Fluorescence characteristic curve calibration method and device for algae classification equipment

Abstract

The invention discloses a fluorescence characteristic curve calibration method and device for algae classification equipment, and the method comprises the steps: measuring fluorescence intensity signals of a standard fluorescent substance under different excitation wavelengths, and calculating a characteristic curve of a fluorescent substance solution through the fluorescence intensity signals under different excitation wavelengths and the concentration of the fluorescent substance; wherein the fluorescent substance concentration is obtained by preparing a standard fluorescent substance, and according to the characteristic curve of the fluorescent substance solution and a pre-established relation model, determining fluorescence characteristic curves of different types of algae; wherein the relation model stores the corresponding relation between the fluorescence standard substance and the algae fluorescence characteristic coefficient, and the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment is carried out through the fluorescence characteristic curves of different types of algae.

Description

Technical Field

[0001] The present application relates to the technical field of water quality detection, and more specifically, to a fluorescence characteristic curve calibration method and device for algae classification equipment. Background Art

[0002] Eutrophication of water bodies and the massive reproduction of algae in water are the main causes of red tides and algal blooms. It not only causes significant economic losses to humans, but also poses a major threat to health and even life. Therefore, real-time and effective detection of the existing amount of algae in water is an important part of water pollution detection and control.

[0003] In the prior art, algae classification monitors are used to detect the amount of algae in water and obtain the concentration of various algae (green algae, blue algae, diatoms, cryptophytes). Due to differences in equipment components and equipment manufacturing processes, each fluorescent algae classification device needs to be regularly measured and calibrated for fluorescence characteristic curves before leaving the factory and during use.

[0004] At present, the determination and calibration of fluorescence characteristic curves mainly adopt the living algae method. In the process of determining and calibrating the fluorescence characteristic curve using the living algae method, there are problems such as poor repeatability of different batches of algae solutions, large amount of algae solution used, and large measurement workload, which leads to poor stability and accuracy of the measurement results of the fluorescence algae classification equipment.

[0005] Therefore, how to improve the stability and accuracy of the measurement results of the fluorescence algae classification equipment is an urgent problem to be solved in this application. Summary of the invention

[0006] In view of this, the present application discloses a fluorescence characteristic curve calibration method and device for an algae classification device, aiming to improve the accuracy and stability of the fluorescence characteristic curve calibration of the fluorescence algae classification device.

[0007] In order to achieve the above purpose, the disclosed technical solution is as follows:

[0008] The first aspect of the present application discloses a method for calibrating a fluorescence characteristic curve of an algae classification device, the method comprising:

[0009] Measure the fluorescence intensity signal of standard fluorescent substances at different excitation wavelengths;

[0010] The characteristic curve of the fluorescent substance solution is calculated by the fluorescence intensity signal and the fluorescent substance concentration under the different excitation wavelengths; wherein the fluorescent substance concentration is obtained by preparing a standard fluorescent substance;

[0011] Determine the fluorescence characteristic curves of different types of algae according to the characteristic curve of the fluorescent substance solution and a pre-established relationship model; wherein the relationship model stores the corresponding relationship between the fluorescent standard substance and the fluorescence characteristic coefficient of the algae;

[0012] The fluorescence characteristic curve of the fluorescence algae classification device is calibrated by using the fluorescence characteristic curves of the different types of algae.

[0013] Preferably, the measuring of the fluorescence intensity signal of the standard fluorescent substance at different excitation wavelengths includes:

[0014] Excitation light is emitted to standard fluorescent substance solutions of multiple gradient concentrations to be detected through light-emitting diodes of different wavelengths, so that fluorescence intensity signals at different excitation wavelengths can be measured under the alternating action of high-frequency pulses of light-emitting diodes of different wavelengths.

[0015] Preferably, the characteristic curve of the fluorescent substance solution is calculated by using the fluorescence intensity signals and the fluorescent substance concentrations at the different excitation wavelengths, including:

[0016] Under the fluorescence intensity at different excitation wavelengths, the concentration of the fluorescent substance prepared by the standard fluorescent substance is taken as the independent variable, and the fluorescence intensity signal at different excitation wavelengths is taken as the dependent variable, and linear regression fitting is performed to obtain the fluorescence characteristic coefficient of the fluorescent standard substance;

[0017] The characteristic curve of the fluorescent substance solution is determined by the fluorescence characteristic coefficient of the fluorescent standard substance.

[0018] Preferably, determining the fluorescence characteristic curves of different types of algae according to the characteristic curve of the fluorescent substance solution and a pre-established relationship model comprises:

[0019] Determine the pure algae sample by detecting the chlorophyll concentration and identifying the species of the algae solution; wherein the pure algae at least include green algae, blue algae, diatoms and cryptoalgae;

[0020] Scanning the fluorescence spectrum of a water sample of a fixed-concentration fluorescence standard substance and a sample of the pure algae to establish a corresponding relationship between the fluorescence standard substance and the fluorescence characteristic coefficient of the algae in the relationship model;

[0021] The fluorescence characteristic curves of different types of algae are determined through the characteristic curve of the fluorescent substance solution and the corresponding relationship between the fluorescent standard substance and the algae fluorescence characteristic coefficient in the established relationship model.

