A method and system for quantifying the luminescence brightness of fluorescent probes with detuned excitation wavelengths

Abstract

This invention relates to the field of fluorescent probe technology, and provides a method and system for quantifying the luminescence brightness of fluorescent probes with detuned excitation wavelengths. The method for quantifying the luminescence brightness of fluorescent probes with detuned excitation wavelengths includes: constructing an initial structure of the probe molecule and optimizing the geometry of the probe molecule's ground state to obtain a stable configuration of the probe molecule's ground state; based on the stable configuration of the probe molecule's ground state, calculating the excited state and obtaining the minimum excitation intensity of the probe molecule. n The excitation energy of each excited state and its... X , Y , Z Absorption transition dipole moments in three directions; based on the stable configuration of the probe molecule's ground state, the geometry of the probe molecule's lowest excited state is optimized to obtain the emission energy of the probe molecule's lowest excited state and its... X , Y , Z The emission transition dipole moments in three directions are calculated; combined with the energy parameters and transition dipole moment parameters, the luminescence index is calculated to select the optimal probe. This achieves accurate and rapid prediction of probe luminescence.

Description

Technical Field

[0001] This invention relates to the field of fluorescent probe technology, and in particular to a method and system for quantifying the luminescence brightness of a fluorescent probe with detuned excitation wavelength. Background Technology

[0002] The statements in this section are merely background information related to the present invention and do not necessarily constitute prior art.

[0003] Fluorescent probes, as functional compounds, can emit specific fluorescent signals by binding specifically to biomolecules, thereby enabling the detection, imaging, and quantitative analysis of biomolecules. In recent years, fluorescent probes have become indispensable tools in biomedical research due to their significant advantages such as high sensitivity, high selectivity, non-destructive nature, and real-time response. They are widely used in fields such as intracellular biomolecule tracking, cell structure imaging, early disease diagnosis, and high-throughput drug screening.

[0004] Luminescence brightness, as a core indicator of the overall performance of a fluorescent probe, directly determines imaging contrast, signal-to-noise ratio, and detection limit. According to the definition of luminescence brightness, light absorption intensity and radiative transition rate are key factors affecting it. Typically, scanning absorption spectra is used in experiments to determine the absorption peak wavelength; excitation light at this wavelength will produce the probe with the highest brightness. However, this method has the following limitations: first, it relies on expensive wavelength-tunable lasers; second, the testing cycle is long, and material and labor costs are high; third, experimental results are easily affected by differences in sample concentration, environment, and instrument response, making it difficult to conduct cross-sectional comparisons of the brightness of different probes; fourth, for fixed-wavelength laser sources, excitation wavelength detuning leads to difficulties in evaluating luminescence brightness. Summary of the Invention

[0005] To address the technical problems mentioned above, this invention provides a method and system for quantifying the luminescence brightness of fluorescent probes with detuned excitation wavelengths. This invention incorporates three dimensions of consideration factors—detuning attenuation weight, light absorption intensity, and radiative transition probability—into the same quantitative index, constructing a comprehensive evaluation parameter that can fully characterize the luminescence brightness of the probe. This provides scientific, efficient, and repeatable technical support for the accurate screening and engineering design of high-brightness probes.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] The first aspect of the present invention provides a method for quantifying the luminescence brightness of a fluorescent probe with detuned excitation wavelength.

[0008] A method for quantifying the luminescence brightness of a fluorescent probe with detuned excitation wavelength includes:

[0009] The initial structure of the probe molecule is constructed, and the geometry of the probe molecule in its ground state is optimized to obtain a stable configuration of the probe molecule in its ground state.

[0010] Based on the stable configuration of the probe molecule's ground state, the excited states are calculated to obtain the probe molecule's lowest ground state. n The excitation energy of each excited state and the excited state in X , Y , Z Absorption transition dipole moments in three directions;

[0011] Based on the stable configuration of the probe molecule's ground state, the geometry of the probe molecule's lowest excited state is optimized to obtain the emission energy and the lowest excited state's energy at that state. X , Y , Z The launch transition dipole moments in three directions;

[0012] Based on the lowest probe molecule n The excitation energy of an excited state, the excited state in X , Y , Z Absorption transition dipole moments in three directions, emission energies of the lowest excited state of the probe molecule, and the lowest excited state in... X , Y , Z Calculate the luminous intensity index based on the emission transition dipole moments in three directions;

[0013] The probe molecule with the highest luminescence index was selected as the optimal probe.

[0014] Furthermore, the initial structure of the probe molecule is constructed, and the geometry of the probe molecule's ground state is optimized to obtain a stable configuration of the probe molecule's ground state; the methods include:

[0015] The initial structure of the probe molecule was constructed using chemical structure drawing software;

[0016] Quantum chemical calculations were used to optimize the geometry of the ground state of the constructed probe molecule, thereby obtaining a stable configuration of the probe molecule's ground state.

[0017] Furthermore, based on the stable configuration of the probe molecule's ground state, the excited states are calculated to obtain the probe molecule's lowest ground state. n The excitation energy of each excited state and the excited state in X , Y , Z Absorption transition dipole moments in three directions; methods include:

[0018] Based on the stable configuration of the probe molecule's ground state, the excited state is calculated using the time-dependent density functional method;

[0019] Extract the lowest value from the excited state calculation results. n Excitation energy of an excited state and minimum n An excited state in X , Y , Z Absorption transition dipole moments in three directions .

[0020] Furthermore, based on the stable configuration of the probe molecule's ground state, the geometry of the probe molecule's lowest excited state is optimized to obtain the emission energy of the lowest excited state and the state of the lowest excited state. X , Y , Z Emission transition dipole moments in three directions; methods include:

[0021] Based on the stable configuration of the probe molecule's ground state, the time-dependent density functional method is used to optimize the geometry of the probe molecule's lowest excited state.

[0022] Extracting the emission energy of the lowest excited state from the results of the lowest excited state structure optimization. and the lowest excited state in X , Y , Z Launch transition dipole moments in three directions .

[0023] Furthermore, based on the lowest level of probe molecules n The excitation energy of an excited state, the excited state in X , Y , Z Absorption transition dipole moments in three directions, emission energies of the lowest excited state of the probe molecule, and the lowest excited state in... X , Y , Z The emission transition dipole moments in three directions are used to calculate the luminance index; the methods include:

[0024] With the lowest probe molecule n Excitation energy of an excited state Construct a diagonal matrix with diagonal elements. ;

[0025] With the first n An excited state in X , Y , Z Absorption transition dipole moments in three directions The square of is the first n Column matrix elements, construct the absorption transition dipole moment matrix ;

[0026] Calculate the dipole moment matrix of the absorption transition D With diagonal matrix E The product of these two elements yields a temporary matrix. ;

[0027] Calculate the temporary matrix DE No. n Columns of columns and with A n For the first n Components establish absorption intensity vector ;

[0028] At the lowest excited state X , Y , Z Launch transition dipole moments in three directions As a component, construct the launch transition dipole moment vector. And calculate the magnitude square of the launch transition dipole moment vector. ;

[0029] Calculate the emission energy of the lowest excited state. cubed With the square of the modulus of the launch transition dipole moment vector To calculate the product, F factor ;

[0030] Based on the proposed excitation wavelength Half-height and full-width and minimum n Excitation energy of an excited state Calculate the correction factor parameters and with correction factor parameters As components, the correction factor vector is obtained. ;

[0031] Calculate the absorption intensity vector A and correction factor vector G inner product and the resulting inner product with F Multiplying the factors yields the luminance index. .

