Fluorescence spectrum correction method and device
By determining the reference fluorescence spectrometer and correction solution and calculating the fluorescence intensity correction vector, the problem of spectral inaccuracy caused by hardware differences in the fluorescence instrument was solved, and the data quality and accuracy of the fluorescence spectrometer were improved.
Patent Information
- Application Number
- CN202211654376.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2022-12-20
- Publication Date
- 2025-09-23
- Estimated Expiration
- 2042-12-20
AI Technical Summary
Existing fluorescence correction methods cannot eliminate the intensity differences caused by the fluorescence instrument hardware, and it is difficult to improve the spectral quality in areas with low fluorescence intensity, resulting in insufficient accuracy of the correction results.
By determining the reference fluorescence spectrometer and the correction solution, the fluorescence spectra of the two are obtained, the fluorescence intensity correction vector is calculated, the correction vector of the fluorescence excitation or emission spectrum is obtained, and the spectrum correction result is obtained based on the fluorescence data and the correction vector, including the calculation of the three-dimensional fluorescence intensity correction matrix.
Effectively reduce the spectral differences of the same sample on different fluorescence spectrometers due to instrument hardware differences, improve the quality of ultraviolet fluorescence signals, and enhance data accuracy and reliability.
Smart Images
Figure CN115791737B_ABST
Abstract
Description
Technical Field
[0001] The present application relates to the field of fluorescence spectrum technology, and in particular to a fluorescence spectrum correction method and device. Background Art
[0002] Fluorescence spectroscopy technology can be used to reflect the composition of fluorescent organic matter through the specific relationship between fluorescence spectra and the specific structure of fluorescent substances. It is used in chemical analysis and detection fields such as chemistry, chemical engineering, materials, food, biology, and environment. The test is simple, fast, has no secondary pollution, and is highly sensitive.
[0003] In related technologies, due to the differences in the core components of different fluorescence spectrometers, the fluorescence signals output by the fluorescence spectrometers are different, the obtained fluorescence spectra are different, and the differences are significant in the ultraviolet light band containing rich fluorescence information, making it impossible to directly compare the fluorescence spectra obtained by different fluorescence spectrometers. Therefore, the fluorescence spectrum needs to be corrected and calculated. Common fluorescence correction methods include quinine sulfate standard solution correction, pure water Raman correction, and metal ion-doped glass standard reference correction, all of which use local fluorescence or Raman signals for spectral correction to achieve normalization of fluorescence intensity.
[0004] However, in the related technology, the existing fluorescence correction method cannot eliminate the fluorescence intensity differences caused by the instrument hardware, and it is difficult to effectively improve the fluorescence spectrum quality of some areas with low fluorescence intensity, resulting in insufficient accuracy of fluorescence spectrum correction and reducing the data quality of the obtained correction results, which urgently needs to be solved. Summary of the Invention
[0005] The present application provides a fluorescence spectrum correction method and device to solve the problems in the related art, such as the inability of existing fluorescence correction methods to eliminate the fluorescence intensity differences caused by instrument hardware, and the difficulty in effectively improving the fluorescence spectrum quality of some areas with low fluorescence intensity, resulting in insufficient accuracy of fluorescence spectrum correction and reduced data quality of the obtained correction results.
[0006] A first aspect of the present application provides a fluorescence spectrum correction method, comprising the following steps: determining a reference fluorescence spectrometer and a correction solution; obtaining the fluorescence spectra of the correction solution on the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected; calculating a fluorescence intensity correction vector based on the fluorescence spectra of the correction solution on the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected to obtain a correction vector for the fluorescence excitation or emission spectrum; and obtaining a fluorescence spectrum correction result based on the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector.
[0007] Optionally, in one embodiment of the present application, before the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector are combined to obtain the fluorescence spectrum correction result, it also includes: judging whether the fluorescence spectrum of the fluorescence spectrometer to be corrected is a three-dimensional fluorescence spectrum; if the fluorescence spectrum of the fluorescence spectrometer to be corrected is the three-dimensional fluorescence spectrum, obtaining a three-dimensional fluorescence intensity correction matrix according to the correction vector of the fluorescence excitation or emission spectrum; and obtaining the fluorescence spectrum correction result according to the fluorescence data of the fluorescence spectrometer to be corrected and the three-dimensional fluorescence intensity correction matrix.
[0008] In one embodiment of the present application, the calculation formula of the three-dimensional fluorescence intensity correction matrix is:
[0009]
[0010] Where M is the three-dimensional fluorescence correction matrix, is the excitation correction vector, is the launch correction vector.
[0011] In addition, in one embodiment of the present application, the fluorescence intensity correction vector is calculated based on the fluorescence spectra of the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected based on the correction solution to obtain the correction vector of the fluorescence excitation or emission spectrum, including: selecting the excitation or emission wavelength range within the preset correction area respectively; performing excitation correction or emission correction according to the fluorescence intensity at the corresponding wavelength to obtain the correction vector at the corresponding wavelength.
