A fluorescence characteristic curve calibration method and device of an algae classification device

Abstract

The application discloses a fluorescence characteristic curve calibration method and device of an algae classification equipment, measures fluorescence intensity signals of standard fluorescent substances under different excitation wavelengths, and calculates characteristic curves of the fluorescent substance solutions by the fluorescence intensity signals under the different excitation wavelengths and fluorescent substance concentrations. The fluorescent substance concentrations are obtained by preparing the standard fluorescent substances, the fluorescence characteristic curves of different kinds of algae are determined according to the characteristic curves of the fluorescent substance solutions and a pre-established relationship model. The relationship model stores a corresponding relationship between the fluorescent standard substances and algae fluorescence characteristic coefficients, and the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment is performed by the fluorescence characteristic curves of the different kinds of algae.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of water quality detection, and more particularly to a fluorescence characteristic curve calibration method and device of an algae classification device. BACKGROUND

[0002] Water eutrophication and the massive reproduction of algae in water are the main causes of red tide and water bloom. It not only causes great 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 treatment.

[0003] In the prior art, the existing amount of algae in water is detected by an algae classification monitor, and the concentrations of various algae (green algae, blue-green algae, diatoms, and cryptophytes) are obtained. Due to differences in device components and manufacturing processes, the fluorescence characteristic curve of each fluorescence method algae classification device needs to be measured and calibrated regularly before it is shipped and during its use.

[0004] Currently, the fluorescence characteristic curve is mainly measured and calibrated using live algae. The process of measuring and calibrating the fluorescence characteristic curve using live algae has the problems of poor repeatability of different batches of algae solution, large use of algae solution, and large measurement workload, which leads to poor stability and accuracy of the measurement results of the fluorescence method algae classification device.

[0005] Therefore, how to improve the stability and accuracy of the measurement results of the fluorescence method algae classification device is a problem that needs to be solved by the present application. SUMMARY

[0006] Therefore, the present application discloses a fluorescence characteristic curve calibration method and device of an algae classification device, which aims to improve the accuracy and stability of the fluorescence characteristic curve calibration of the fluorescence method algae classification device.

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

[0008] The first aspect of the present application discloses a fluorescence characteristic curve calibration method of an algae classification device, which comprises:

[0009] measuring the fluorescence intensity signals of a standard fluorescent substance under different excitation wavelengths;

[0010] calculating the characteristic curve of the fluorescent substance solution from the fluorescence intensity signals under the different excitation wavelengths and the concentration of the fluorescent substance, wherein the concentration of the fluorescent substance is prepared by a standard fluorescent substance;

[0011] According to the characteristic curve of the fluorescent substance solution and the pre-established relationship model, the fluorescent characteristic curves of different kinds of algae are determined; wherein, the corresponding relationship between the fluorescent standard substance and the algae fluorescent characteristic coefficient is stored in the relationship model;

[0012] Through the fluorescent characteristic curves of the different kinds of algae, the fluorescent characteristic curve calibration of the fluorescent method algae classification equipment is performed.

[0013] Preferably, the measurement of the fluorescent intensity signal of the standard fluorescent substance under different excitation wavelengths comprises:

[0014] Through the different wavelengths of the light-emitting diode, the excitation light is emitted to the standard fluorescent substance solution with multiple gradient concentrations to be detected, so that the fluorescent intensity signal under different excitation wavelengths is measured under the alternating action of the high-frequency pulse of the light-emitting diode measurement light of different wavelengths.

[0015] Preferably, the calculation of the characteristic curve of the fluorescent substance solution through the fluorescent intensity signal under different excitation wavelengths and the fluorescent substance concentration comprises:

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

[0017] Through the fluorescent characteristic coefficient of the fluorescent standard substance, the characteristic curve of the fluorescent substance solution is determined.

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

[0019] Through the detected chlorophyll concentration and the kind obtained by identifying the algae solution, the pure algae sample is determined; wherein, the pure algae at least includes green algae, blue-green algae, diatom and cryptomonad;

[0020] The sample of the pure algae and the fixed-concentration fluorescent standard substance water sample are subjected to fluorescent spectrum scanning to establish the corresponding relationship between the fluorescent standard substance and the algae fluorescent characteristic coefficient in the relationship model;

[0021] Through the characteristic curve of the fluorescent substance solution and the corresponding relationship between the fluorescent standard substance and the algae fluorescent characteristic coefficient in the established relationship model, the fluorescent characteristic curves of different kinds of algae are determined.