[0032] Furthermore, the luminescence brightness index is positively correlated with the luminescence brightness of the probe molecule; the larger the luminescence brightness index, the higher the luminescence brightness of the probe molecule.

[0033] A second aspect of the present invention provides a fluorescence probe luminance quantification system for coupled excitation wavelength detuning.

[0034] A fluorescence probe luminance quantification system for coupled excitation wavelength detuning includes:

[0035] The configuration optimization module is configured to: construct the initial structure of the probe molecule and optimize the geometry of the probe molecule's ground state to obtain a stable configuration of the probe molecule's ground state;

[0036] The first parameter acquisition module is configured to: calculate the excited state based on the stable configuration of the probe molecule's ground state, and obtain the probe molecule's minimum... n The excitation energy of each excited state and the excited state in X , Y , Z Absorption transition dipole moments in three directions;

[0037] The second parameter acquisition module is configured to: optimize the geometry of the lowest excited state of the probe molecule based on the stable configuration of the probe molecule's ground state, and obtain the emission energy and the lowest excited state's position in the ground state. X , Y , Z The launch transition dipole moments in three directions;

[0038] The brightness index calculation module is configured to: calculate based on the lowest brightness index of the probe molecule. n The excitation energy of an excited state, the excited state in X , Y , Z Absorption transition dipole moments in three directions, emission energies of the lowest excited state of the probe molecule, and the lowest excited state in... X , Y , Z Calculate the luminous intensity index based on the emission transition dipole moments in three directions;

[0039] The prediction and screening module is configured to select the probe molecule with the highest luminescence index as the optimal probe.

[0040] A third aspect of the present invention provides a computer device comprising:

[0041] A processor, adapted to execute computer programs;

[0042] A computer-readable storage medium storing a computer program that, when executed by a processor, implements the steps in the method for quantifying the luminescence brightness of a fluorescent probe with detuned coupled excitation wavelength as described in the first aspect above.

[0043] A fourth aspect of the present invention provides a computer-readable storage medium storing a computer program adapted to be loaded by a processor and to execute steps in the method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength as described in the first aspect above.

[0044] The fifth aspect of the present invention provides a computer program product or computer program.

[0045] This invention provides a computer program product or computer program comprising computer instructions stored in a computer-readable storage medium. A processor of a computer device reads the computer instructions from the computer-readable storage medium and executes the computer instructions, causing the computer device to perform the steps in the method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength as described in the first aspect above.

[0046] Compared with the prior art, the beneficial effects of the present invention are:

[0047] This invention discloses a method and system for quantifying the luminescence brightness of a fluorescent probe with detuned excitation wavelength. By fully considering key parameters such as light absorption intensity and radiative transition rate, a correction factor vector is introduced to describe the comprehensive impact of detuning on probe luminescence brightness, thereby establishing a quantitative mapping relationship between probe energy parameters, transition parameters, and luminescence brightness. Multiple dimensions of factors, including light absorption, light radiation, and excitation source, are integrated into a calculable response function, enabling accurate and rapid prediction of probe luminescence brightness.

[0048] This invention avoids cumbersome and time-consuming processes such as sample preparation, experimental operation, and data acquisition. It not only optimizes the characterization methods of probe luminescence brightness and improves the efficiency and accuracy of brightness calculation, but also has broad applicability and scalability. It can provide an effective tool for the development of biofluorescence imaging technology and help promote the performance comparison and high-throughput screening of fluorescent probes in the field of biomedical imaging. Attached Figure Description

[0049] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0050] Figure 1 This is a flowchart illustrating the method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength, as shown in an embodiment of the present invention.

[0051] Figure 2 This is a flowchart of another embodiment of the fluorescence brightness quantification method for coupled excitation wavelength detuning shown in the embodiments of the present invention;

[0052] Figure 3 This is a schematic diagram of the structure of Chem1 shown in an embodiment of the present invention;

[0053] Figure 4 This is the stable configuration of the Chem1 ground state shown in the embodiments of the present invention;

[0054] Figure 5 This is a schematic diagram of the structure of Chem2 shown in an embodiment of the present invention;

[0055] Figure 6 This is the stable configuration of the Chem2 ground state shown in the embodiments of the present invention;

[0056] Figure 7 This is a schematic diagram of the structure of Chem3 shown in an embodiment of the present invention;

[0057] Figure 8 This is the stable configuration of the Chem3 ground state shown in the embodiments of the present invention;

[0058] Figure 9 This is a schematic diagram of the structure of Chem4 shown in an embodiment of the present invention;

[0059] Figure 10 This is the stable configuration of the Chem4 ground state shown in the embodiments of the present invention;

[0060] Figure 11 This is a structural diagram of a fluorescent probe luminescence brightness quantization system with detuned coupling excitation wavelength, as shown in an embodiment of the present invention.

[0061] Figure 12 This is a structural diagram of a computer device shown in an embodiment of the present invention. Detailed Implementation

[0062] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0063] It should be noted that the following detailed description is illustrative and intended to provide further explanation of the invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0064] It should be noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of exemplary embodiments according to the invention. As used herein, the singular form is intended to include the plural form as well, unless the context clearly indicates otherwise. Furthermore, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0065] To facilitate understanding of the technical solutions of this invention, some technical terms involved in this invention will be introduced below.

[0066] Terminology Explanation: Fluorescent probes are molecules that emit fluorescence when excited by light of a specific wavelength. Their fluorescence signal changes in real time with changes in target ions, enzyme activity, or the microenvironment. They can be used to convert intangible information at the physiological, pathological, or molecular level into visible fluorescent signals, playing an important role in biomedical imaging. Excitation wavelength mistuning refers to the difference between the excitation wavelength and the maximum absorption wavelength; the amount of offset between the two is called the mistuning.

[0067] Figure 1 This is a flowchart illustrating a method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength, as shown in an embodiment of the present invention; see reference. Figure 1 The method includes:

[0068] Step 101: Construct the initial structure of the probe molecule and optimize the geometry of the probe molecule's ground state to obtain a stable configuration of the probe molecule's ground state; specifically including:

[0069] Step 1011: Construct the initial structure of the probe molecule using chemical structure drawing software;

[0070] Step 1012: Optimize the geometric structure of the ground state of the constructed probe molecule using quantum chemical calculation methods to obtain a stable configuration of the probe molecule ground state.