[0012] A second aspect of the present application provides a fluorescence spectrum correction device, including: a confirmation module for determining a reference fluorescence spectrometer and a correction solution; an acquisition module for obtaining the fluorescence spectra of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected; a calculation module for calculating a fluorescence intensity correction vector based on the fluorescence spectra of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected, to obtain a correction vector of the fluorescence excitation or emission spectrum; and a correction module for obtaining a fluorescence spectrum correction result based on the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector.
[0013] Optionally, in one embodiment of the present application, the correction module includes: a judgment unit, used to judge whether the fluorescence spectrum of the fluorescence spectrometer to be corrected is a three-dimensional fluorescence spectrum before the fluorescence data of the fluorescence spectrometer to be corrected is combined with the correction vector to obtain the fluorescence spectrum correction result; a first acquisition unit, used to obtain a three-dimensional fluorescence intensity correction matrix according to the correction vector of the fluorescence excitation or emission spectrum when the fluorescence spectrum of the fluorescence spectrometer to be corrected is the three-dimensional fluorescence spectrum; and a generation unit, used to obtain the fluorescence spectrum correction result according to the fluorescence data of the fluorescence spectrometer to be corrected and the three-dimensional fluorescence intensity correction matrix.
[0014] In one embodiment of the present application, the calculation formula of the three-dimensional fluorescence intensity correction matrix is:
[0015]
[0016] Where M is the three-dimensional fluorescence correction matrix, is the excitation correction vector, is the launch correction vector.
[0017] In addition, in one embodiment of the present application, the calculation module includes: a selection unit for selecting the excitation or emission wavelength range within the preset correction area respectively; a second acquisition unit for performing excitation correction or emission correction according to the fluorescence intensity at the corresponding wavelength to obtain the correction vector at the corresponding wavelength.
[0018] The third aspect of the present application provides an electronic device, comprising: a memory, a processor, and a computer program stored in the memory and executable on the processor, wherein the processor executes the program to implement the fluorescence spectrum correction method as described in the above embodiment.
[0019] A fourth aspect of the present application provides a computer-readable storage medium, which stores a computer program that implements the above fluorescence spectrum correction method when executed by a processor.
[0020] The embodiment of the present application can determine the reference fluorescence spectrometer and the correction solution, obtain the fluorescence spectra of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected, calculate the fluorescence intensity correction vector based on the fluorescence spectra of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected, obtain the correction vector of the fluorescence excitation or emission spectrum, and obtain the fluorescence spectrum correction result based on the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector. It can effectively reduce the spectral differences of the same sample on different fluorescence spectrometers due to instrument hardware differences, while improving the fluorescence signal quality of the sample in the ultraviolet band, enhancing the data quality of the fluorescence spectrometer, and making it more accurate and reliable. Therefore, it solves the problems in the related art that the existing fluorescence correction method cannot eliminate the fluorescence intensity differences caused by instrument hardware, and it is difficult to effectively improve the fluorescence spectrum quality of some areas with low fluorescence intensity, resulting in insufficient accuracy of fluorescence spectrum correction and reduced data quality of the obtained correction results.
[0021] Additional aspects and advantages of the present application will be given in part in the description below, and in part will become apparent from the description below, or will be learned through practice of the present application. BRIEF DESCRIPTION OF THE DRAWINGS
[0022] The above and / or additional aspects and advantages of the present application will become apparent and easily understood from the following description of the embodiments in conjunction with the accompanying drawings, in which:
[0023] Figure 1 A flowchart of a fluorescence spectrum correction method provided according to an embodiment of the present application;
[0024] Figure 2 A three-dimensional fluorescence spectrum of a sample measured by fluorescence instrument B according to an embodiment of the present application;
[0025] Figure 3 The excitation spectrum of quinine sulfate solution under 385 nm emission light and the correction coefficient in the excitation correction vector of one embodiment of the present application;
[0026] Figure 4 The excitation spectrum of a 4-amino-3-hydroxy-1-naphthalenesulfonic acid solution under 445 nm emission light and the correction coefficient in the excitation correction vector of an embodiment of the present application;
[0027] Figure 5 The excitation spectrum of a Rhodamine B solution under 575 nm emission light and the correction coefficient in the excitation correction vector of an embodiment of the present application;
[0028] Figure 6 The emission spectrum of the mixed solution under 235 nm excitation light and the correction coefficient in the emission correction vector of an embodiment of the present application;
[0029] Figure 7 The emission spectrum of the mixed solution under 345 nm excitation light and the correction coefficient in the emission correction vector of an embodiment of the present application;
[0030] Figure 8 The correction coefficients in the excitation and emission correction vectors when the fluorometer B corrects the fluorometer A in one embodiment of the present application;
[0031] Figure 9 This is a contour map of a three-dimensional fluorescence spectrum correction matrix when correction is performed from fluorometer B to fluorometer A according to one embodiment of the present application;
[0032] Figure 10 A sample of an embodiment of this application Figure 9 Three-dimensional fluorescence spectrum after correction by the correction matrix;
[0033] Figure 11 Schematic diagram of the structure of a fluorescence spectrum correction device provided according to an embodiment of the present application;
[0034] Figure 12 A schematic diagram of the structure of an electronic device provided according to an embodiment of the present application. DETAILED DESCRIPTION
[0035] The following describes in detail embodiments of the present application, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present application, and should not be construed as limiting the present application.