[0022] Preferably, it further comprises:

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

[0024] Preferably, the fluorescence characteristic curve calibration of the fluorescence method algal classification device through the fluorescence characteristic curves of the different kinds of algae comprises:

[0025] The algal classification concentration of the fluorescence standard substance is re-measured through the fluorescence characteristic curves of the different kinds of algae to verify the fluorescence characteristic curve calibration of the fluorescence method algal classification device.

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

[0027] The fluorescence characteristic curves of the different kinds of algae are normalized based on the characteristic curve of the fluorescence substance solution to establish a relationship model of the corresponding relationship between the fluorescence standard substance and the algal fluorescence characteristic coefficient.

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

[0029] A measurement unit for measuring the fluorescence intensity signals of the standard fluorescence substance under different excitation wavelengths;

[0030] A calculation unit for calculating the characteristic curve of the fluorescence substance solution through the fluorescence intensity signals under the different excitation wavelengths and the fluorescence substance concentration, wherein the fluorescence substance concentration is prepared by the standard fluorescence substance;

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

[0032] A calibration unit for performing the fluorescence characteristic curve calibration of the fluorescence method algal classification device through the fluorescence characteristic curves of the different kinds of algae.

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

[0034] The standard fluorescence substance solution with multiple gradient concentrations to be detected is excited by light emitting diodes of different wavelengths to make the fluorescence intensity signals under different excitation wavelengths be measured under the alternating action of high-frequency pulses of the light emitting diodes of different wavelengths.

[0035] Preferably, the calculation unit comprises:

[0036] A linear regression fitting module for performing linear regression fitting by taking the fluorescence substance concentration prepared by the standard fluorescence substance as the independent variable and taking the fluorescence intensity signals under the different excitation wavelengths as the dependent variable to obtain the fluorescence characteristic coefficient of the fluorescence standard substance.

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

[0038] According to the technical solution, the fluorescence intensity signals of the standard fluorescent substance under different excitation wavelengths are measured, the characteristic curve of the fluorescent substance solution is calculated based on the fluorescence intensity signals under different excitation wavelengths and the concentration of the fluorescent substance, the concentration of the fluorescent substance is prepared by using the standard fluorescent substance, the fluorescence characteristic curves of different types of algae are determined based on the characteristic curve of the fluorescent substance solution and the pre-established relationship model, the corresponding relationship between the fluorescent standard substance and the algae fluorescence characteristic coefficient is stored in the relationship model, and the fluorescence characteristic curve calibration of the fluorescence algae classification equipment is performed based on the fluorescence characteristic curves of different types of algae. In the above solution, the fluorescence characteristic curve is mainly calibrated by using the living algae method, and the living algae method has the problems of large consumption of algae solution, large measurement workload, and poor stability of measurement results. The fluorescent substance is a standardized reagent, has strong stability, good batch repeatability, and can be traced to the value, and thus the accuracy of the fluorescence characteristic curve is ensured. Therefore, the living algae is replaced by the fluorescent standard substance, the concentration of the fluorescent substance prepared by using the standard fluorescent substance and the fluorescence intensity signals under different excitation wavelengths are used to calculate the characteristic curve of the fluorescent substance solution, the fluorescence characteristic curves of different types of algae are determined based on the characteristic curve of the fluorescent substance solution and the pre-established relationship model, and the fluorescence characteristic curve calibration of the fluorescence algae classification equipment is performed, so that the stability and accuracy of the fluorescence characteristic curve calibration of the fluorescence algae classification equipment are improved. BRIEF DESCRIPTION OF DRAWINGS

[0039] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed in the embodiments or the prior art description will be briefly introduced. Obviously, the drawings in the following description are only embodiments of the present application, and those skilled in the art can obtain other drawings according to the provided drawings without creative labor.

[0040] Figure 1 A flowchart of a fluorescence characteristic curve calibration method of an algae classification equipment disclosed in an embodiment of the present application is shown in the figure.

[0041] Figure 2 A schematic diagram of calculating the characteristic curves of each solution disclosed in an embodiment of the present application is shown in the figure.