[0071] Step 102: Based on the stable configuration of the probe molecule's ground state, obtain the probe molecule's front state through excited state calculations. n The excitation energy of each excited state and its... X , Y , Z Absorption transition dipole moments in three directions; specifically including:

[0072] Step 1021: Based on the stable configuration of the probe molecule's ground state obtained through optimization, the excited state is calculated using the time-dependent density functional method;

[0073] Step 1022: Extract the lowest value from the excited state calculation results n ( n Excitation energies of ≥ 3 excited states ;

[0074] Step 1023: Extract the lowest value from the excited state calculation results n ( n ≥ 3) excited states in X , Y , Z Absorption transition dipole moments in three directions .

[0075] Step 103: Based on the stable configuration of the probe molecule's ground state, optimize the geometry of the probe molecule's lowest excited state to obtain the emission energy of the probe molecule's lowest excited state and its... X , Y , Z The launch transition dipole moments in three directions; specifically including:

[0076] Step 1031: Based on the stable configuration of the probe molecule's ground state, the time-dependent density functional method is used to optimize the geometry of the probe molecule's lowest excited state;

[0077] Step 1032: Extract the emission energy of the lowest excited state from the results of the lowest excited state structure optimization. ;

[0078] Step 1033: Extract the lowest excited state from the results of the lowest excited state structure optimization. X , Y , Z Launch transition dipole moments in three directions .

[0079] Step 104: Calculate the luminance index by combining the energy parameters and the transition dipole moment parameters; specifically including:

[0080] Step 1041: To stimulate energy Construct a diagonal matrix for the diagonal elements. ;

[0081] Step 1042: Using the first n Excited state in X , Y , Z Absorption transition dipole moments in three directions The square of is the first n The matrix elements of the column are constructed in 3 rows. n The absorption transition dipole moment matrix of the column ;

[0082] Step 1043: Apply the absorption transition dipole matrix D With diagonal matrix E Multiplying yields a temporary matrix. ;

[0083] Step 1044: Calculate the temporary matrix DE No. n Columns of columns and with A n For the first n Components establish absorption intensity vector ;

[0084] Step 1045: Using the launch transition dipole moment As a component, construct the launch transition dipole moment vector And calculate its modulus. ;

[0085] Step 1046: Emitting energy cubed and the launch transition dipole moment vector The model As the multiplier, calculate F factor ;

[0086] Step 1047: Based on the proposed excitation wavelength Half-height and full-width and minimum n ( n Excitation energies of ≥ 3 excited states Calculate the correction factor parameters and with Obtain the correction factor vector for the components. ;

[0087] Step 1048: Calculate the absorption intensity vector A and correction factor vector G inner product and with F Multiplying the factors yields the luminance index. .

[0088] Step 105: Evaluate the luminescence intensity of the probe molecule based on the above index. The higher the index value, the higher the intensity. The probe molecule with the highest luminescence intensity index is selected as the optimal probe. Specifically, this includes:

[0089] Based on luminance index m The numerical value predicts the luminescence intensity of the probe molecule; a larger index value indicates higher luminescence intensity. The optimal luminescence intensity index is... m The largest probe molecule is the brightest probe.

[0090] This invention incorporates three dimensions of factors—detuning attenuation weight, light absorption intensity, and radiative transition probability—into the same quantitative index to construct a comprehensive evaluation parameter that can fully characterize the luminescence brightness of a probe. This provides scientific, efficient, and repeatable technical support for the accurate screening and engineering design of high-brightness probes.

[0091] Figure 2 This is a flowchart of another embodiment of the method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength, as shown in the embodiments of the present invention; in this embodiment, the probe molecule is named Chem1, and the structure of Chem1 is as follows: Figure 3As shown. The method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength includes the following steps:

[0092] Step 201: Construct the initial structure of the probe molecule and optimize the geometry of the probe molecule's ground state to obtain a stable configuration of the probe molecule's ground state. Specifically, this includes:

[0093] Step 2011: Construct the initial structure of the probe molecule using the chemical structure drawing software GaussView 6.0;

[0094] Step 2012: In Gaussian 16 software, the geometry of the ground state of the constructed probe molecule was optimized using the PBE0 functional and the 6-31G(d,p) basis set.

[0095] Step 2013: Open the optimized output file with GaussView 6.0. The displayed structure is the stable ground state configuration of the probe molecule (see...). Figure 4 ).

[0096] Step 202: Based on the stable configuration of the probe molecule's ground state, obtain the probe molecule's front state through excited state calculations. n The excitation energy of each excited state and its... X , Y , Z The absorption transition dipole moments in three directions include:

[0097] Step 2021: Based on the stable configuration of the probe molecule ground state obtained by optimization, excited state calculations containing 8 excited states are performed using the PBE0 functional and the 6-31G(d,p) basis set. The input file keyword is set to "td(nstates=8)pbe1pbe / 6-31g(d,p)".

[0098] Step 2022: From the excited state calculation results, find “Excited State1:”, “Excited State2:”, “Excited State3:”, “Excited State4:”, “Excited State5:”, “Excited State6:”, “Excited State7:”, and “Excited State8:” respectively. The values ​​following these values ​​are the excitation energies of each excited state. E 1 = 3.2902 eV E 2 = 3.6040 eV E 3 = 3.8081 eV E 4 = 4.0033 eV E 5 = 4.2612 eV E6 = 4.4915 eV E 7 = 4.5389 eV E 8 = 4.8819 eV (see Table 1);

[0099] Table 1. Excitation energies of various excited states of the probe molecule Chem1 E n (Unit: eV) In X Absorption transition dipole moment in the direction D nX (Unit: au) Y Absorption transition dipole moment in the direction D nY (Unit: au) Z Absorption transition dipole moment in the direction D nZ (Unit: au)

[0100]

[0101] Step 2023: Find "Ground to excited state transitionelectric dipole moments (Au):" in the excited state calculation results. The value following it is the value of each excited state in the transitionelectric dipole moment (Au): X , Y , Z The absorption transition dipole moments in the three directions are as follows (see Table 1):

[0102]

[0103] Step 203: Based on the stable configuration of the probe molecule's ground state, optimize the geometry of the probe molecule's lowest excited state to obtain the emission energy of the probe molecule's lowest excited state and its... X , Y , Z The launch transition dipole moments in three directions include:

[0104] Step 2031: Based on the stable configuration of the probe molecule's ground state, the lowest excited state geometry of the probe molecule is optimized using the PBE0 functional and the 6-31G(d, p) basis set;

[0105] Step 2032: Find the last "Excited State1:" in the results of the lowest excited state structure optimization. The value following it is the emission energy of the lowest excited state. E' =3.0660 eV (see Table 2);

[0106] Step 2033: Locate the last "Ground to excited state transition electric dipole moments (Au):" in the results of the lowest excited state structure optimization. The value following this is the lowest excited state in the... X , Y , Z The launch transition dipole moments in the three directions are respectively , , (See Table 2).