[0036] The following describes the fluorescence spectrum correction method and device of the embodiment of the present application with reference to the accompanying drawings. In view of the related technologies mentioned in the above background technology center, the existing fluorescence correction method cannot eliminate the fluorescence intensity difference caused by the instrument hardware, and it is difficult to effectively improve the fluorescence spectrum quality of some areas with lower fluorescence intensity, which makes the accuracy of fluorescence spectrum correction insufficient and reduces the data quality of the obtained correction result. The present application provides a fluorescence spectrum correction method, which determines a reference fluorescence spectrometer and a correction solution, obtains the fluorescence spectrum of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected, calculates the fluorescence intensity correction vector based on the fluorescence spectrum of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected, obtains the correction vector of the fluorescence excitation or emission spectrum, and obtains the fluorescence spectrum correction result according to the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector. It can effectively reduce the spectral difference of the same sample on different fluorescence spectrometers caused by the difference in instrument hardware, and at the same time improve the fluorescence signal quality of the sample in the ultraviolet band, enhance the data quality of the fluorescence spectrometer, and make it more accurate and reliable. This solves the problems in the related art, such as the inability of existing fluorescence correction methods to eliminate the fluorescence intensity differences caused by instrument hardware, and the difficulty in effectively improving the fluorescence spectrum quality in some areas with low fluorescence intensity, resulting in insufficient accuracy of fluorescence spectrum correction and reduced data quality of the obtained correction results.
[0037] Specifically, Figure 1 A schematic flow chart of a fluorescence spectrum correction method provided in an embodiment of the present application.
[0038] like Figure 1 As shown, the fluorescence spectrum correction method includes the following steps:
[0039] In step S101 , a reference fluorescence spectrometer and a correction solution are determined.
[0040] It can be understood that the reference fluorescence spectrometer in the embodiment of the present application can be a standard instrument relative to the fluorescence spectrometer to be corrected, and a solution with an appropriate concentration of a fluorescent compound can be selected as a correction solution.
[0041] In the actual implementation process, when selecting fluorescent compounds, the principles of simplicity and harmlessness, interval coverage, minimum quantity, mixing stability and appropriate concentration must be met. Among them, the principle of simplicity and harmlessness requires that the fluorescent compound should be easy to obtain, and the correction solution configured with it should be stable and not easy to deteriorate. The interval coverage principle requires that the excitation and emission wavelength ranges of the correction solution should cover the wavelength range of the fluorescence data to be corrected. The principle of minimum quantity requires that when one fluorescent compound is not enough to cover the wavelength range, a combination of fluorescent compounds with as few as possible quantities should be selected to achieve the interval coverage principle, and the edges of the fluorescence wavelength ranges of each fluorescent compound should overlap with each other. The principle of mixing stability requires that when the situation of the minimum quantity principle occurs, it should be ensured that the mixed solution of the selected fluorescent compounds does not react in a short period of time and does not affect the fluorescence stability of the mixed solution. The principle of appropriate concentration requires that the concentration of the configured correction solution should be sufficient to measure obvious fluorescence signals on the fluorescence spectrometer to be tested.
[0042] The embodiment of the present application can determine a reference fluorescence spectrometer and a correction solution to provide the required correction data for the fluorescence spectrometer to be corrected, thereby further realizing the fluorescence spectrum correction in the following steps.
[0043] In step S102 , the fluorescence spectra of the correction solution on the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected are obtained.
[0044] It can be understood that when obtaining the fluorescence spectra of the correction solution on different fluorescence spectrometers in the embodiment of the present application, the fluorescence wavelength range, step wavelength and other test conditions of the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected are set to be consistent during the test process, and the fluorescence intensity, wavelength range, step wavelength and other information of the fluorescence spectrum output are saved during the test.
[0045] For example, for fluorescence spectrometers A and B, where fluorescence spectrometer A is the reference fluorescence spectrometer and fluorescence spectrometer B is the fluorescence spectrometer to be corrected, there are n correction solutions for excitation correction and m correction solutions for emission correction, then the corresponding excitation correction wavelength ranges under n emission wavelengths and the corresponding emission correction wavelength ranges under m excitation wavelengths are obtained.
[0046] The embodiment of the present application can obtain the fluorescence spectra of the correction solution on the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected. By using several correction solutions to perform tests on different fluorescence spectrometers, the coverage of the correction wavelength range is expanded.
[0047] In step S103 , a fluorescence intensity correction vector is calculated based on the fluorescence spectra of the correction solution on the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected, to obtain a correction vector of the fluorescence excitation or emission spectrum.
[0048] It can be understood that in the process of calculating the fluorescence intensity correction vector in the embodiment of the present application, the excitation correction spectra of the obtained correction solution can be arranged in order from small to large wavelength, and the emission correction spectra can also be arranged in order from small to large wavelength, so that the excitation or emission wavelength ranges of the correction solution are connected to each other, and then the correction vector at the corresponding wavelength is calculated and obtained.
