the detector, and is used to perform wavelength filtering on the optical signal to be detected.

15. The optical detection device according to claim 10, characterized in that: The optical detection device further comprises a shutter element, which is arranged between the sample to be detected and the detector, and the on-off switching time of the shutter element is less than 0.2S.

16. The optical detection device according to claim 10, characterized in that: The light source system includes at least two light sources, and the at least two light sources are used to emit excitation light beams of at least two different wavelengths; or, the light source system includes a tunable laser, and the tunable laser can emit excitation light beams of at least two different wavelengths under different parameters.

17. The optical detection device according to claim 10, characterized in that: The spectroscopic element includes a first spectroscope, a second spectroscope and a third spectroscope; the light source system includes a first light source, a second light source and a third light source, wherein the first excitation light beam emitted by the first light source is transmitted in a horizontal direction, the second excitation light beam emitted by the second light source and the third excitation light beam emitted by the third light source are both perpendicular to the first excitation light beam, and the second light source and the third light source are staggered in a vertical direction, the first spectroscope is arranged at the intersection of the first excitation light beam and the second excitation light beam, the second spectroscope is arranged at the intersection of the first excitation light beam and the third excitation light beam, the first spectroscope and the second spectroscope are used to convert the transmission direction of the second excitation light beam and the third excitation light beam from a vertical direction to a horizontal direction; the third spectroscope is arranged between the light source system and the sample to be tested, and is used to reflect the first excitation light beam and / or the second excitation light beam and / or the third excitation light beam to the sample to be tested; the wavelengths of the first excitation light beam, the second excitation light beam and the third excitation light beam are different.

18. The optical detection device according to claim 17, characterized in that: The spectroscopic element further includes a fourth spectroscope and a fifth spectroscope; the detector includes a first detector, a second detector and a third detector, the fourth spectroscope is arranged between the first detector and the sample to be tested, and is used to transmit the optical signal to be detected to the first detector, and reflect the optical signal to be detected to the fifth spectroscope; The fifth spectroscope is used to reflect the light signal to be detected reflected by the first detector to the second detector, and to transmit the light signal to be detected reflected by the first detector to the third detector; the input ends of the first detector, the second detector and the third detector are all provided with filter elements.

19. The optical detection device according to claim 10, characterized in that: The optical detection device further comprises a focusing lens and / or a collecting lens, wherein the focusing lens is arranged between the light splitting element and the sample to be tested, and the sample to be tested is located at a focal position of the focusing lens; The collecting lens is disposed between the sample to be detected and the detector, and is used to collect the optical signal to be detected.

20. An optical detection system, characterized in that: include: The optical detection device according to any one of claims 10 to 19; Testing platform; A multi-well plate, disposed on the detection platform, and used for carrying samples to be tested; An oscillation module, used for oscillating and mixing the sample to be tested; An incubation module, used for incubating the sample to be tested; A heating module, used to control the temperature of the incubation module; The control module is respectively connected to the optical detection device, the detection platform, the oscillation module, the incubation module and the heating module.

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