[0107] Table 2. Emission energies of the lowest excited state of the probe molecule Chem1 E' (Unit: eV) In X Emission transition dipole moment in the direction D' X (Unit: au) Y Emission transition dipole moment in the direction D' Y (Unit: au) Z Emission transition dipole moment in the direction D' Z (Unit: au)

[0108]

[0109] Step 204: Calculate the luminance index by combining the energy parameters and transition dipole moment parameters, specifically including:

[0110] Step 2041: To stimulate energy Construct a diagonal matrix for the diagonal elements. ;

[0111] Step 2042: Using the first n Excited state in X , Y , Z Absorption transition dipole moments in three directions The square of is the first n The matrix elements of the column are constructed in 3 rows. n The absorption transition dipole moment matrix of the column ;

[0112] Step 2043: Calculate the absorption transition dipole moment matrix. D and diagonal matrix E Multiplying yields a temporary matrix. ;

[0113] Step 2044: Calculate the temporary matrix DE No. n Columns of columns A1 = 29.8778, A 2 = 0.0019, A 3 = 0.4539, A 4 = 0.6262, A 5 = 2.1550, A 6 = 5.5690, A 7 = 0.9605, A 8 = 0.6875, and with A n For the first n Components establish absorption intensity vector A = (29.8778, 0.0019, 0.4539, 0.6262, 2.1550, 5.5690, 0.9605, 0.6875);

[0114] Step 2045: Launch the transition dipole moment As a component, construct the launch transition dipole moment vector D' =(3.2269, -0.2694, 0.0008), and calculate its modulus. 3.2269 2 +(-0.2694) 2 +0.0008 2 =10.4855;

[0115] Step 2046: Activate energy cubed and the launch transition dipole moment vector The model As the multiplier, calculate F factor F = (3.0660) 3 ×10.4855 = 302.2066;

[0116] Step 2047: When using the excitation wavelength 405 nm, half-width at half-maximum When the value is 50 nm, the correction factor parameter , , , , , , , The correction factor vector is obtained. G = (0.7597, 0.4024, 0.2841, 0.2160, 0.1613, 0.1307, 0.1258, 0.0988);

[0117] Step 2048: Calculate the absorption intensity vector A and correction factor vector G inner product A·G = 29.8778×0.7597+0.0019×0.4024+0.4539×0.2841+0.6262×0.2160+2.1550×0.1613+5.5690×0.1307+0.9605×0.1258+0.6875×0.0988=24.2267, and combine it with... F Multiplying the factors yields the luminance index. m =( A·G ) × F= 24.2267 × 302.2066 = 7321.4686.

[0118] Step 205: Evaluate the luminescence brightness of the probe molecule based on the above index. The larger the index value, the higher the brightness. The probe molecule with the largest luminescence brightness index is selected as the optimal probe. Specifically, the luminescence index of probe molecule Chem1 is 7321.4686.

[0119] Figure 2 This is a flowchart of another embodiment of the method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength, as shown in the embodiments of the present invention; in this embodiment, the probe molecule is named Chem2, and the structure of Chem2 is as follows: Figure 5 As shown. The method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength includes the following steps:

[0120] Step 301: Construct the initial structure of the probe molecule and optimize the geometry of the probe molecule's ground state to obtain a stable configuration of the probe molecule's ground state. Specifically, this includes:

[0121] Step 3011: Construct the initial structure of the probe molecule using the chemical structure drawing software GaussView 6.0;

[0122] Step 3012: In Gaussian 16 software, the ground-state geometry of the constructed probe molecule is optimized using the PBE0 functional and the 6-31G(d,p) basis set;

[0123] Step 3013: Open the optimized output file with GaussView 6.0. The displayed structure is the stable ground state configuration of the probe molecule (see...). Figure 6 ).

[0124] Step 302: Based on the stable configuration of the probe molecule's ground state, obtain the probe molecule's front state through excited state calculations. n The excitation energy of each excited state and its... X ,Y , Z The absorption transition dipole moments in three directions include:

[0125] Step 3021: Based on the stable configuration of the probe molecule ground state obtained by optimization, excited state calculations containing 8 excited states are performed using the PBE0 functional and the 6-31G(d,p) basis set. The input file keyword is set to "td(nstates=8)pbe1pbe / 6-31g(d,p)";

[0126] Step 3022: From the excited state calculation results, find “Excited State1:”, “Excited State2:”, “Excited State3:”, “Excited State4:”, “Excited State5:”, “Excited State6:”, “Excited State7:”, and “Excited State8:” respectively. The values ​​following these values ​​are the excitation energies of each excited state. E 1 = 3.7013 eV E 2 = 3.7901 eV E 3 = 3.8655 eV E 4 = 3.9799 eV E 5 = 4.2487 eV E 6 = 4.3931 eV E 7 = 4.6624 eV E 8 = 4.7122 eV (see Table 3);

[0127] Step 3023: Find "Ground to excited state transitionelectric dipole moments (Au):" in the excited state calculation results. The value following it is the value of each excited state in the transition electric dipole moments (Au). X , Y , Z The absorption transition dipole moments in the three directions are as follows (see Table 3):

[0128]

[0129] Table 3 Excitation energies of various excited states of the probe molecule Chem2 E n (Unit: eV) In X Absorption transition dipole moment in the direction D nX (Unit: au) Y Absorption transition dipole moment in the direction DnY (Unit: au) Z Absorption transition dipole moment in the direction D nZ (Unit: au)

[0130]

[0131] Step 303: Based on the stable configuration of the probe molecule's ground state, optimize the geometry of the probe molecule's lowest excited state to obtain the emission energy of the probe molecule's lowest excited state and its... X , Y , Z The launch transition dipole moments in three directions include:

[0132] Step 3031: Based on the stable configuration of the probe molecule's ground state, the lowest excited state geometry of the probe molecule is optimized using the PBE0 functional and the 6-31G(d, p) basis set;

[0133] Step 3032: Find the last "Excited State1:" in the results of the lowest excited state structure optimization. The value following it is the emission energy of the lowest excited state. =3.2987 eV (see Table 4);

[0134] Step 3033: Locate the last "Ground to excited state transition electric dipole moments (Au):" in the results of the lowest excited state structure optimization. The value following this is the lowest excited state in the... X , Y , Z The launch transition dipole moments in the three directions are respectively , , (See Table 4).