[0049] In the actual implementation process, let EX (excitation) represent excitation, EM (emission) represent emission, and the calculation formulas of excitation correction vector and emission correction vector are respectively
[0050]
[0051]
[0052] in, is the excitation correction vector, is the emission correction vector, To express the concatenation operation of the two vectors, the correction vector formula is stimulated by
[0053]
[0054] It means the ith excitation correction solution at the emission wavelength EM i Corrected excitation wavelength range under The ratio of the fluorescence excitation vector of instrument A to the corresponding element of instrument B still forms a vector, and the emission correction vector formula is the same. i,1 is the starting wavelength of the excitation correction wavelength range corresponding to the correction solution i, in nm, EX i,ti is the end wavelength of the excitation correction wavelength range corresponding to the correction solution i, in nm, EM j,1 is the starting wavelength of the emission correction wavelength range corresponding to the correction solution j, in nm, EM j,uj is the end wavelength of the emission correction wavelength range corresponding to the correction solution j, in nm. is the excitation correction vector connection coefficient corresponding to the correction solution i, θ j is the emission correction vector connection coefficient corresponding to the correction solution j, both are dimensionless values, and their calculation formulas are
[0055]
[0056]
[0057] in, It means that the first excitation correction solution emits at wavelength EM iThe ratio of the fluorescence intensities obtained by instruments A and B at the kth excitation wavelength within the excitation correction wavelength range under Similarly, they are all dimensionless values. And when i=1, When j = 1, θ j =1.
[0058] The embodiment of the present application can calculate the fluorescence intensity correction vector based on the fluorescence spectra of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected, and obtain the correction vector of the fluorescence excitation or emission spectrum, thereby providing the required information for implementing the data correction in the following steps, so as to further realize the spectral correction of the fluorescence spectrometer.
[0059] In one embodiment of the present application, the calculation formula of the three-dimensional fluorescence intensity correction matrix is:
[0060]
[0061] Where M is the three-dimensional fluorescence correction matrix, is the excitation correction vector, is the launch correction vector.
[0062] As can be seen from the above formula, the embodiment of the present application can calculate the correction vectors of the fluorescence excitation and emission spectra in the above steps to obtain a three-dimensional fluorescence intensity correction matrix of the three-dimensional fluorescence spectrum.
[0063] In addition, in one embodiment of the present application, a fluorescence intensity correction vector is calculated based on the fluorescence spectra of the correction solution in a reference fluorescence spectrometer and a fluorescence spectrometer to be corrected to obtain a correction vector for the fluorescence excitation or emission spectrum, including: selecting an excitation or emission wavelength range within a preset correction area; performing excitation correction or emission correction according to the fluorescence intensity at the corresponding wavelength to obtain a correction vector at the corresponding wavelength.
[0064] It can be understood that the preset correction area selected in the embodiment of the present application can be a certain specific wavelength range, and the excitation or emission wavelength range within the correction area is selected within this range. The corresponding fluorescence intensity is obtained according to the selected wavelength range, and then the excitation correction or emission correction is performed according to the fluorescence intensity at the corresponding wavelength to obtain the correction vector at the corresponding wavelength.
[0065] It should be noted that the preset correction area is set by those skilled in the art according to actual conditions and is not specifically limited here.
[0066] In the embodiment of the present application, the excitation or emission wavelength range within the preset correction area can be selected respectively, and the excitation correction or emission correction can be performed according to the fluorescence intensity at the corresponding wavelength to obtain the correction vector at the corresponding wavelength. Then, the data of the wavelength intervals obtained from the several correction solutions in the above steps can be utilized to obtain the correction vectors at the corresponding excitation or emission wavelengths respectively.
[0067] In step S104 , a fluorescence spectrum correction result is obtained according to the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector.
[0068] It can be understood that in the embodiment of the present application, the fluorescence data of the fluorescence spectrometer to be corrected can be the intensity vector of the fluorescence excitation and emission spectra measured by the fluorescence spectrometer to be corrected, and the corresponding three-dimensional fluorescence spectrum matrix, which is calculated by the reference fluorescence spectrometer and the correction vector. The result obtained is the correction result of the fluorescence spectrometer to be corrected, which can be directly compared with the reference fluorescence spectrometer.
[0069] The embodiment of the present application can obtain a fluorescence spectrum correction result based on the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector, thereby realizing the comparison between the data obtained by different fluorescence spectrometers, improving the data quality of the obtained data, simplifying the process of data comparison between different instruments, and making it more accurate.
[0070] Optionally, in one embodiment of the present application, before the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector are combined to obtain the fluorescence spectrum correction result, it also includes: judging whether the fluorescence spectrum of the fluorescence spectrometer to be corrected is a three-dimensional fluorescence spectrum; if the fluorescence spectrum of the fluorescence spectrometer to be corrected is a three-dimensional fluorescence spectrum, obtaining a three-dimensional fluorescence intensity correction matrix according to the correction vector of the fluorescence excitation or emission spectrum; and obtaining the fluorescence spectrum correction result according to the fluorescence data of the fluorescence spectrometer to be corrected and the three-dimensional fluorescence intensity correction matrix.