[0022] Preferably, it also includes:

[0023] The chlorophyll concentration of the algae solution was measured by spectrophotometry, and the algae solution was identified by morphology and molecular biology to obtain algae solutions with multiple gradient concentrations.

[0024] Preferably, the step of calibrating the fluorescence characteristic curve of the fluorescence algae classification device by using the fluorescence characteristic curves of the different types of algae comprises:

[0025] The algae classification concentration of the fluorescent standard substance is re-measured through the fluorescence characteristic curves of the different types of algae to verify the calibration of the fluorescence characteristic curve of the fluorescence algae classification equipment.

[0026] Preferably, the process of establishing the relationship model includes:

[0027] Based on the characteristic curve of the fluorescent substance solution, the fluorescence characteristic curves of different types of algae are normalized to establish a relationship model of the corresponding relationship between the fluorescent standard substance and the fluorescence characteristic coefficient of the algae.

[0028] The second aspect of the present application discloses a fluorescence characteristic curve calibration device for algae classification equipment, the device comprising:

[0029] A measuring unit, used for measuring the fluorescence intensity signal of a standard fluorescent substance at different excitation wavelengths;

[0030] A calculation unit, used to calculate the characteristic curve of the fluorescent substance solution through the fluorescence intensity signal and the fluorescent substance concentration under the different excitation wavelengths; wherein the fluorescent substance concentration is obtained by preparing a standard fluorescent substance;

[0031] A determination unit, used to determine the fluorescence characteristic curves of different types of algae according to the characteristic curve of the fluorescent substance solution and a pre-established relationship model; wherein the relationship model stores the corresponding relationship between the fluorescent standard substance and the fluorescence characteristic coefficient of the algae;

[0032] The calibration unit is used to calibrate the fluorescence characteristic curve of the fluorescence algae classification device through the fluorescence characteristic curves of the different types of algae.

[0033] Preferably, the measuring unit is specifically used for:

[0034] Excitation light is emitted to standard fluorescent substance solutions of multiple gradient concentrations to be detected through light-emitting diodes of different wavelengths, so that fluorescence intensity signals at different excitation wavelengths can be measured under the alternating action of high-frequency pulses of light-emitting diodes of different wavelengths.

[0035] Preferably, the computing unit comprises:

[0036] A linear regression fitting module is used to perform linear regression fitting under different fluorescence intensities at different excitation wavelengths, taking the concentration of the fluorescent substance prepared by the standard fluorescent substance as an independent variable and the fluorescence intensity signal at different excitation wavelengths as a dependent variable, to obtain the fluorescence characteristic coefficient of the fluorescent standard substance;

[0037] The first determination module is used to determine the characteristic curve of the fluorescent substance solution according to the fluorescence characteristic coefficient of the fluorescent standard substance.

[0038] Through the above technical scheme, it can be known that the present application discloses a fluorescence characteristic curve calibration method and device for algae classification equipment, which measures the fluorescence intensity signal of the standard fluorescent substance at different excitation wavelengths, and calculates the characteristic curve of the fluorescent substance solution through the fluorescence intensity signal at different excitation wavelengths and the concentration of the fluorescent substance; wherein the concentration of the fluorescent substance is obtained by preparing the standard fluorescent substance, and the fluorescence characteristic curves of different types of algae are determined according to the characteristic curve of the fluorescent substance solution and the pre-established relationship model; wherein the relationship model stores the corresponding relationship between the fluorescent standard substance and the algae fluorescence characteristic coefficient, and the fluorescence characteristic curve of the fluorescence method algae classification equipment is calibrated through the fluorescence characteristic curves of different types of algae. Through the above scheme, since the current determination and calibration of the fluorescence characteristic curve mainly adopts the living algae method, the living algae method calibration process has the problems of large amount of algae solution used, large measurement workload, and poor stability of measurement results. Fluorescent substances are standardized reagents with strong stability and good batch repeatability. They can be traced to the source of measurement to ensure the accuracy of the fluorescence characteristic curve. Therefore, fluorescent standard substances are used instead of living algae. The characteristic curve of the fluorescent substance solution is calculated through the concentration of fluorescent substances prepared by standard fluorescent substances and the fluorescence intensity signals under different excitation wavelengths. The fluorescence characteristic curve of the fluorescent algae classification equipment is calibrated based on the characteristic curve of the fluorescent substance solution and the fluorescence characteristic curves of different types of algae determined by the pre-established relationship model, thereby improving the stability and accuracy of the fluorescence characteristic curve calibration of the fluorescent algae classification equipment. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying any creative work.