[0042] Figure 3 A schematic diagram of establishing the relationship between the fluorescent standard substance and the algae fluorescence characteristic curve disclosed in an embodiment of the present application is shown in the figure.

[0043] Figure 4 Another flowchart of a method for calibrating a fluorescence characteristic curve of an algae classification device according to an embodiment of the present application is disclosed.

[0044] Figure 5 Another structural diagram of a device for calibrating a fluorescence characteristic curve of an algae classification device according to an embodiment of the present application is disclosed.

[0045] Figure 6 Another structural diagram of an electronic device according to an embodiment of the present application is disclosed. DETAILED DESCRIPTION

[0046] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of protection of the present application.

[0047] In the present application, the term “comprises”, “comprising”, or any other variant thereof is intended to cover non-exclusive inclusion, so that a process, method, article, or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or further includes elements inherent to such a process, method, article, or device. Without more limitations, the element defined by the statement “comprises a” does not exclude the presence of another identical element in the process, method, article, or device including the element.

[0048] As known from the background, at present, the determination and calibration of the fluorescence characteristic curve mainly adopts the living algae method. In the process of determining and calibrating the fluorescence characteristic curve by the living algae method, there are problems such as poor repeatability of different batches of algae solution, large amount of algae solution used, and large amount of measurement work, which leads to poor stability and accuracy of the measurement results of the fluorescence algae classification device. Therefore, how to improve the stability and accuracy of the measurement results of the fluorescence algae classification device is a problem to be solved by the present application.

[0049] In order to solve the above problems, the application discloses a fluorescence characteristic curve calibration method and device of an algae classification equipment. Since the determination and calibration of the fluorescence characteristic curve currently mainly adopts the living algae method, there are large amounts of algae solution, large measurement workload and poor stability of the measurement results in the calibration process of the living algae method. The fluorescence substance is a standardized reagent, has strong stability, good batch repeatability, can perform value tracing, and ensures the accuracy of the fluorescence characteristic curve. Therefore, the living algae is replaced by the fluorescence standard substance, the fluorescence substance concentration and the fluorescence intensity signal under different excitation wavelengths prepared by the standard fluorescence substance are calculated to obtain the characteristic curve of the fluorescence substance solution, the fluorescence characteristic curves of different types of algae are determined according to the characteristic curve of the fluorescence substance solution and the relationship model established in advance, the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment is performed, and the stability and accuracy of the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment are improved. The specific implementation manner is specifically explained through the following embodiments.

[0050] Reference Figure 1 As shown in the figure, the algae classification equipment fluorescence characteristic curve calibration method disclosed by the application mainly includes the following steps:

[0051] S101: Obtain the fluorescence intensity signal of the standard fluorescence substance under different excitation wavelengths.

[0052] In S101, the standard fluorescence substance solution with multiple gradient concentrations to be detected is irradiated with excitation light through light-emitting diodes (LEDs) of different wavelengths, so that the fluorescence intensity signal under different excitation wavelengths is obtained under the alternating action of the high-frequency pulse of the measurement light of the LEDs of different wavelengths.

[0053] The fluorescence method algae classification monitoring is mainly a multi-wavelength LED-based algae excitation fluorescence spectrum classification measurement method. When the same type of algae is excited by excitation light of different wavelengths and unit intensities, 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 water body to be detected through LEDs of different wavelengths, obtains the synchronous fluorescence intensity signal excited by various excitation light under the alternating action of the high-frequency pulse of the measurement light of the LEDs of different wavelengths, and then combines the fluorescence characteristic curves of different algae built in the monitor to obtain the concentration of each type of algae (green algae, blue-green algae, diatoms, and cryptophytes) by using the least square method. Due to the differences in the original devices and the manufacturing process of the equipment, each algae classification equipment needs to be calibrated and determined for the fluorescence characteristic curve before leaving the factory and during use.

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

[0055] S102: Obtain the characteristic curve of the fluorescent substance solution by calculating the fluorescent intensity signals under different excitation wavelengths and the concentration of the fluorescent substance; wherein the concentration of the fluorescent substance is obtained by preparing a standard fluorescent substance.

[0056] In S102, a plurality of gradient concentrations of the fluorescent substance solution are used, different wavelengths of excitation light are sequentially irradiated on different concentrations of the fluorescent substance solution, the fluorescence intensity generated by the solution under the irradiation of the excitation light is measured, the linear relationship between the fluorescence intensity under different excitation wavelengths and the concentration of the fluorescent substance is calculated, and the characteristic curve of the fluorescent substance solution is obtained.