[0071] It can be understood that the three-dimensional fluorescence intensity correction matrix in the embodiment of the present application can be calculated and obtained by the calculation formula of the three-dimensional fluorescence intensity correction matrix in the above steps, and then calculated by the fluorescence data, correction vector and three-dimensional fluorescence intensity correction matrix of the fluorescence spectrometer to be corrected to obtain the fluorescence spectrum correction result of the fluorescence spectrometer to be corrected.
[0072] In the actual implementation process, still refer to the above steps, set the reference fluorescence spectrometer as fluorescence spectrometer A, the fluorescence spectrometer to be corrected as fluorescence spectrometer B, and the intensity vector of the fluorescence excitation spectrum, the intensity vector of the emission spectrum and the three-dimensional fluorescence spectrum matrix to be corrected for fluorescence spectrometer A are respectively
[0073]
[0074]
[0075] EEM B→A =EM B ⊙M,
[0076] in, is the intensity vector of the fluorescence excitation spectrum corrected for fluorescence spectrometer A, is the intensity vector of the fluorescence excitation spectrum measured by fluorescence spectrometer B, is the excitation correction vector, is the emission spectrum intensity vector for correction to fluorescence spectrometer A, is the intensity vector of the fluorescence emission spectrum measured by fluorescence spectrometer B, EEM is the emission correction vector B→A To correct the three-dimensional fluorescence spectrum matrix for fluorescence spectrometer A, EEM B is the three-dimensional fluorescence spectrum matrix of fluorescence spectrometer B, M is the three-dimensional fluorescence correction matrix, and ⊙ represents the Hadamard product operation of two vectors or matrices. and EEM B→A , complete the correction processing based on fluorescence spectrometer A, and then directly compare it with the data of fluorescence spectrometer A.
[0077] The embodiment of the present application can determine whether the fluorescence spectrum of the fluorescence spectrometer to be corrected is a three-dimensional fluorescence spectrum. If the fluorescence spectrum of the fluorescence spectrometer to be corrected is a three-dimensional fluorescence spectrum, a three-dimensional fluorescence intensity correction matrix is obtained according to the correction vector of the fluorescence excitation or emission spectrum, and a fluorescence spectrum correction result is obtained according to the fluorescence data of the fluorescence spectrometer to be corrected and the three-dimensional fluorescence intensity correction matrix. By calculating and processing the obtained three-dimensional fluorescence spectrum data of the fluorescence spectrometer, correction and comparison of the three-dimensional fluorescence spectrum data are realized, making the fluorescence spectrum correction more practical.
[0078] The following combination Figure 2-10 , the working content of the embodiment of this application is described in detail with a specific embodiment.
[0079] For fluorescence spectrometers A and B, the data quality of fluorescence spectrometer A is higher, while Figure 2 As shown, the data quality of fluorescence spectrometer B in the ultraviolet region is poor. At this time, the measurement parameters of the three-dimensional fluorescence spectrum are set as an excitation wavelength range of 220-550nm, an emission wavelength range of 250-600nm, and a step wavelength of 5nm.
[0080] According to the correction substance selection criteria, excitation spectrum correction from 220 nm to 550 nm was performed using 0.1 mg / L quinine sulfate aqueous solution, 0.02 mg / L 4-amino-3-hydroxy-1-naphthalenesulfonic acid aqueous solution, and 0.1 mg / L rhodamine B aqueous solution. For excitation spectrum correction, 0.1 mg / L quinine sulfate aqueous solution was used to correct the excitation light from 220 nm to 315 nm, transitioning from 275 nm to the signal from 0.02 mg / L 4-amino-3-hydroxy-1-naphthalenesulfonic acid aqueous solution. 0.02 mg / L 4-amino-3-hydroxy-1-naphthalenesulfonic acid aqueous solution was used to correct the excitation light from 235 nm to 365 nm, transitioning from 355 nm to the signal from 0.1 mg / L rhodamine B aqueous solution. 0.1 mg / L rhodamine B aqueous solution was used to correct the excitation light from 310 nm to 550 nm. The emission spectrum at 250nm-600nm was corrected using a mixed aqueous solution of tryptophan, tyrosine, quinine sulfate, 4-amino-3-hydroxy-1-naphthalenesulfonic acid and rhodamine B.
[0081] The fluorescence spectrum of fluorescence spectrometer B is corrected using fluorescence spectrometer A as a benchmark.
[0082] As shown in Table 1, the wavelength range of the fluorescence correction solution is used for correction. According to Table 1, the excitation or emission spectrum of each fluorescence correction solution is obtained, and the fluorescence correction solution is obtained by referring to Table 1. Figure 3-7 , respectively obtain the relationship between each element and wavelength in the correction vector of each fluorescence correction solution in each wavelength range, and then refer to Figure 8 , connecting each segment of excitation or emission correction vector into the overall excitation and emission correction coefficient vector.
[0083] Table 1
[0084]
[0085] Then calculate the three-dimensional fluorescence correction coefficient matrix, such as Figure 9 As shown in , the coefficients in the correction matrix corresponding to each excitation and emission wavelength coordinate are represented in the form of a contour map.