[0135] Table 4. Emission energies of the lowest excited state of the probe molecule Chem2 E' (Unit: eV) In X Emission transition dipole moment in the direction D' X (Unit: au) Y Emission transition dipole moment in the direction D' Y (Unit: au) Z Emission transition dipole moment in the direction D' Z (Unit: au)

[0136]

[0137] Step 304: Calculate the luminance index by combining the energy parameters and transition dipole moment parameters, specifically including:

[0138] Step 3041: To stimulate energy Construct a diagonal matrix for the diagonal elements. ;

[0139] Step 3042: Using the first n Excited state in X , Y , Z Absorption transition dipole moments in three directions The square of is the first n The matrix elements of the column are constructed in 3 rows. n The absorption transition dipole moment matrix of the column ;

[0140] Step 3043: Calculate the absorption transition dipole moment matrix. D and diagonal matrix E Multiplying yields a temporary matrix. ;

[0141] Step 3044: Calculate the temporary matrix DE No. n Columns of columns A 1 = 5.0399, A 2 = 0.0035, A 3 = 0.0102, A 4 = 3.0669, A 5 = 0.2990, A 6 = 0.4428, A 7 = 0.0224, A 8 = 0.0012, and with A n For the first n Components establish absorption intensity vector A = (5.0399, 0.0035, 0.0102, 3.0669, 0.2990, 0.4428, 0.0224, 0.0012);

[0142] Step 3045: Using the launch transition dipole moment As a component, construct the launch transition dipole moment vector D' =(-1.1215, 0.5814, 0.2131), and calculate its modulus. (-1.1215) 2 +0.5814 2 +0.2131 2 =1.6412;

[0143] Step 3046: Emit energy cubed and the launch transition dipole moment vector The model As the multiplier, calculate F factor F = (3.2987) 3 ×1.6412 = 58.9101;

[0144] Step 3047: When using the excitation wavelength 405 nm, half-width at half-maximum When the value is 50 nm, the correction factor parameter , , , , , , , The correction factor vector is obtained. G = (0.3379, 0.2921, 0.2606, 0.2226, 0.1634, 0.1423, 0.1145, 0.1105);

[0145] Step 3048: Calculate the absorption intensity vector A and correction factor vector G inner product A·G = 5.0399×0.3379+0.0035×0.2921+0.0102×0.2606+3.0669×0.2226+0.2990×0.1634+0.4428×0.1423+0.0224×0.1145+0.0012×0.1105=2.5042, and combine it with... F Multiplying the factors yields the luminance index. m = ( A· G ) × F= 2.5042 × 58.9101 = 147.5227.

[0146] Step 305: Evaluate the luminescence brightness of the probe molecule based on the above index. The larger the index value, the higher the brightness. The probe molecule with the largest luminescence brightness index is preferred as the optimal probe. Specifically, the luminescence index of probe molecule Chem2 is 147.5227, which is lower than the luminescence index of probe molecule Chem1 (7321.4686). Therefore, compared with the two probe molecules, Chem1 is the preferred probe.

[0147] Figure 2This is a flowchart of another embodiment of the method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength, as shown in this invention. In this embodiment, the two probe molecules are named Chem3 and Chem4, and the structure of Chem3 is as follows: Figure 7 As shown, the structure of Chem4 is as follows: Figure 9 As shown. The method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength includes the following steps:

[0148] Step 401: Construct the initial structures of probe molecules Chem3 and Chem4 respectively, and optimize the ground-state geometry of probe molecules Chem3 and Chem4 to obtain the stable ground-state configuration of the probe molecules. Specifically, this includes:

[0149] Step 4011: Construct the initial structures of probe molecules Chem3 and Chem4 using the chemical structure drawing software GaussView 6.0;

[0150] Step 4012: In Gaussian 16 software, the ground state geometry of the constructed probe molecules Chem3 and Chem4 is optimized using the PBE0 functional and the 6-31G(d,p) basis set, respectively.

[0151] Step 4013: Open the output files of the optimized probe molecules Chem3 and Chem4 using GaussView 6.0. The displayed structures are the stable ground-state configurations of the two probe molecules. The stable ground-state configuration of Chem3 is as follows: Figure 8 As shown, the stable configuration of Chem4 in its ground state is as follows: Figure 10 As shown.

[0152] Step 402: Based on the stable ground state configurations of probe molecules Chem3 and Chem4, obtain the respective excitation states of probe molecules Chem3 and Chem4 through excited state calculations. n The excitation energy of each excited state and its... X , Y , Z The absorption transition dipole moments in three directions include:

[0153] Step 4021: Based on the stable ground state configurations of the probe molecules Chem3 and Chem4 obtained through optimization, excited state calculations involving 5 excited states are performed using the PBE0 functional and the 6-31G(d, p) basis set. The input file keywords are set to "td(nstates=5) pbe1pbe / 6-31g(d,p)".

[0154] Step 4022: From the excited state calculation results of the probe molecule Chem3, find “Excited State1:”, “Excited State2:”, “Excited State3:”, “Excited State4:”, and “Excited State5:” respectively. The values ​​following these are the excitation energies of each excited state of Chem3. E 1-Chem3 = 2.7865 eV E 2-Chem3 = 3.0727 eV E 3-Chem3 = 3.4182 eV E 4-Chem3 = 3.7479 eV E 5-Chem3 = 4.1955 eV (see Table 5); Simultaneously, from the excited state calculation results of Chem4, find “Excited State1:”, “Excited State2:”, “Excited State3:”, “Excited State4:”, and “Excited State5:” respectively; the values ​​following these are the excitation energies of each excited state of Chem4, respectively. E 1-Chem4 = 2.7893 eV E 2-Chem4 = 3.0916 eV E 3-Chem4 = 3.4235 eV E 4-Chem4 = 3.8172 eV E 5-Chem4 = 4.1913 eV (see Table 5);

[0155] Table 5. Excitation energies of the excited states of probe molecules Chem3 and Chem4 E n (Unit: eV) In X Absorption transition dipole moment in the direction D nX (Unit: au) Y Absorption transition dipole moment in the direction D nY (Unit: au) Z Absorption transition dipole moment in the direction D nZ (Unit: au)

[0156]

[0157] Step 4023: Find "Ground to excitedstate transition electric dipole moments (Au):" in the excited state calculation results of the probe molecule Chem3. The values ​​following this are the absorption transition dipole moments of each excited state in the X, Y, and Z directions, as shown in Table 5:

[0158]

[0159] Simultaneously, by searching for "Ground to excited state transition electric dipole moments (Au):" in the excited state calculation results of the probe molecule Chem4, the subsequent values ​​represent the excited states at each point in time. X , Y , Z The absorption transition dipole moments in the three directions are as follows (see Table 5):

[0160]

[0161] Step 403: Based on the stable configurations of the ground states of probe molecules Chem3 and Chem4, optimize the geometry of the lowest excited states of the probe molecules to obtain the emission energies of the lowest excited states of Chem3 and Chem4 and their corresponding values. X , Y , Z The launch transition dipole moments in three directions include:

[0162] Step 4031: Based on the stable configuration of the ground state of probe molecules Chem3 and Chem4, the geometry of the lowest excited state of probe molecules Chem3 and Chem4 is optimized using the PBE0 functional and the 6-31G(d, p) basis set, respectively.