[0040] Figure 1 A schematic diagram of a flow chart of a fluorescence characteristic curve calibration method for an algae classification device disclosed in an embodiment of the present application;

[0041] Figure 2 This is a schematic diagram of calculating the characteristic curve of each solution disclosed in the examples of this application;

[0042] Figure 3 A schematic diagram of establishing the relationship between the fluorescence standard substance and the algae fluorescence characteristic curve disclosed in the embodiment of the present application;

[0043] Figure 4 A schematic flow chart of another method for calibrating a fluorescence characteristic curve of an algae classification device disclosed in an embodiment of the present application;

[0044] Figure 5 A schematic diagram of the structure of a fluorescence characteristic curve calibration device for an algae classification device disclosed in an embodiment of the present application;

[0045] Figure 6 This is a schematic diagram of the structure of an electronic device disclosed in an embodiment of the present application. DETAILED DESCRIPTION

[0046] The following will be combined with the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0047] In this application, the terms "comprises", "comprising" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device comprising a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, an element defined by the sentence "comprising a ..." does not exclude the presence of other identical elements in the process, method, article or device comprising the element.

[0048] As can be seen from the background technology, at present, the determination and calibration of the fluorescence characteristic curve mainly adopts the living algae method. In the process of measuring and calibrating the fluorescence characteristic curve by the living algae method, there are problems such as poor repeatability of different batches of algae solutions, large amount of algae solution used, and large measurement workload, which leads to poor stability and accuracy of the measurement results of the fluorescence algae classification equipment. Therefore, how to improve the stability and accuracy of the measurement results of the fluorescence algae classification equipment is a problem that needs to be solved urgently in this application.

[0049] In order to solve the above problems, the present application discloses a method and device for calibrating the fluorescence characteristic curve of an algae classification device. Since the current determination and calibration of the fluorescence characteristic curve mainly adopts the living algae method, the living algae method calibration process has the problems of large amount of algae solution used, large measurement workload, and poor stability of the measurement results. Fluorescent substances are standardized reagents with strong stability and good batch repeatability. They can trace the value and ensure the accuracy of the fluorescence characteristic curve. Therefore, fluorescent standard substances are used instead of living algae. The characteristic curve of the fluorescent substance solution is calculated by the concentration of the fluorescent substance prepared by the standard fluorescent substance and the fluorescence intensity signal under different excitation wavelengths. According to the characteristic curve of the fluorescent substance solution and the fluorescence characteristic curves of different types of algae determined by the pre-established relationship model, the fluorescence characteristic curve of the fluorescence method algae classification device is calibrated, thereby improving the stability and accuracy of the fluorescence characteristic curve calibration of the fluorescence method algae classification device. The specific implementation method is specifically described through the following embodiments.

[0050] refer to Figure 1 As shown, a fluorescence characteristic curve calibration method of an algae classification device disclosed in an embodiment of the present application is shown, and the fluorescence characteristic curve calibration method of the algae classification device mainly includes the following steps:

[0051] S101: Acquire fluorescence intensity signals of standard fluorescent substances at different excitation wavelengths.

[0052] In S101, light emitting diodes of different wavelengths emit excitation light to standard fluorescent substance solutions of multiple gradient concentrations to be detected, so that fluorescence intensity signals at different excitation wavelengths are obtained under the alternating action of high-frequency pulses of light measuring light from light emitting diodes of different wavelengths.

[0053] Fluorescence algae classification monitoring is mainly based on the algae excitation fluorescence spectrum classification measurement method based on multi-wavelength light emitting diodes (LEDs). When the same algae is excited by excitation light of different wavelengths and unit intensity, the fluorescence intensity emitted is different. When different algae are excited by excitation light of the same wavelength and unit intensity, the fluorescence intensity emitted is also different. The algae classification monitor emits multi-wavelength excitation light to the test water body through LEDs of different wavelengths. Under the alternating action of high-frequency pulses of LED measurement light of different wavelengths, the synchronous fluorescence intensity signal excited by various excitation lights is obtained. Combined with the built-in fluorescence characteristic curves of different algae in the monitor, the concentration of various algae (green algae, blue algae, diatoms, cryptophytes) can be obtained using the least squares method. Due to differences in the original components of the equipment and the manufacturing process of the equipment, each algae classification equipment needs to be calibrated and measured regularly for the fluorescence characteristic curve before leaving the factory and during use.

[0054] The standard fluorescent substance must have strong fluorescence characteristics at an excitation wavelength of 300-600nm and an emission wavelength of 610-750nm.

[0055] S102: Calculate a characteristic curve of the fluorescent substance solution through the fluorescence intensity signal and the fluorescent substance concentration at different excitation wavelengths; wherein the fluorescent substance concentration is obtained by preparing a standard fluorescent substance.