[0086] Figure 2 is the three-dimensional fluorescence spectrum of fluorescence spectrometer B before correction. After correction calculation, the three-dimensional fluorescence spectrum of a sample tested by fluorescence spectrometer B can be obtained as follows: Figure 10 The three-dimensional fluorescence spectrum after correction based on fluorescence spectrometer A is shown. It can be seen that in the ultraviolet region with a wavelength less than 380nm, the fluorescence data has achieved a good correction effect.
[0087] According to the fluorescence spectrum correction method proposed in the embodiment of the present application, the fluorescence spectrum of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected can be obtained by determining the reference fluorescence spectrometer and the correction solution, and the fluorescence intensity correction vector is calculated based on the fluorescence spectrum of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected to obtain the correction vector of the fluorescence excitation or emission spectrum. The fluorescence spectrum correction result is obtained based on the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector. This can effectively reduce the spectral difference caused by instrument hardware differences of the same sample on different fluorescence spectrometers, while improving the fluorescence signal quality of the sample in the ultraviolet band, enhancing the data quality of the fluorescence spectrometer, and making it more accurate and reliable. Thus, the problem in the related art that the existing fluorescence correction method cannot eliminate the fluorescence intensity difference caused by instrument hardware, and it is difficult to effectively improve the fluorescence spectrum quality of some areas with low fluorescence intensity, resulting in insufficient accuracy of fluorescence spectrum correction and reduced data quality of the obtained correction result is solved.
[0088] Next, the fluorescence spectrum correction device proposed according to the embodiment of the present application is described with reference to the accompanying drawings.
[0089] Figure 11 Schematic diagram of a fluorescence spectrum correction device according to an embodiment of the present application.
[0090] like Figure 11 As shown, the fluorescence spectrum correction device 10 includes: a confirmation module 100 , an acquisition module 200 , a calculation module 300 and a correction module 400 .
[0091] The confirmation module 100 is used to determine the reference fluorescence spectrometer and the correction solution.
[0092] The acquisition module 200 is used to acquire the fluorescence spectra of the correction solution on the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected.
[0093] The calculation module 300 is used to calculate the fluorescence intensity correction vector based on the fluorescence spectra of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected, so as to obtain the correction vector of the fluorescence excitation or emission spectrum.
[0094] The correction module 400 is used to obtain a fluorescence spectrum correction result according to the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector.
[0095] Optionally, in one embodiment of the present application, the correction module 400 includes: a judgment unit, a first acquisition unit, and a generation unit.
[0096] The judging unit is used to judge whether the fluorescence spectrum of the fluorescence spectrometer to be corrected is a three-dimensional fluorescence spectrum before obtaining a fluorescence spectrum correction result by combining the fluorescence data of the fluorescence spectrometer to be corrected with the correction vector.
[0097] The first acquisition unit is configured to obtain a three-dimensional fluorescence intensity correction matrix according to a correction vector of the fluorescence excitation or emission spectrum when the fluorescence spectrum of the fluorescence spectrometer to be corrected is a three-dimensional fluorescence spectrum.
[0098] The generating unit is used to obtain the fluorescence spectrum correction result according to the fluorescence data of the fluorescence spectrometer to be corrected and the three-dimensional fluorescence intensity correction matrix.
[0099] In one embodiment of the present application, the calculation formula of the three-dimensional fluorescence intensity correction matrix is:
[0100]
[0101] Where M is the three-dimensional fluorescence correction matrix, is the excitation correction vector, is the launch correction vector.
[0102] In addition, in one embodiment of the present application, the calculation module 300 includes: a selection unit and a second acquisition unit.
[0103] The selection unit is used to select the excitation or emission wavelength range within the preset correction area.
[0104] The second acquisition unit is used to perform excitation correction or emission correction according to the fluorescence intensity at the corresponding wavelength to obtain a correction vector at the corresponding wavelength.
[0105] It should be noted that the above explanation of the embodiment of the fluorescence spectrum correction method is also applicable to the fluorescence spectrum correction device of this embodiment, and will not be repeated here.
[0106] According to the fluorescence spectrum correction device proposed in the embodiment of the present application, the fluorescence spectrum of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected can be obtained by determining the reference fluorescence spectrometer and the correction solution, and the fluorescence intensity correction vector is calculated based on the fluorescence spectrum of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected to obtain the correction vector of the fluorescence excitation or emission spectrum, and the fluorescence spectrum correction result is obtained according to the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector. It can effectively reduce the spectral difference caused by the instrument hardware difference of the same sample on different fluorescence spectrometers, and at the same time improve the fluorescence signal quality of the sample in the ultraviolet band, enhance the data quality of the fluorescence spectrometer, and make it more accurate and reliable. Therefore, it solves the problems in the related art that the existing fluorescence correction method cannot eliminate the fluorescence intensity difference caused by the instrument hardware, and it is difficult to effectively improve the fluorescence spectrum quality of some areas with low fluorescence intensity, resulting in insufficient accuracy of fluorescence spectrum correction and reduced data quality of the obtained correction result.
[0107] Figure 12 This is a schematic diagram of the structure of an electronic device provided in an embodiment of the present application. The electronic device may include:
[0108] A memory 1201 , a processor 1202 , and a computer program stored in the memory 1201 and executable on the processor 1202 .
[0109] When the processor 1202 executes the program, the fluorescence spectrum correction method provided in the above embodiment is implemented.