[0163] Step 4032: Locate the last "Excited State 1:" in the results of the lowest excited state structure optimization of the probe molecule Chem3. The value following it is the emission energy of the lowest excited state. E' Chem3 =2.0946 eV (see Table 6); simultaneously, the last "Excited State 1:" was found in the results of the lowest excited state structure optimization of the probe molecule Chem4, and the value after it is the emission energy of the lowest excited state. E' Chem4 =2.0942 eV (see Table 6);

[0164] Step 4033: Locate the last "Groundto excited state transition electric dipole moments (Au):" in the results of the lowest excited state structure optimization of the probe molecule Chem3. The value following this is the lowest excited state in the... X , Y , Z The launch transition dipole moments in the three directions are respectively (See Table 6); Simultaneously, from the results of the lowest excited state structure optimization of the probe molecule Chem4, find the last "Ground to excited state transition electric dipole moments (Au):", the subsequent values ​​are the lowest excited state in... X , Y , Z The launch transition dipole moments in the three directions are respectively (See Table 6);

[0165] Table 6. Emission energies of the lowest excited states of probe molecules Chem3 and Chem4 E' (Unit: eV) In X Emission transition dipole moment in the direction D' X (Unit: au) Y Emission transition dipole moment in the direction D' Y (Unit: au) Z Emission transition dipole moment in the direction D' Z (Unit: au)

[0166]

[0167] Step 404: Calculate the luminance index by combining the energy parameters and the transition dipole moment parameters, specifically including:

[0168] Step 4041: Using the excitation energy E of the probe molecule Chem3 1-Chem3 E 2-Chem3 E 3-Chem3 E 4-Chem3 E 5-Chem3 Construct a diagonal matrix for the diagonal elements. Simultaneously, using the excitation energy E of the probe molecule Chem4... 1-Chem4 E 2-Chem4 E 3-Chem4 E 4-Chem4 E 5-Chem4 Construct a diagonal matrix for the diagonal elements. ;

[0169] Step 4042: Using probe molecule Chem3 n Excited state in X , Y , Z Absorption transition dipole moments in three directions The square of is the first n The matrix elements of the column are constructed in 3 rows. n The absorption transition dipole moment matrix of the column Simultaneously, using the probe molecule Chem4... n Excited state in X , Y , Z Absorption transition dipole moments in three directions The square of is the first n The matrix elements of the column are constructed in 3 rows. n The absorption transition dipole moment matrix of the column ;

[0170] Step 4043: Calculate the absorption transition dipole moment matrix of the probe molecule Chem3. D Chem3 and diagonal matrix E Chem3 Multiplying these matrices yields a temporary matrix of the probe molecule Chem3. Simultaneously, the absorption transition dipole moment matrix of the probe molecule Chem4 was determined. D Chem4 and diagonal matrix E Chem4 Multiplying these matrices yields a temporary matrix of the probe molecule Chem4. ;

[0171] Step 4044: Calculate the temporary matrix ( DE ) Chem3 No. n Columns of columns A 1-Chem3 =0.0021, A 2-Chem3 =13.3539, A 3-Chem3 =0.6968, A 4-Chem3 =10.6904, A 5-Chem3 =0.0951 and with A n-Chem3 For the first n Components establish absorption intensity vector A Chem3 = (0.0021, 13.3539, 0.6968, 10.6904, 0.0951); Simultaneously, calculate the temporary matrix ( DE ) Chem4 No. n Columns of columns A 1-Chem4 =0.0019, A 2-Chem4 =12.1754, A 3-Chem4 =0.8851, A 4-Chem4 =9.2914, A 5-Chem3 =0.0942 and with A n-Chem4 For the first n Components establish absorption intensity vector A Chem4 = (0.0019, 12.1754, 0.8851, 9.2914, 0.0942);

[0172] Step 4045: Using the emission transition dipole moment of the probe molecule Chem3 As a component, construct the launch transition dipole moment vector D' Chem3 = (0.0228, 0.0239, -0.0033), and calculate its modulus. 0.0228 2 +0.0239 2 +(-0.0033) 2 =0.0011; simultaneously, the emission transition dipole moment of the probe molecule Chem4 is... As a component, construct the launch transition dipole moment vector D' Chem3 = (-0.0071, 0.0313, 0.0027), and calculate its modulus. (-0.0071) 2 +0.0313 2 +0.0027 2 =0.0010;

[0173] Step 4046: Using the emission energy of the probe molecule Chem3 E' Chem3 cubed ( E' Chem3 ) 3 and the launch transition dipole moment vector D' Chem3 The model As a multiplier, calculate the multiplier of the probe molecule Chem3. F factor F Chem3 ,Right now FChem3 = (2.0946) 3 ×0.0011 = 0.0102; Simultaneously, the emission energy of the probe molecule Chem4... E' Chem4 cubed ( E' Chem4 ) 3 and the launch transition dipole moment vector D' Chem4 The model As a multiplier, calculate the multiplier of the probe molecule Chem4. F factor F Chem4 ,Right now F Chem4 =(2.0942) 3 ×0.0010 = 0.0095;

[0174] Step 4047: When using the excitation wavelength 442 nm, half-width at half-maximum (HWHM) The correction factor parameter of the probe molecule Chem3 at 80 nm , , , , Obtain the correction factor vector G Chem3 = (0.9986, 0.8124, 0.5048, 0.3413, 0.2298); Meanwhile, the correction factor parameter of the probe molecule Chem4... , , , , Obtain the correction factor vector G Chem4 = (0.9990, 0.8031, 0.5013, 0.3180, 0.2306);

[0175] Step 4048: Calculate the absorption intensity vector of the probe molecule Chem3. A Chem3 and correction factor vector G Chem3 inner product A Chem3 ·G Chem3 = 0.0021×0.9986+13.3539×0.8124+0.6968×0.5048+10.6904×0.3413+0.0951×0.2298 = 14.8723, and combine it with... F factorF Chem3 Multiply to obtain the luminance index. m = ( A Chem3 · G Chem3 ) × F Chem3 = 14.8723 × 0.0102 = 0.1510; Simultaneously, calculate the absorption intensity vector of the probe molecule Chem4. A Chem4 and correction factor vector G Chem4 inner product A Chem4 ·G Chem4 = 0.0019×0.9990+12.1754×0.8031+0.5013×0.8851+9.2914×0.3180+0.0942×0.2306 = 13.2001, and then... F factor F Chem4 Multiply to obtain the luminance index. m = ( A Chem4 ·G Chem4 ) × F Chem4 = 13.2001 × 0.0095 = 0.1258.