[0056] In S102, multiple fluorescent substance solutions with gradient concentrations are used, and excitation lights of different wavelengths are used to sequentially irradiate the fluorescent substance solutions with different concentrations. The fluorescence intensity generated by the solution under the irradiation of the excitation light is measured, and the linear relationship between the fluorescence intensity and the fluorescent substance concentration at different excitation wavelengths is calculated to obtain a characteristic curve of the fluorescent substance solution.

[0057] At present, the determination and calibration of fluorescence characteristic curves mainly adopt the living algae method. The calibration process of the living algae method has the problems of large amount of algae solution used, large measurement workload and poor stability of measurement results. Fluorescent substances are standardized reagents with strong stability and good batch repeatability. They can trace the value and ensure the accuracy of fluorescence characteristic curves. Therefore, fluorescent standard substances are used instead of living algae to calibrate the fluorescence characteristic curve of fluorescence algae classification equipment. Compared with the existing technology, this solution reduces the amount of reagents used by 75% and the calibration work time by 75%. It can trace the value and improve the accuracy of the fluorescence characteristic curve, thereby improving the stability and accuracy of the fluorescence characteristic curve calibration of the fluorescence algae classification equipment.

[0058] The specific process of calculating and obtaining the characteristic curve of the fluorescent substance solution is as described in A1-A2.

[0059] A1: Under different fluorescence intensities at different excitation wavelengths, the concentration of the fluorescent substance prepared by the standard fluorescent substance is taken as the independent variable, and the fluorescence intensity signal at different excitation wavelengths is taken as the dependent variable, and linear regression fitting is performed to obtain the fluorescence characteristic coefficient of the fluorescent standard substance.

[0060] A2: Determine the characteristic curve of the fluorescent substance solution through the fluorescence characteristic coefficient of the fluorescent standard substance.

[0061] This scheme uses fluorescent standard substances instead of living algae for the calibration of the fluorescence characteristic curve of the fluorescence algae classification equipment. Living algae have poor stability and batch repeatability, making it difficult to trace the value of the laboratory. Fluorescent substances are standardized reagents with strong stability and good batch repeatability. They can be traced and ensure the accuracy of the fluorescence characteristic curve.

[0062] The characteristic curve of the fluorescent substance solution is as follows Figure 2 shown.

[0063] Figure 2 In the figure, the horizontal axis represents the excitation wavelength in nanometers (nm); the vertical axis represents the fluorescence characteristic coefficient.

[0064] S103: Determine the fluorescence characteristic curves of different types of algae according to the characteristic curve of the fluorescent substance solution and a pre-established relationship model; wherein the relationship model stores the corresponding relationship between the fluorescent standard substance and the fluorescence characteristic coefficient of the algae.

[0065] In S103, the characteristic curve of the fluorescent substance solution is used as a reference to normalize the fluorescence characteristic curves of different types of algae, so as to establish a relationship model of the corresponding relationship between the fluorescent standard substance and the fluorescence characteristic coefficient of the algae.

[0066] Due to the influence of the installation and manufacturing process of light sources, lenses, detectors, etc., spectral equipment still has certain differences between each device, even if it is the same model. Therefore, the normalization of the fluorescence characteristic curves of different types of algae is to establish the relationship between the fluorescence standard material and the fluorescence characteristic curve of algae, and eliminate the influence of factors such as different instrument responses on the fluorescence intensity. It is also to enable the relationship model between the fluorescence standard material and the fluorescence characteristic curve of algae to be migrated and applied between different devices.

[0067] Specifically, the process of determining the fluorescence characteristic curves of different types of algae based on the characteristic curve of the fluorescent substance solution and the pre-established relationship model is shown in B1-B3.

[0068] B1: Determine the pure algae sample by detecting the chlorophyll concentration and identifying the species of the algae solution; wherein the pure algae at least include green algae, blue algae, diatoms and cryptoalgae.

[0069] B2: Scan the fluorescence spectra of water samples of fixed-concentration fluorescent standard substances and pure algae samples to establish the corresponding relationship between the fluorescent standard substances and the algae fluorescence characteristic coefficients in the relationship model.

[0070] Specifically, four pure algae samples (green algae, blue algae, diatoms, and cryptoalgae) were determined through laboratory testing of chlorophyll a concentration and morphological / molecular biology identification of species. Water samples with fixed concentrations of fluorescent standard substances and the four pure algae samples were subjected to fluorescence spectral scanning to establish a corresponding relationship between the fluorescent standard substances and the algae fluorescence characteristic coefficients.

[0071] B3: Determine the fluorescence characteristic curves of different types of algae through the characteristic curve of the fluorescent substance solution and the correspondence between the fluorescent standard substance and the algae fluorescence characteristic coefficient in the pre-established relationship model.

[0072] The characteristic curve of the fluorescent substance solution is obtained, and the relationship model between the fluorescent standard substance and the algae fluorescence characteristic curve established above is used to calculate the fluorescence characteristic curves of different types of algae.