[0110] Furthermore, the electronic device further includes:
[0111] The communication interface 1203 is used for communication between the memory 1201 and the processor 1202 .
[0112] The memory 1201 is used to store computer programs that can be run on the processor 1202 .
[0113] The memory 1201 may include a high-speed RAM memory, and may also include a non-volatile memory (non-volatile memory), such as at least one disk memory.
[0114] If the memory 1201, processor 1202, and communication interface 1203 are implemented independently, the communication interface 1203, memory 1201, and processor 1202 can be connected to each other via a bus and communicate with each other. The bus can be an Industry Standard Architecture (ISA) bus, a Peripheral Component Interconnect (PCI) bus, or an Extended Industry Standard Architecture (EISA) bus. Buses can be divided into address buses, data buses, control buses, etc. For ease of representation, Figure 12 Only one thick line is used in the diagram, but this does not mean that there is only one bus or one type of bus.
[0115] Optionally, in a specific implementation, if the memory 1201, the processor 1202 and the communication interface 1203 are integrated on a chip, the memory 1201, the processor 1202 and the communication interface 1203 can communicate with each other through an internal interface.
[0116] The processor 1202 may be a central processing unit (CPU), an application specific integrated circuit (ASIC), or one or more integrated circuits configured to implement the embodiments of the present application.
[0117] This embodiment further provides a computer-readable storage medium having a computer program stored thereon. When the program is executed by a processor, the above fluorescence spectrum correction method is implemented.
[0118] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or N embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.
[0119] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be understood to indicate or imply relative importance or implicitly specify the number of technical features indicated. Thus, a feature specified as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of this application, "N" means at least two, for example, two, three, etc., unless otherwise specifically defined.
[0120] Any process or method description in a flowchart or otherwise described herein may be understood to represent a module, fragment or portion of code comprising one or N executable instructions for implementing a custom logical function or process step, and the scope of the preferred embodiments of the present application includes alternative implementations in which functions may be performed in a different order than shown or discussed, including performing functions in a substantially simultaneous manner or in a reverse order depending on the functions involved, which should be understood by those skilled in the art to which the embodiments of the present application pertain.
[0121] The logic and / or steps represented in the flowcharts or otherwise described herein, for example, can be considered as a sequenced list of executable instructions for implementing the logical functions, and can be embodied in any computer-readable medium for use by, or in conjunction with, an instruction execution system, apparatus, or device (e.g., a computer-based system, a system including a processor, or other system that can fetch and execute instructions from an instruction execution system, apparatus, or device). For purposes of this specification, a "computer-readable medium" can be any device that can contain, store, communicate, propagate, or transport a program for use by, or in conjunction with, an instruction execution system, apparatus, or device. More specific examples (a non-exhaustive list) of computer-readable media include the following: an electrical connection with one or N wires (electronic devices), a portable computer disk cartridge (magnetic device), random access memory (RAM), read-only memory (ROM), erasable and programmable read-only memory (EPROM or flash memory), fiber optic devices, and a portable compact disc read-only memory (CDROM). In addition, the computer-readable medium may even be paper or other suitable medium on which the program is printed, since the program can be obtained electronically by optically scanning the paper or other medium and then editing, interpreting or processing it in other suitable ways as necessary, and then storing it in a computer memory.
[0122] It should be understood that various parts of the present application can be implemented using hardware, software, firmware, or a combination thereof. In the above embodiment, the N steps or methods can be implemented using software or firmware stored in a memory and executed by a suitable instruction execution system. For example, if implemented using hardware, as in another embodiment, any one of the following technologies known in the art or a combination thereof can be used to implement: a discrete logic circuit having a logic gate circuit for implementing a logic function on a data signal, an application-specific integrated circuit having a suitable combination of logic gate circuits, a programmable gate array (PGA), a field programmable gate array (FPGA), etc.
[0123] Those skilled in the art will understand that all or part of the steps in the method of the above embodiment can be completed by instructing related hardware through a program, and the program can be stored in a computer-readable storage medium. When the program is executed, it includes one or a combination of the steps of the method embodiment.
[0124] In addition, the functional units in the various embodiments of the present application may be integrated into a processing module, or each unit may exist physically separately, or two or more units may be integrated into a module. The above-mentioned integrated module may be implemented in the form of hardware or in the form of a software functional module. If the integrated module is implemented in the form of a software functional module and sold or used as an independent product, it may also be stored in a computer-readable storage medium.
[0125] The storage medium mentioned above may be a read-only memory, a magnetic disk, or an optical disk, etc. Although the embodiments of the present application have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present application. Persons skilled in the art may make changes, modifications, substitutions, and variations to the above embodiments within the scope of the present application.