[0057] Since the determination and calibration of the fluorescence characteristic curve at present mainly adopts the live algae method, there are large amount of algae solution used, large amount of measurement work, and poor stability of measurement results in the calibration process of the live algae method. The fluorescent substance is a standardized reagent, has strong stability, good batch repeatability, can perform value traceability, and ensures the accuracy of the fluorescence characteristic curve. Therefore, the fluorescent standard substance is used to replace the live algae for the calibration of the fluorescence characteristic curve of the fluorescence method algae classification equipment. Compared with the prior art, the reagent usage is reduced by 75%, the calibration work time is reduced by 75%, value traceability can be performed, the accuracy of the fluorescence characteristic curve is improved, and thus the stability and accuracy of the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment are improved.

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

[0059] A1: Under the fluorescence intensity under 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 under different excitation wavelengths is taken as the dependent variable, linear regression fitting is performed, and the fluorescence characteristic coefficient of the fluorescent standard substance is obtained.

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

[0061] The present scheme uses the fluorescent standard substance to replace the live algae for the calibration of the fluorescence characteristic curve of the fluorescence method algae classification equipment. The live algae has poor stability and poor batch repeatability, and is difficult to perform value traceability in the laboratory. The fluorescent substance is a standardized reagent, has strong stability, good batch repeatability, can perform value traceability, and ensures the accuracy of the fluorescence characteristic curve.

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

[0063] Figure 2 In the formula, the abscissa represents the excitation wavelength, and the unit is nanometer (nm); the ordinate represents the fluorescence characteristic coefficient.

[0064] S103: Determine the fluorescence characteristic curve 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 fluorescence standard substance and the algae fluorescence characteristic coefficient.

[0065] In S103, the fluorescence characteristic curve of different types of algae is normalized based on the characteristic curve of the fluorescent substance solution to establish the relationship model of the corresponding relationship between the fluorescence standard substance and the algae fluorescence characteristic coefficient.

[0066] Due to the influence of installation and manufacturing process of light source, lens, detector and the like, even the same type of equipment, there are still some differences between each. Therefore, the fluorescence characteristic curve of different types of algae is normalized to establish the relationship between the fluorescence standard substance and the algae fluorescence characteristic curve, and to eliminate the influence of different instrument responses on the fluorescence intensity. Also, the relationship model of the fluorescence standard substance and the algae fluorescence characteristic curve can be applied between different devices.

[0067] The process of determining the fluorescence characteristic curve of different types of algae according to the characteristic curve of the fluorescent substance solution and the pre-established relationship model is shown as B1-B3.

[0068] B1: Determine the pure algae sample by the detected chlorophyll concentration and the identified type of algae solution; wherein the pure algae at least includes green algae, blue-green algae, diatom and cryptomonad.

[0069] B2: Perform fluorescence spectrum scanning on the fixed concentration fluorescent standard substance water sample and the pure algae sample to establish the corresponding relationship between the fluorescence standard substance and the algae fluorescence characteristic coefficient in the relationship model.

[0070] Specifically, by detecting the concentration of chlorophyll a and identifying the type by morphology / molecular biology, 4 types of pure algae (green algae, blue-green algae, diatom, and cryptomonad) samples are determined, and the fixed concentration fluorescent standard substance water sample and the 4 types of pure algae samples are scanned by fluorescence spectrum to establish the corresponding relationship between the fluorescence standard substance and the algae fluorescence characteristic coefficient.

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

[0072] The fluorescence characteristic curve of different types of algae is calculated by using the obtained characteristic curve of the fluorescent substance solution and the above established relationship model of the fluorescence standard substance and the algae fluorescence characteristic curve.

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

[0074] The relationship between the fluorescence standard substance and the fluorescence characteristic coefficient of algae can be established as shown in Table 1.

[0075] Table 1

[0076]

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

[0078] The fluorescence characteristic curves of different types of algae are determined as shown in Figure 3 . Figure 3 A schematic diagram showing the relationship between the fluorescence standard substance and the fluorescence characteristic curve of algae is shown.