[0073] The corresponding relationship between the fluorescence standard substance and the algae fluorescence characteristic coefficient is shown in Table 1.

[0074] The specific relationship between the fluorescence standard substance and the algae fluorescence characteristic coefficient can be seen in Table 1.

[0075] Table 1

[0076]

[0077] In Table 1, green algae solutions with concentrations of 7.32ug / L and 14.36ug / L, blue algae solutions with concentrations of 13.77ug / L and 27.48ug / L, diatom solutions with concentrations of 6.79ug / L and 13.87ug / L, cryptoalgae solutions with concentrations of 10.03ug / L and 19.71ug / L, and fluorescent substance solutions with concentrations of 20ug / L and 40ug / L were prepared, and multi-wavelength excitation light (365nm, 410nm, 440nm, 470nm, 530nm, 570nm, 590nm) was sequentially incident on the water samples to measure the fluorescence intensity generated by the solutions under the irradiation of the excitation light.

[0078] Determine the fluorescence characteristic curves of different algae species such as Figure 3 shown. Figure 3 A schematic diagram showing the relationship between the fluorescent standard substance and the algae fluorescence characteristic curve is shown.

[0079] Figure 3 In the figure, the horizontal axis represents the excitation wavelength in nanometers (nm); the vertical axis represents the normalized fluorescence characteristic coefficient.

[0080] S104: Calibrate the fluorescence characteristic curve of the fluorescence algae classification device using the fluorescence characteristic curves of different algae species.

[0081] In S104, the algae classification concentration of the fluorescent standard substance is re-measured through the fluorescence characteristic curves of different types of algae to verify the calibration of the fluorescence characteristic curve of the fluorescence algae classification device.

[0082] Fluorescence standard materials were retested using the fluorescence characteristic curves of different algae species to evaluate and calibrate the accuracy of algae classification equipment.

[0083] Adjust the settings of the fluorescence algae classification equipment according to the retest results to ensure that the algae classification equipment can accurately classify and quantify different algae species based on the fluorescence characteristic curve.

[0084] Calibrated equipment is validated using independent algae species samples to ensure calibration validity and equipment reliability.

[0085] Specifically, the algae classification concentration of the fluorescent standard substance is re-measured through the fluorescence characteristic curves of different types of algae to verify the process of calibration of the fluorescence characteristic curve of the fluorescence algae classification equipment, which is explained in conjunction with Table 2.

[0086] Table 2

[0087]

[0088] In Table 2, fluorescent substance solutions with concentrations of 20ug / L and 40ug / L were prepared, and multi-wavelength excitation light (365nm, 410nm, 440nm, 470nm, 530nm, 570nm, 590nm) was sequentially incident on the water sample to measure the fluorescence intensity of the solution under the irradiation of the excitation light. The characteristic curve of the fluorescent substance solution was obtained, and the relationship between the fluorescence characteristic curve of the fluorescent standard substance and the algae fluorescence characteristic curve established above was used to calculate the fluorescence characteristic curves of different types of algae. The algae classification concentration of the fluorescent standard substance was re-measured through the fluorescence characteristic curves of different types of algae to verify the calibration of the fluorescence characteristic curve of the fluorescence algae classification equipment.

[0089] This scheme uses fluorescent standard substances instead of living algae for the calibration of the fluorescence characteristic curve of the fluorescence algae classification equipment. Specifically, the fluorescent substance needs to have strong fluorescence characteristics at an excitation wavelength of 300-600nm and an emission wavelength of 610-750nm.

[0090] The fluorescence characteristic curves of different types of algae are shown in Table 3.

[0091] Table 3

[0092]

[0093] In order to facilitate the understanding of the fluorescence characteristic curve calibration process of the algae classification equipment, combined with Figure 4 Let's take an example:

[0094] Figure 4 In the process, the fixed concentration of fluorescent standard material water sample is determined through material certificate and concentration traceability calculator;

[0095] Through laboratory testing of chlorophyll a concentration and morphological / molecular biological species identification, four pure algae samples (green algae, blue algae, diatoms, and cryptophytes) were determined;

[0096] The water samples with fixed concentration of fluorescent standard substances and four kinds of pure algae samples were subjected to fluorescence spectrum scanning to establish the relationship between the fluorescence standard substances and the fluorescence characteristic coefficients of algae.

[0097] Prepare gradient concentrations of standard fluorescent substances;

[0098] The fluorescence intensity under different excitation wavelengths was detected by an algae classification analyzer;

[0099] Under the fluorescence intensity at different excitation wavelengths, the fluorescence characteristic coefficient of the fluorescence standard substance is calculated; specifically, the concentration of the fluorescent substance prepared by the standard fluorescent substance is used as the independent variable, and the fluorescence intensity signal at different excitation wavelengths is used as the dependent variable, and linear regression fitting is performed to obtain the fluorescence characteristic coefficient of the fluorescence standard substance;

[0100] The fluorescence characteristic curves of different algae species are determined by the fluorescence characteristic coefficients of different algae species, the characteristic curves of the fluorescent substance solution and the corresponding relationship between the fluorescence standard substance and the fluorescence characteristic coefficients of the algae in the pre-established relationship model;

[0101] The algae classification concentration (ie, actual measured value) of the fluorescent standard substance is re-measured through the fluorescence characteristic curves of different types of algae to verify the calibration of the fluorescence characteristic curve of the fluorescence algae classification equipment.