Claims
1. A fluorescence spectrum correction method, characterized in that: The following steps are involved: Determine the reference fluorescence spectrometer and correction solution; Obtaining fluorescence spectra of the correction solution on the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected; Calculating a fluorescence intensity correction vector based on the fluorescence spectra of the correction solution on the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected to obtain a correction vector for the fluorescence excitation or emission spectrum; as well as Obtaining a fluorescence spectrum correction result according to the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector; The step of obtaining the fluorescence spectrum correction result based on the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector includes: determining whether the fluorescence spectrum of the fluorescence spectrometer to be corrected is a three-dimensional fluorescence spectrum; If the fluorescence spectrum of the fluorescence spectrometer to be corrected is the three-dimensional fluorescence spectrum, obtaining a three-dimensional fluorescence intensity correction matrix according to the correction vector of the fluorescence excitation or emission spectrum; Obtaining the fluorescence spectrum correction result according to the fluorescence data of the fluorescence spectrometer to be corrected and the three-dimensional fluorescence intensity correction matrix; The calculation formula of the three-dimensional fluorescence intensity correction matrix is: Where M is the three-dimensional fluorescence correction matrix, is the excitation correction vector, is the launch correction vector; The calculation formulas for the excitation correction vector and the emission correction vector are: in, is the excitation correction vector, is the emission correction vector, To represent the concatenation operation of the two vectors, EX i,1 is the starting wavelength of the excitation correction wavelength range corresponding to the correction solution i, in nm, is the end wavelength of the excitation correction wavelength range corresponding to the correction solution i, in nm, EM j,1 is the starting wavelength of the emission correction wavelength range corresponding to the correction solution j, in nm, is the end wavelength of the emission correction wavelength range corresponding to the correction solution j, in nm, is the excitation correction vector connection coefficient corresponding to the correction solution i, θ j is the emission correction vector connection coefficient corresponding to the correction solution j, both are dimensionless values, For the i-th excitation correction solution at emission wavelength EM i Corrected excitation wavelength range under The ratio of the fluorescence excitation vector of instrument A to the corresponding element of instrument B.
2. The method according to claim 1, characterized in that The calculation of the fluorescence intensity correction vector based on the fluorescence spectra of the correction solution in the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected to obtain the correction vector of the fluorescence excitation or emission spectrum includes: Select the excitation or emission wavelength range within the preset correction area respectively; An excitation correction or an emission correction is performed according to the fluorescence intensity at the corresponding wavelength to obtain a correction vector at the corresponding wavelength.
3. A fluorescence spectrum correction device, characterized in that: include: a confirmation module for determining a reference fluorescence spectrometer and a correction solution; An acquisition module, configured to acquire the fluorescence spectra of the correction solution on the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected; A calculation module, configured to calculate a fluorescence intensity correction vector based on the fluorescence spectra of the correction solution on the reference fluorescence spectrometer and the fluorescence spectrometer to be corrected, to obtain a correction vector for the fluorescence excitation or emission spectrum; as well as A correction module, configured to obtain a fluorescence spectrum correction result based on the fluorescence data of the fluorescence spectrometer to be corrected and the correction vector; Wherein, the correction module includes: a judging unit, configured to judge whether the fluorescence spectrum of the fluorescence spectrometer to be corrected is a three-dimensional fluorescence spectrum before obtaining the fluorescence spectrum correction result by combining the fluorescence data of the fluorescence spectrometer to be corrected with the correction vector; a first acquiring unit, configured to obtain a three-dimensional fluorescence intensity correction matrix according to a correction vector of the fluorescence excitation or emission spectrum when the fluorescence spectrum of the fluorescence spectrometer to be corrected is the three-dimensional fluorescence spectrum; a generating unit, configured to obtain the fluorescence spectrum correction result according to the fluorescence data of the fluorescence spectrometer to be corrected and the three-dimensional fluorescence intensity correction matrix; The calculation formula of the three-dimensional fluorescence intensity correction matrix is: Where M is the three-dimensional fluorescence correction matrix, is the excitation correction vector, is the launch correction vector; The calculation formulas for the excitation correction vector and the emission correction vector are: in, is the excitation correction vector, is the emission correction vector, To represent the concatenation operation of the two vectors, EX i,1 is the starting wavelength of the excitation correction wavelength range corresponding to the correction solution i, in nm, is the end wavelength of the excitation correction wavelength range corresponding to the correction solution i, in nm, EM j,1 is the starting wavelength of the emission correction wavelength range corresponding to the correction solution j, in nm, is the end wavelength of the emission correction wavelength range corresponding to the correction solution j, in nm, is the excitation correction vector connection coefficient corresponding to the correction solution i, θ j is the emission correction vector connection coefficient corresponding to the correction solution j, both are dimensionless values, For the i-th excitation correction solution at emission wavelength EM i Corrected excitation wavelength range under The ratio of the fluorescence excitation vector of instrument A to the corresponding element of instrument B.
4. The device according to claim 3, characterized in that The calculation module includes: A selection unit, for selecting an excitation or emission wavelength range within a preset correction area; The second acquisition unit is used to perform excitation correction or emission correction according to the fluorescence intensity at the corresponding wavelength to obtain a correction vector at the corresponding wavelength.
5. An electronic device, characterized in that: include: A memory, a processor, and a computer program stored in the memory and executable on the processor, wherein the processor executes the program to implement the fluorescence spectrum correction method according to any one of claims 1 to 2.
6. A computer-readable storage medium having a computer program stored thereon, characterized in that: The program is executed by a processor to implement the fluorescence spectrum correction method according to any one of claims 1 to 2.