[0176] Step 405: Evaluate the luminescence brightness of the probe molecule based on the above index. The larger the index value, the higher the brightness. The probe molecule with the largest luminescence brightness index is preferred as the optimal probe. Specifically, the luminescence index of probe molecule Chem3 is 0.1510, which is higher than the luminescence index of probe molecule Chem4 (0.1258). Therefore, compared with the two probe molecules, Chem3 is the preferred probe.

[0177] The above combination Figure 1 , Figure 2 The method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength provided in the embodiments of the present invention has been described in detail. Next, the system for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength provided in the embodiments of the present invention will be described in conjunction with the accompanying drawings.

[0178] Figure 11 This is a schematic diagram of the structure of a fluorescent probe luminescence brightness quantization system with detuned coupling excitation wavelength, as shown in an embodiment of the present invention. (Refer to...) Figure 11 The system described in this invention includes:

[0179] The configuration optimization module is configured to: construct the initial structure of the probe molecule and optimize the geometry of the probe molecule's ground state to obtain a stable configuration of the probe molecule's ground state;

[0180] The first parameter acquisition module is configured to: obtain the front-side parameters of the probe molecule based on the stable configuration of the probe molecule's ground state through excited-state calculations. n The excitation energy of each excited state and its... X , Y , Z Absorption transition dipole moments in three directions;

[0181] The second parameter acquisition module is configured to: optimize the geometry of the lowest excited state of the probe molecule based on the stable configuration of the probe molecule's ground state, and obtain the emission energy of the lowest excited state of the probe molecule and its... X , Y , Z The launch transition dipole moments in three directions;

[0182] The luminance index calculation module is configured to calculate the luminance index by combining the energy parameter and the transition dipole moment parameter.

[0183] The prediction and screening module is configured to evaluate the luminescence brightness of probe molecules based on the above index. The larger the index value, the higher the brightness. The probe molecule with the largest luminescence brightness index is selected as the optimal probe.

[0184] The fluorescence luminance quantification system for coupled excitation wavelength detuned according to embodiments of the present invention can correspond to performing the method described in the embodiments of the present invention, and the above and other operations and / or functions of each module of the fluorescence luminance quantification system for coupled excitation wavelength detuned are respectively for realizing Figure 1 For the sake of brevity, the corresponding processes of each method in the code will not be elaborated here.

[0185] See Figure 12The diagram shows the structure of a computer device, which includes a processor, a communication interface, and a computer-readable storage medium. The processor, communication interface, and computer-readable storage medium are connected via a bus or other means. The communication interface is used to receive and send data. The computer-readable storage medium can be stored in the computer device's memory. The computer-readable storage medium stores computer programs, including program instructions, and the processor executes the program instructions stored in the computer-readable storage medium. The processor (or CPU, Central Processing Unit) is the computing and control core of the computer device, adapted to implement one or more instructions, specifically adapted to load and execute one or more instructions to achieve the corresponding steps in the embodiment of the method for quantifying the luminescence brightness of a fluorescent probe with detuned excitation wavelength.

[0186] This embodiment provides a computer-readable storage medium (Memory), which is a memory device in a computer device used to store programs and data. It is understood that the computer-readable storage medium here can include both the built-in storage medium in the computer device and extended storage media supported by the computer device. The computer-readable storage medium provides storage space that stores the processing system of the computer device.

[0187] Furthermore, this storage space also contains one or more instructions suitable for loading and execution by the processor. These instructions can be one or more computer programs (including program code). It should be noted that the computer-readable storage medium here can be high-speed RAM memory or non-volatile memory, such as at least one disk storage device; optionally, it can also be at least one computer-readable storage medium located remotely from the aforementioned processor.

[0188] In one embodiment, the computer-readable storage medium stores one or more instructions; the processor loads and executes one or more instructions stored in the computer-readable storage medium to implement the corresponding steps in the above embodiment of the method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength.

[0189] This embodiment provides a computer program product or computer program that includes computer instructions stored in a computer-readable storage medium. A processor of a computer device reads the computer instructions from the computer-readable storage medium and executes the computer instructions, causing the computer device to perform the corresponding steps in the above-described embodiment of the method for quantifying the luminescence brightness of a fluorescent probe with detuned excitation wavelength.

[0190] Those skilled in the art will understand that embodiments of the present invention can be provided as methods, systems, or computer program products. Therefore, the present invention can take the form of hardware embodiments, software embodiments, or embodiments combining software and hardware aspects. Furthermore, the present invention can take the form of a computer program product embodied on one or more computer-usable storage media (including, but not limited to, disk storage and optical storage) containing computer-usable program code.

[0191] This invention is described with reference to flowchart illustrations and / or block diagrams of methods, apparatus (systems), and computer program products according to embodiments of the invention. It will be understood that each block of the flowchart illustrations and / or block diagrams, and combinations of blocks in the flowchart illustrations and / or block diagrams, can be implemented by computer program instructions. These computer program instructions can be provided to a processor of a general-purpose computer, special-purpose computer, embedded processor, or other programmable data processing apparatus to produce a machine, such that the instructions, which execute via the processor of the computer or other programmable data processing apparatus, generate instructions for implementing the flowchart illustrations and / or block diagrams. Figure 1 One or more processes and / or boxes Figure 1 A device that provides the functions specified in one or more boxes.

[0192] These computer program instructions may also be stored in a computer-readable storage medium that can direct a computer or other programmable data processing device to function in a particular manner, such that the instructions stored in the computer-readable storage medium produce an article of manufacture including instruction means, which are implemented in a process Figure 1 One or more processes and / or boxes Figure 1 The function specified in one or more boxes.

[0193] These computer program instructions may also be loaded onto a computer or other programmable data processing equipment to cause a series of operational steps to be performed on the computer or other programmable equipment to produce a computer-implemented process, thereby providing instructions that execute on the computer or other programmable equipment for implementing the process. Figure 1 One or more processes and / or boxes Figure 1 The steps of the function specified in one or more boxes.

[0194] Those skilled in the art will understand that all or part of the processes in the above embodiments can be implemented by a computer program instructing related hardware. The program can be stored in a computer-readable storage medium, and when executed, it can include the processes of the embodiments of the above methods. The storage medium can be a magnetic disk, optical disk, read-only memory (ROM), or random access memory (RAM), etc.