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

[0080] S104: The fluorescence characteristic curve calibration of the fluorescence method algae classification equipment is performed through the fluorescence characteristic curves of different types of algae.

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

[0082] The fluorescence standard substance is re-measured using the fluorescence characteristic curves of different types of algae to evaluate and calibrate the accuracy of the algae classification equipment.

[0083] The settings of the fluorescence method algae classification equipment are adjusted according to the re-measurement results to ensure that the algae classification equipment can accurately classify and quantify different algae species based on the fluorescence characteristic curve.

[0084] The calibrated equipment is verified using independent algae samples to ensure the effectiveness of the calibration and the reliability of the equipment.

[0085] The process of re-measuring the algae classification concentration of the fluorescence standard substance through the fluorescence characteristic curves of different types of algae to verify the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment is described in detail in combination with Table 2.

[0086] Table 2

[0087]

[0088] Table 2 shows the preparation of fluorescent substance solutions with concentrations of 20 μg / L and 40 μg / L. Multi-wavelength excitation light (365 nm, 410 nm, 440 nm, 470 nm, 530 nm, 570 nm, 590 nm) was sequentially incident on the water samples, and the fluorescence intensity produced by the solutions under excitation light was measured. Using the characteristic curves of the obtained fluorescent substance solutions and the established relationship between the fluorescence standard material and the algal fluorescence characteristic curves, the fluorescence characteristic curves of different algal species were calculated. The algal classification concentrations of the fluorescent standard material were then re-measured using the fluorescence characteristic curves of different algal species to verify the calibration of the fluorescence characteristic curve of the fluorescence algal classification equipment.

[0089] This method uses fluorescent standard materials instead of live algae for the calibration of fluorescence characteristic curves in fluorescence algae classification equipment. Specifically, the fluorescent material needs to have strong fluorescence characteristics at excitation wavelengths of 300-600 nm and emission wavelengths of 610-750 nm.

[0090] The fluorescence characteristic curves of different algal species are shown in Table 3.

[0091] Table 3

[0092]

[0093] To facilitate understanding the calibration process of fluorescence characteristic curves in algal classification equipment, combined with... Figure 4 For example:

[0094] Figure 4 In this process, fixed concentration fluorescent standard material water samples are identified through material certificates and concentration traceability of computer equipment;

[0095] Four pure algae (green algae, blue algae, diatoms, and cryptophytes) samples were identified by laboratory testing of chlorophyll a concentration and morphological / molecular biological identification.

[0096] Fluorescence spectroscopy was performed on water samples with fixed concentrations of fluorescent standard material and four pure algae samples to establish the relationship between the fluorescent standard material and the fluorescence characteristic coefficients of algae.

[0097] Prepare standard fluorescent substances with gradient concentrations;

[0098] Fluorescence intensity at different excitation wavelengths was detected using an algae classification analyzer.

[0099] The fluorescence characteristic coefficient of the fluorescence standard substance is calculated according to the fluorescence intensity under different excitation wavelengths; specifically, the concentration of the fluorescence standard substance prepared by the standard fluorescence substance is taken as the independent variable, and the fluorescence intensity signals under different excitation wavelengths are taken as the dependent variable, linear regression fitting is performed, and the fluorescence characteristic coefficient of the fluorescence standard substance is obtained;

[0100] The fluorescence characteristic curve of different types of algae is determined through the fluorescence characteristic coefficient of different types of algae, the characteristic curve of the fluorescence substance solution, and the corresponding relationship between the fluorescence characteristic coefficient of the fluorescence standard substance and the algae in the pre-established relationship model.

[0101] The algae classification concentration (i.e., the actual measurement value) of the fluorescence standard substance is re-measured through the fluorescence characteristic curve of different types of algae, and the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment is verified.