[0102] This scheme uses multiple gradient concentration fluorescent substance solutions, and excitation lights of different wavelengths sequentially illuminate fluorescent substance solutions of different concentrations, measures the fluorescence intensity generated by the solution under the irradiation of the excitation light, calculates the linear relationship between the fluorescence intensity and the concentration of the fluorescent substance at different excitation wavelengths, and obtains the characteristic curve of the fluorescent substance solution; uses the obtained characteristic curve of the fluorescent substance solution, and the relationship between the fluorescent standard substance and the algae fluorescence characteristic curve to calculate the fluorescence characteristic curve of different types of algae, and completes the fluorescence characteristic curve calibration of the fluorescence algae classification equipment. Specifically, the method for establishing the relationship between the fluorescent standard substance and the algae fluorescence characteristic curve is: using multiple gradient concentration algae solutions and fluorescent substance solutions, and excitation lights of different wavelengths sequentially illuminate algae solutions and fluorescent substance solutions of different concentrations, measures the fluorescence intensity generated by the solution under the irradiation of the excitation light, calculates the linear relationship between the fluorescence intensity and the concentration of algae or fluorescent substance at different excitation wavelengths, obtains the characteristic curves of the algae solution and the fluorescent substance solution, and establishes the relationship between the fluorescent standard substance and the algae fluorescence characteristic curve.

[0103] In the embodiment of the present application, since the determination and calibration of the fluorescence characteristic curve currently mainly adopts the living algae method, the living algae method calibration process has the problems of large amount of algae solution used, large measurement workload, and poor stability of the measurement results. Fluorescent substances are standardized reagents with strong stability and good batch repeatability, and can be traced to the source of the value to ensure the accuracy of the fluorescence characteristic curve. Therefore, fluorescent standard substances are used instead of living algae, and the characteristic curve of the fluorescent substance solution is calculated by the fluorescent substance concentration prepared by the standard fluorescent substance and the fluorescence intensity signal under different excitation wavelengths. According to the characteristic curve of the fluorescent substance solution and the fluorescence characteristic curves of different types of algae determined by the pre-established relationship model, the fluorescence characteristic curve of the fluorescence method algae classification device is calibrated, thereby improving the stability and accuracy of the fluorescence characteristic curve calibration of the fluorescence method algae classification device.

[0104] Based on the above embodiments Figure 1 A fluorescence characteristic curve calibration method for an algae classification device is disclosed. The embodiment of the present application also discloses a fluorescence characteristic curve calibration device for an algae classification device, such as Figure 5 As shown, the fluorescence characteristic curve calibration device of the algae classification device includes:

[0105] A measuring unit 501 is used to measure the fluorescence intensity signal of the standard fluorescent substance at different excitation wavelengths;

[0106] The calculation unit 502 is used to calculate the characteristic curve of the fluorescent substance solution through the fluorescence intensity signal and the fluorescent substance concentration under different excitation wavelengths; wherein the fluorescent substance concentration is obtained by preparing a standard fluorescent substance;

[0107] The determination unit 503 is used to determine the fluorescence characteristic curves of different types of algae according to the characteristic curve of the fluorescent substance solution and the pre-established relationship model; wherein the relationship model stores the corresponding relationship between the fluorescent standard substance and the fluorescence characteristic coefficient of the algae;

[0108] The calibration unit 504 is used to calibrate the fluorescence characteristic curve of the fluorescence algae classification device by using the fluorescence characteristic curves of different types of algae.

[0109] Furthermore, the measuring unit 501 is specifically used to emit excitation light to the standard fluorescent substance solutions of multiple gradient concentrations to be detected through light-emitting diodes of different wavelengths, so that under the alternating action of high-frequency pulses of light-emitting diodes of different wavelengths, fluorescence intensity signals at different excitation wavelengths can be measured.

[0110] Furthermore, the calculation unit 502 includes:

[0111] A linear regression fitting module is used to perform linear regression fitting under different fluorescence intensities at different excitation wavelengths, taking the concentration of the fluorescent substance prepared by the standard fluorescent substance as an independent variable and the fluorescence intensity signal at different excitation wavelengths as a dependent variable, to obtain the fluorescence characteristic coefficient of the fluorescent standard substance;

[0112] The first determination module is used to determine the characteristic curve of the fluorescent substance solution according to the fluorescence characteristic coefficient of the fluorescent standard substance.