[0195] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for quantifying the luminescence brightness of a fluorescent probe with detuned excitation wavelength, characterized in that, include: The initial structure of the probe molecule is constructed, and the geometry of the probe molecule in its ground state is optimized to obtain a stable configuration of the probe molecule in its ground state. Based on the stable configuration of the probe molecule's ground state, the excited states are calculated to obtain the probe molecule's lowest ground state. n The excitation energy of each excited state and the excited state in X , Y , Z Absorption transition dipole moments in three directions; Based on the stable configuration of the probe molecule's ground state, the geometry of the probe molecule's lowest excited state is optimized to obtain the emission energy and the lowest excited state's energy at that state. X , Y , Z The launch transition dipole moments in three directions; Based on the lowest probe molecule n The excitation energy of an excited state, the excited state in X , Y , Z Absorption transition dipole moments in three directions, emission energies of the lowest excited state of the probe molecule, and the lowest excited state in... X , Y , Z The emission transition dipole moments in three directions are used to calculate the luminance index, specifically including: With the lowest probe molecule n Excitation energy of an excited state Construct a diagonal matrix with diagonal elements. ; With the first n An excited state in X , Y , Z Absorption transition dipole moments in three directions The square of is the first n Column matrix elements, construct the absorption transition dipole moment matrix ; Calculate the dipole moment matrix of the absorption transition D With diagonal matrix E The product of these two elements yields a temporary matrix. ; Calculate the temporary matrix DE No. n Columns of columns and with A n For the first n Components establish absorption intensity vector ; At the lowest excited state X , Y , Z Launch transition dipole moments in three directions As a component, construct the launch transition dipole moment vector. And calculate the magnitude square of the launch transition dipole moment vector. ; Calculate the emission energy of the lowest excited state. cubed With the square of the modulus of the launch transition dipole moment vector To calculate the product, F factor ; Based on the proposed excitation wavelength Half-height and full-width and minimum n Excitation energy of an excited state Calculate the correction factor parameters and with correction factor parameters As components, the correction factor vector is obtained. ; Calculate the absorption intensity vector A and correction factor vector G inner product and the resulting inner product with F Multiplying the factors yields the luminance index. ; The probe molecule with the highest luminescence index was selected as the optimal probe.

2. The method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength according to claim 1, characterized in that, The initial structure of the probe molecule is constructed, and the geometry of the probe molecule's ground state is optimized to obtain a stable ground state configuration; the methods include: The initial structure of the probe molecule was constructed using chemical structure drawing software; Quantum chemical calculations were used to optimize the geometry of the ground state of the constructed probe molecule, thereby obtaining a stable configuration of the probe molecule's ground state.

3. The method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength according to claim 1, characterized in that, Based on the stable configuration of the probe molecule's ground state, the excited states are calculated to obtain the probe molecule's lowest ground state. n The excitation energy of each excited state and the excited state in X , Y , Z Absorption transition dipole moments in three directions; methods include: Based on the stable configuration of the probe molecule's ground state, the excited state is calculated using the time-dependent density functional method; Extract the lowest value from the excited state calculation results. n Excitation energy of an excited state and minimum n An excited state in X , Y , Z Absorption transition dipole moments in three directions .

4. The method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength according to claim 1, characterized in that, Based on the stable configuration of the probe molecule's ground state, the geometry of the probe molecule's lowest excited state is optimized to obtain the emission energy and the lowest excited state's energy at that state. X , Y , Z Emission transition dipole moments in three directions; methods include: Based on the stable configuration of the probe molecule's ground state, the time-dependent density functional method is used to optimize the geometry of the probe molecule's lowest excited state. Extracting the emission energy of the lowest excited state from the results of the lowest excited state structure optimization. and the lowest excited state in X , Y , Z Launch transition dipole moments in three directions .

5. The method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength according to claim 1, characterized in that, The luminescence index is positively correlated with the luminescence brightness of the probe molecule; the larger the luminescence index, the higher the luminescence brightness of the probe molecule.

6. A fluorescence probe luminance quantification system for coupled excitation wavelength detuning, characterized in that, include: The configuration optimization module is configured to: construct the initial structure of the probe molecule and optimize the geometry of the probe molecule's ground state to obtain a stable configuration of the probe molecule's ground state; The first parameter acquisition module is configured to: calculate the excited state based on the stable configuration of the probe molecule's ground state, and obtain the probe molecule's minimum... n The excitation energy of each excited state and the excited state in X , Y , Z Absorption transition dipole moments in three directions; The second parameter acquisition module is configured to: optimize the geometry of the lowest excited state of the probe molecule based on the stable configuration of the probe molecule's ground state, and obtain the emission energy and the lowest excited state's position in the ground state. X , Y , Z The launch transition dipole moments in three directions; The brightness index calculation module is configured to: calculate based on the lowest brightness index of the probe molecule. n The excitation energy of an excited state, the excited state in X , Y , Z Absorption transition dipole moments in three directions, emission energies of the lowest excited state of the probe molecule, and the lowest excited state in... X , Y , Z The emission transition dipole moments in three directions are used to calculate the luminance index, specifically including: With the lowest probe molecule n Excitation energy of an excited state Construct a diagonal matrix with diagonal elements. ; With the first n An excited state in X , Y , Z Absorption transition dipole moments in three directions The square of is the first n Column matrix elements, construct the absorption transition dipole moment matrix ; Calculate the dipole moment matrix of the absorption transition D With diagonal matrix E The product of these two elements yields a temporary matrix. ; Calculate the temporary matrix DE No. n Columns of columns and with A n For the first n Components establish absorption intensity vector ; At the lowest excited state X , Y , Z Launch transition dipole moments in three directions As a component, construct the launch transition dipole moment vector. And calculate the magnitude square of the launch transition dipole moment vector. ; Calculate the emission energy of the lowest excited state. cubed With the square of the modulus of the launch transition dipole moment vector To calculate the product, F factor ; Based on the proposed excitation wavelength Half-height and full-width and minimum n Excitation energy of an excited state Calculate the correction factor parameters and with correction factor parameters As components, the correction factor vector is obtained. ; Calculate the absorption intensity vector A and correction factor vector G inner product and the resulting inner product with F Multiplying the factors yields the luminance index. ; The prediction and screening module is configured to select the probe molecule with the highest luminescence index as the optimal probe.

7. A computer device, characterized in that, A processor, adapted to execute computer programs; A computer-readable storage medium storing a computer program, which, when executed by the processor, implements the steps in the method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength as described in any one of claims 1-5.

8. A computer-readable storage medium, characterized in that, The computer-readable storage medium stores a computer program adapted to be loaded by a processor and to execute the steps in the method for quantifying the luminescence brightness of a fluorescent probe with detuned coupled excitation wavelength as described in any one of claims 1-5.

9. A computer program product, characterized in that, The computer program product includes a computer program that, when executed by a processor, implements the steps in the method for quantifying the luminescence brightness of a fluorescent probe with detuned coupling excitation wavelength as described in any one of claims 1-5.

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