[0102] The present scheme uses multiple gradient concentrations of fluorescence substance solutions, different wavelengths of excitation light are sequentially irradiated on different concentrations of fluorescence substance solutions, the fluorescence intensity generated by the solution under excitation light irradiation is measured, the linear relationship between the fluorescence intensity and the fluorescence substance concentration under different excitation wavelengths is calculated, and the characteristic curve of the fluorescence substance solution is obtained; the characteristic curve of the fluorescence substance solution is obtained, and the relationship between the fluorescence characteristic coefficient of the fluorescence standard substance and the algae is used to calculate the fluorescence characteristic curve of different types of algae, and the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment is completed. Specifically, the relationship between the fluorescence characteristic coefficient of the fluorescence standard substance and the algae is established by using multiple gradient concentrations of algae solutions and fluorescence substance solutions, different wavelengths of excitation light are sequentially irradiated on different concentrations of algae solutions and fluorescence substance solutions, the fluorescence intensity generated by the solution under excitation light irradiation is measured, the linear relationship between the fluorescence intensity and the algae or fluorescence substance concentration under different excitation wavelengths is calculated, the characteristic curve of the algae solution and the fluorescence substance solution is obtained, and the relationship between the fluorescence characteristic coefficient of the fluorescence standard substance and the algae is established.

[0103] In the embodiments of the present application, the determination and calibration of the fluorescence characteristic curve are mainly carried out by using living algae at present, and there are problems such as large amount of algae solution used, large amount of measurement work, and poor stability of measurement results in the calibration process of the living algae method. The fluorescence substance is a standardized reagent, has strong stability, good batch repeatability, can be traced to the value, and ensures the accuracy of the fluorescence characteristic curve, so the fluorescence standard substance is used instead of the living algae, the concentration of the fluorescence substance prepared by the standard fluorescence substance and the fluorescence intensity signals under different excitation wavelengths are calculated to obtain the characteristic curve of the fluorescence substance solution, the fluorescence characteristic curve of different types of algae is determined according to the characteristic curve of the fluorescence substance solution and the relationship model established in advance, and the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment is carried out, so as to improve the stability and accuracy of the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment.

[0104] Based on the above embodimentsFigure 1 The application discloses a fluorescence characteristic curve calibration method of an algae classification device, and further discloses a fluorescence characteristic curve calibration device of the algae classification device. Figure 5 As shown in the figure, the fluorescence characteristic curve calibration device of the algae classification device comprises:

[0105] A measurement unit 501 is configured to measure fluorescence intensity signals of a standard fluorescent substance under different excitation wavelengths.

[0106] A calculation unit 502 is configured to calculate a characteristic curve of a fluorescent substance solution by using the fluorescence intensity signals under different excitation wavelengths and a concentration of the fluorescent substance, wherein the concentration of the fluorescent substance is prepared by using the standard fluorescent substance.

[0107] A determination unit 503 is configured to determine fluorescence characteristic curves of different kinds of algae according to the characteristic curve of the fluorescent substance solution and a pre-established relationship model, wherein the relationship model stores a corresponding relationship between the fluorescence standard substance and algae fluorescence characteristic coefficients.

[0108] A calibration unit 504 is configured to calibrate the fluorescence characteristic curve of the fluorescence method algae classification device by using the fluorescence characteristic curves of the different kinds of algae.

[0109] Further, the measurement unit 501 is specifically configured to emit excitation light to standard fluorescent substance solutions with multiple gradient concentrations to be detected by using light emitting diodes (LEDs) of different wavelengths, so that the fluorescence intensity signals under different excitation wavelengths are measured under the alternating action of high-frequency pulses of the measurement light of the LEDs of different wavelengths.

[0110] Further, the calculation unit 502 comprises:

[0111] A linear regression fitting module is configured to perform linear regression fitting by taking the concentration of the fluorescent substance prepared by using the standard fluorescent substance as an independent variable and taking the fluorescence intensity signals under different excitation wavelengths as a dependent variable, so as to obtain fluorescence characteristic coefficients of the fluorescence standard substance.

[0112] A first determination module is configured to determine the characteristic curve of the fluorescent substance solution by using the fluorescence characteristic coefficients of the fluorescence standard substance.

[0113] Further, the determination unit 503 comprises:

[0114] A second determination module is configured to determine a pure algae sample by using a detected concentration of chlorophyll and a kind of algae solution identified, wherein the pure algae at least includes green algae, blue-green algae, diatoms and cryptophytes.

[0115] The scanning module is configured to perform fluorescence spectrum scanning on the fixed-concentration fluorescence standard substance water sample and the sample of the pure algae to establish a corresponding relationship between the fluorescence standard substance and the algae fluorescence characteristic coefficient in the relationship model.

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

[0117] Further, the fluorescence characteristic curve calibration device of the algae classification equipment further comprises:

[0118] The identification unit is configured to obtain the chlorophyll concentration of the algae solution by spectrophotometric measurement, and identify the algae solution by morphology and molecular biology to obtain a plurality of algae solutions with gradient concentrations.