[0113] Further, the determining unit 503 includes:

[0114] The second determination module is used to determine the pure algae sample by detecting the chlorophyll concentration and the species obtained by identifying the algae solution; wherein the pure algae at least include green algae, blue algae, diatoms and cryptoalgae;

[0115] A scanning module is used to perform fluorescence spectrum scanning on a water sample of a fixed-concentration fluorescent standard substance and a sample of the pure algae, so as to establish a corresponding relationship between the fluorescent standard substance and the algae fluorescence characteristic coefficient in the relationship model;

[0116] The third determination module is used to determine the fluorescence characteristic curves of different types of algae through the characteristic curve of the fluorescent substance solution and the corresponding relationship between the fluorescent standard substance and the algae fluorescence characteristic coefficient in the established relationship model.

[0117] Furthermore, the fluorescence characteristic curve calibration device of the algae classification device also includes:

[0118] The identification unit is used to measure the chlorophyll concentration of the algae solution by spectrophotometry, and to identify the algae solution by morphology and molecular biology to obtain algae solutions with multiple gradient concentrations.

[0119] Furthermore, the calibration unit 504 is specifically used to re-measure the algae classification concentration of the fluorescent standard substance through the fluorescence characteristic curves of different types of algae to verify the calibration of the fluorescence characteristic curve of the fluorescence algae classification device.

[0120] Furthermore, the determination unit 503 for establishing a relationship model is specifically used to normalize the fluorescence characteristic curves of different types of algae based on the characteristic curve of the fluorescent substance solution to establish a relationship model of the corresponding relationship between the fluorescent standard substance and the fluorescence characteristic coefficient of the algae.

[0121] In the embodiment of the present application, since the determination and calibration of the fluorescence characteristic curve currently mainly adopts the living algae method, the living algae method calibration process has the problems of large amount of algae solution used, large measurement workload, and poor stability of the measurement results. Fluorescent substances are standardized reagents with strong stability and good batch repeatability, and can be traced to the source of the value to ensure the accuracy of the fluorescence characteristic curve. Therefore, fluorescent standard substances are used instead of living algae, and the characteristic curve of the fluorescent substance solution is calculated by the fluorescent substance concentration prepared by the standard fluorescent substance and the fluorescence intensity signal under different excitation wavelengths. According to the characteristic curve of the fluorescent substance solution and the fluorescence characteristic curves of different types of algae determined by the pre-established relationship model, the fluorescence characteristic curve of the fluorescence method algae classification device is calibrated, thereby improving the stability and accuracy of the fluorescence characteristic curve calibration of the fluorescence method algae classification device.

[0122] An embodiment of the present application further provides a storage medium, which includes stored instructions, wherein when the instructions are executed, the device where the storage medium is located is controlled to execute the fluorescence characteristic curve calibration method for the algae classification device as described above.

[0123] The present application also provides an electronic device, whose structural diagram is as follows: Figure 6As shown, it specifically includes a memory 601, and one or more instructions 602, wherein the one or more instructions 602 are stored in the memory 601, and are configured to be executed by one or more processors 603 to execute the fluorescence characteristic curve calibration method of the above-mentioned algae classification equipment.

[0124] For the aforementioned method embodiments, for the sake of simplicity, they are all described as a series of action combinations, but those skilled in the art should be aware that the present application is not limited by the order of the actions described, because according to the present application, some steps can be performed in other orders or simultaneously. Secondly, those skilled in the art should also be aware that the embodiments described in the specification are all preferred embodiments, and the actions and modules involved are not necessarily required by the present application.

[0125] It should be noted that each embodiment in this specification is described in a progressive manner, and each embodiment focuses on the differences from other embodiments, and the same or similar parts between the embodiments can be referred to each other. For the device embodiment, since it is basically similar to the method embodiment, the description is relatively simple, and the relevant parts can be referred to the partial description of the method embodiment.

[0126] The steps in the methods of the embodiments of the present application can be adjusted in order, combined, and deleted according to actual needs.

[0127] Finally, it should be noted that, in this article, relational terms such as first and second, etc. are merely used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations.

[0128] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present application. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the scope of the present application. Therefore, the present application will not be limited to the embodiments shown herein, but will conform to the widest scope consistent with the principles and novel features disclosed herein.

[0129] The above is only a preferred implementation of the present application. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present application. These improvements and modifications should also be regarded as the scope of protection of the present application.

Claims

1. A method for calibrating the fluorescence characteristic curve of an algae classification device, characterized in that: The method comprises: Measure the fluorescence intensity signal of standard fluorescent substances at different excitation wavelengths; The characteristic curve of the fluorescent substance solution is calculated by the fluorescence intensity signal and the fluorescent substance concentration under the different excitation wavelengths; wherein the fluorescent substance concentration is obtained by preparing a standard fluorescent substance; Determine the fluorescence characteristic curves of different types of algae according to the characteristic curve of the fluorescent substance solution and a pre-established relationship model; wherein the relationship model stores the corresponding relationship between the fluorescent standard substance and the fluorescence characteristic coefficient of the algae; The fluorescence characteristic curve of the fluorescence algae classification device is calibrated by using the fluorescence characteristic curves of the different types of algae.