[0119] Further, the calibration unit 504 is specifically configured to re-measure the algae classification concentration of the fluorescence standard substance by the fluorescence characteristic curves of different types of algae to verify the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment.

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

[0121] In the embodiments of the present application, since the determination and calibration of the fluorescence characteristic curve currently mainly adopt the live algae method, there are large amounts of algae solution used, large measurement workload, and poor stability of measurement results in the calibration process of the live algae method. The fluorescence substance is a standardized reagent, has strong stability, good batch repeatability, can perform value traceability, and ensures the accuracy of the fluorescence characteristic curve. Therefore, the fluorescence standard substance is used to replace the live algae, the fluorescence substance concentration and the fluorescence intensity signal under different excitation wavelengths obtained by preparing the fluorescence substance by the standard fluorescence substance are calculated to obtain the characteristic curve of the fluorescence substance solution, and the fluorescence characteristic curves of different types of algae are determined according to the characteristic curve of the fluorescence substance solution and the relationship model established in advance to calibrate the fluorescence characteristic curve of the fluorescence method algae classification equipment, thereby improving the stability and accuracy of the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment.

[0122] The embodiments of the present application also provide a storage medium comprising stored instructions, wherein the instructions control the device where the storage medium is located to perform the fluorescence characteristic curve calibration method of the algae classification equipment as described above when the instructions are executed.

[0123] The embodiments of the present application also provide an electronic device, and a structure diagram thereof is as shown in Figure 6As shown, specifically includes a memory 601, and one or more instructions 602, wherein one or more instructions 602 stored in the memory 601, and configured to be executed by one or more processors 603 to execute the above-mentioned algal classification device fluorescence characteristic curve calibration method.

[0124] For each method embodiment described above, for the sake of simple description, it is expressed as a series of action combinations, but those skilled in the art should know that the application is not limited by the order of the described actions, because according to the application, certain steps can be performed in other order or simultaneously. Secondly, those skilled in the art should know that the embodiments described in the specification are all preferred embodiments, and the actions and modules involved are not necessarily required by the application.

[0125] It should be noted that each embodiment in the specification is described in a progressive manner, and each embodiment focuses on the difference from other embodiments. The same and similar parts between each embodiment can be referred to each other. For device embodiments, since they are basically similar to method embodiments, they are described more simply, and the relevant parts are referred to the part of the method embodiment.

[0126] The steps in the method of each embodiment of the application can be adjusted, combined and deleted according to actual needs.

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

[0128] The above description of the disclosed embodiments enables those skilled in the art to implement or use the application. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the scope of the application. Therefore, the 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 the preferred embodiment of the application. It should be noted that for those skilled in the art, without departing from the principles of the application, a number of improvements and refinements can be made, which should be considered as the protection scope of the application.

Claims

1. A method of calibrating a fluorescence characteristic curve of an algal classification device, characterized by, The method comprises: Measuring the fluorescence intensity signal of the fluorescence standard substance under different excitation wavelengths; Calculating the characteristic curve of the fluorescence substance solution through the fluorescence intensity signal under the different excitation wavelengths and the fluorescence substance concentration, wherein the fluorescence substance concentration is prepared by the fluorescence standard substance; According to the characteristic curve of the fluorescence substance solution and the pre-established relationship model, the fluorescence characteristic curve of different types of algae is determined; wherein the corresponding relationship between the fluorescence standard substance and the algae fluorescence characteristic coefficient is stored in the relationship model; Through the fluorescence characteristic curve of the different types of algae, the fluorescence characteristic curve calibration of the fluorescence method algae classification equipment is carried out; The calculating the characteristic curve of the fluorescence substance solution through the fluorescence intensity signal under the different excitation wavelengths and the fluorescence substance concentration comprises: Under the fluorescence intensity under different excitation wavelengths, the fluorescence substance concentration prepared by the fluorescence standard substance is taken as the independent variable, and the fluorescence intensity signal under different excitation wavelengths is taken as the dependent variable, linear regression fitting is carried out, and the fluorescence characteristic coefficient of the fluorescence standard substance is obtained; The fluorescence characteristic curve of the fluorescence substance solution is determined through the fluorescence characteristic coefficient of the fluorescence standard substance; The determining the fluorescence characteristic curve of different types of algae according to the characteristic curve of the fluorescence substance solution and the pre-established relationship model comprises: Through the detected chlorophyll concentration and the type obtained by identifying the algae solution, a pure algae sample is determined; wherein the pure algae at least includes green algae, blue-green algae, diatoms and cryptophytes; The fluorescence spectrum scanning is carried out on the fixed concentration fluorescence standard substance water sample and the pure algae sample, so as to establish the corresponding relationship between the fluorescence standard substance and the algae fluorescence characteristic coefficient in the relationship model; The fluorescence characteristic curve of different types of algae is determined through the characteristic curve of the fluorescence substance solution and the corresponding relationship between the fluorescence standard substance and the algae fluorescence characteristic coefficient in the established relationship model.

2. The method of claim 1, wherein, The measuring the fluorescence intensity signal of the fluorescence standard substance under different excitation wavelengths comprises: Through the light-emitting diodes of different wavelengths, excitation light is emitted to the fluorescence standard substance solution with multiple gradient concentrations to be detected, so that the fluorescence intensity signal under different excitation wavelengths is measured under the alternating action of the light-emitting diodes of different wavelengths measuring light high-frequency pulses.

3. The method of claim 1, wherein, Further comprising: The chlorophyll concentration of the algae solution is measured by spectrophotometry, and the algae solution is identified by morphology and molecular biology to obtain multiple gradient concentrations of algae solution.

4. The method of claim 1, wherein, The fluorescence characteristic curve calibration of the fluorescence method algae classification equipment is carried out through the fluorescence characteristic curve of the different types of algae. The process of establishing the relationship model comprises:

5. The method of claim 1, wherein, Taking the characteristic curve of the fluorescence substance solution as a benchmark, the fluorescence characteristic curve of different types of algae is normalized to establish the relationship model of the corresponding relationship between the fluorescence standard substance and the algae fluorescence characteristic coefficient. The device comprises:

6. A fluorescence characteristic curve calibration device of an algal classification apparatus, characterized by, A measurement unit for measuring the fluorescence intensity signal of the fluorescence standard substance under different excitation wavelengths; ​ The computing unit is configured to calculate a characteristic curve of the fluorescent substance solution by the fluorescent intensity signals at different excitation wavelengths and the concentration of the fluorescent substance, wherein the concentration of the fluorescent substance is prepared by a fluorescent standard substance. The determining unit is configured to determine the fluorescent 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 a corresponding relationship between the fluorescent standard substance and the fluorescent characteristic coefficients of algae. The calibration unit is configured to calibrate the fluorescent characteristic curves of the fluorescent algae classification equipment by the fluorescent characteristic curves of different types of algae. The computing unit comprises: The linear regression fitting module is configured to perform linear regression fitting by taking the concentration of the fluorescent substance prepared by the fluorescent standard substance as an independent variable and taking the fluorescent intensity signals at different excitation wavelengths as a dependent variable, to obtain the fluorescent characteristic coefficients of the fluorescent standard substance. The first determining module is configured to determine the characteristic curve of the fluorescent substance solution by the fluorescent characteristic coefficients of the fluorescent standard substance. The determining unit comprises: The second determining module is configured to determine the pure algae sample by the detected concentration of chlorophyll and the type of the algae solution identified, wherein the pure algae at least includes green algae, blue-green algae, diatoms and cryptophytes. The scanning module is configured to perform fluorescent spectrum scanning on the sample of the pure algae and the fluorescent standard substance water sample with a fixed concentration, to establish the corresponding relationship between the fluorescent standard substance and the fluorescent characteristic coefficients of algae in the relationship model. The third determining module is configured to determine the fluorescent characteristic curves of different types of algae by the characteristic curve of the fluorescent substance solution and the corresponding relationship between the fluorescent standard substance and the fluorescent characteristic coefficients of algae in the established relationship model.

7. The apparatus of claim 6, wherein, The measuring unit is specifically configured to: The different wavelengths of light-emitting diodes emit excitation light to the fluorescent standard substance solution with multiple gradient concentrations to be detected, so that the fluorescent intensity signals at different excitation wavelengths are measured under the alternating action of the high-frequency pulse of the light-emitting diodes at different wavelengths.

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