2. The method according to claim 1, characterized in that The method of measuring the fluorescence intensity signal of the standard fluorescent substance at different excitation wavelengths includes: Excitation light is emitted to standard fluorescent substance solutions of multiple gradient concentrations to be detected through light-emitting diodes of different wavelengths, so that fluorescence intensity signals at different excitation wavelengths can be measured under the alternating action of high-frequency pulses of light-emitting diodes of different wavelengths.

3. The method according to claim 1, characterized in that The step of calculating the characteristic curve of the fluorescent substance solution by using the fluorescence intensity signals and the fluorescent substance concentrations at the different excitation wavelengths comprises: Under the fluorescence intensity at different excitation wavelengths, the concentration of the fluorescent substance prepared by the standard fluorescent substance is taken as the independent variable, and the fluorescence intensity signal at different excitation wavelengths is taken as the dependent variable, and linear regression fitting is performed to obtain the fluorescence characteristic coefficient of the fluorescent standard substance; The characteristic curve of the fluorescent substance solution is determined by the fluorescence characteristic coefficient of the fluorescent standard substance.

4. The method according to claim 1, characterized in that: Determining the fluorescence characteristic curves of different types of algae according to the characteristic curve of the fluorescent substance solution and a pre-established relationship model includes: Determine the pure algae sample by detecting the chlorophyll concentration and identifying the species of the algae solution; wherein the pure algae at least include green algae, blue algae, diatoms and cryptoalgae; Scanning the fluorescence spectrum of a water sample of a fixed-concentration fluorescence standard substance and a sample of the pure algae to establish a corresponding relationship between the fluorescence standard substance and the fluorescence characteristic coefficient of the algae in the relationship model; The fluorescence characteristic curves of different types of algae are determined through the characteristic curve of the fluorescent substance solution and the corresponding relationship between the fluorescent standard substance and the algae fluorescence characteristic coefficient in the established relationship model.

5. The method according to claim 4, characterized in that Also includes: The chlorophyll concentration of the algae solution was measured by spectrophotometry, and the algae solution was identified by morphology and molecular biology to obtain algae solutions with multiple gradient concentrations.

6. The method according to claim 1, characterized in that The step of calibrating the fluorescence characteristic curve of the fluorescence algae classification device by using the fluorescence characteristic curves of the different types of algae includes: The algae classification concentration of the fluorescent standard substance is re-measured through the fluorescence characteristic curves of the different types of algae to verify the calibration of the fluorescence characteristic curve of the fluorescence algae classification equipment.

7. The method according to claim 1, characterized in that The process of building a relationship model includes: Based on the characteristic curve of the fluorescent substance solution, the fluorescence characteristic curves of different types of algae are normalized to establish a relationship model of the corresponding relationship between the fluorescent standard substance and the fluorescence characteristic coefficient of the algae.

8. A fluorescence characteristic curve calibration device for algae classification equipment, characterized in that: The device comprises: A measuring unit, used for measuring the fluorescence intensity signal of the standard fluorescent substance at different excitation wavelengths; A calculation unit, used to calculate the characteristic curve of the fluorescent substance solution through the fluorescence intensity signal and the fluorescent substance concentration under the different excitation wavelengths; wherein the fluorescent substance concentration is obtained by preparing a standard fluorescent substance; A determination unit, used to determine the fluorescence characteristic curves of different types of algae according to the characteristic curve of the fluorescent substance solution and a pre-established relationship model; wherein the relationship model stores the corresponding relationship between the fluorescent standard substance and the fluorescence characteristic coefficient of the algae; The calibration unit is used to calibrate the fluorescence characteristic curve of the fluorescence algae classification device through the fluorescence characteristic curves of the different types of algae.

9. The device according to claim 8, characterized in that The measuring unit is specifically used for: Excitation light is emitted to standard fluorescent substance solutions of multiple gradient concentrations to be detected through light-emitting diodes of different wavelengths, so that fluorescence intensity signals at different excitation wavelengths can be measured under the alternating action of high-frequency pulses of light-emitting diodes of different wavelengths.

10. The device according to claim 8, characterized in that The computing unit comprises: A linear regression fitting module is used to perform linear regression fitting under different fluorescence intensities at different excitation wavelengths, taking the concentration of the fluorescent substance prepared by the standard fluorescent substance as an independent variable and the fluorescence intensity signal at different excitation wavelengths as a dependent variable, to obtain the fluorescence characteristic coefficient of the fluorescent standard substance; The first determination module is used to determine the characteristic curve of the fluorescent substance solution according to the fluorescence characteristic coefficient of the fluorescent standard substance.

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