A multi-channel composite imaging method and system under fluorescent bimodal excitation

Abstract

The application discloses a kind of multi-channel composite imaging methods and systems under fluorescent bimodal excitation, it is related to contrast imaging technical field, including steps: respectively through the steady-state / transient X-ray excitation under first wavelength and second wavelength each component content target contrast agent;Obtain absolute fluorescence intensity / fluorescence intensity decay curve under each wavelength steady-state / transient X-ray excitation;According to the ratio between absolute fluorescence intensity under each wavelength as the current combination content target contrast agent in three-dimensional encoding space third direction coordinate value;Fitting fluorescence intensity decay curve under each wavelength obtains first fluorescence lifetime / second fluorescence lifetime, and as three-dimensional encoding space first direction / second direction coordinate value;According to coordinate point is adapted between with Lab color model;According to the adaptation result is carried out fluorescence data acquisition and coded imaging.The application avoids the influence of fluorescence contrast aggregation induction and background fluorescence interference on quantitative calculation analysis.

Description

Technical Field

[0001] This invention relates to the field of contrast imaging technology, specifically to a multi-channel composite contrast imaging method and system under fluorescence dual-modal excitation. Background Technology

[0002] With the steady development of optical technologies such as probe chemistry, nanomaterials, confocal optics, multiphoton excitation, and near-infrared detectors, fluorescence imaging has been widely studied and applied due to its advantages of simplicity, speed, real-time performance, and optical visibility. However, due to the complexity of the sample microenvironment, the aggregation-induced effect of fluorescent contrast agents, and background fluorescence interference, it is impossible to determine the true concentration of contrast agents in the sample, thus making it impossible to quantitatively calculate and analyze sample composition and quantity information using absolute fluorescence intensity. Therefore, it is necessary to introduce other optical parameters to quantitatively solve this problem in sample detection. The relative fluorescence intensity ratio can eliminate the interference of background fluorescence, obtaining a one-dimensional ratio value, while transient fluorescence lifetime is independent of external conditions such as contrast agent concentration and laser intensity, resulting in more stable and accurate data.

[0003] Compared to ultraviolet-visible-near-infrared excitation light, X-rays, due to their high energy and short wavelength, exhibit greater material penetration depth. Simultaneously, negligible spontaneous background fluorescence interference can enhance the accuracy and sensitivity of optical signals, attracting increasing attention in the field of imaging detection. One of the main factors limiting its application development is the lack of nanomaterials possessing X-ray excitation, optical fluorescence emission properties, and good biocompatibility. A cross-scale, multi-mode, and visual material detection contrast agent integrating adhesion spectroscopy and X-ray excited fluorescence imaging is an invention of a nanoscale material detection contrast agent that can be excited by an X-ray tube, emitting at multiple wavelengths (blue, green, red, and near-infrared), with tunable emission. This material enables real-time, realistic, and color imaging of samples, allowing for qualitative analysis of sample morphological information. Summary of the Invention

[0004] To address the impact of fluorescent contrast agent aggregation-induced effects and background fluorescence interference on the quantitative calculation and analysis of sample composition and quantity, this invention proposes a multi-channel composite contrast imaging method under fluorescent dual-modal excitation, comprising the following steps:

[0005] S1: The target contrast agent with different component contents is excited by steady-state X-rays at the first and second wavelengths, respectively;

[0006] S2: Obtain the absolute fluorescence intensity of the target contrast agent under steady-state X-ray excitation at the first and second wavelengths, respectively, at the current component content;

[0007] S3: The ratio between the absolute fluorescence intensities at different wavelengths is used as the third-axis coordinate value of the target contrast agent in the three-dimensional coding space under the current combined content;

[0008] S4: The target contrast agent at each component content is excited by transient X-rays at the first and second wavelengths, respectively;

[0009] S5: Obtain the fluorescence intensity decay curves of the target contrast agent under the first and second wavelength transient X-ray excitation at the current component content;

[0010] S6: Obtain the first fluorescence lifetime by fitting the fluorescence intensity decay curve at the first wavelength using the decay function, and use it as the coordinate value of the target contrast agent in the first direction in the three-dimensional coding space at the current combined content;

[0011] S7: The second fluorescence lifetime is obtained by fitting the fluorescence intensity decay curve at the second wavelength through the decay function, and is used as the coordinate value of the target contrast agent in the second direction in the three-dimensional coding space under the current combined content;

[0012] S8: Adapt the target contrast agent to the Lab color model based on the coordinates of the target contrast agent in the three-dimensional coding space according to the content of each component;

[0013] S9: Based on the adaptation results, acquire and encode fluorescence data for the target contrast agent with known component contents.

[0014] Furthermore, the first wavelength and the second wavelength are in different color gamuts, and the second wavelength is greater than the first wavelength.

[0015] Furthermore, the main component of the target contrast agent is NaAF4:B, wherein A is any one of Y, Zr, Nb, Mo, Tc, Fe, Mn, and Bi, and B is any one of Ce, Nd, Ho, Er, Tm, and Yb.

[0016] Furthermore, the first direction, the second direction, and the third direction are perpendicular to each other.

[0017] Furthermore, in step S7, when fitting using the decay function, the fitting result of the fitting function with a statistical variance closer to 1 is selected.

[0018] This invention also proposes a multi-channel composite contrast imaging system under fluorescence dual-modal excitation, comprising:

[0019] The X-ray emission unit is used to emit steady-state X-rays and transient X-rays at the first and second wavelengths, and to excite the target contrast agent at the content of each component.

[0020] The intensity acquisition unit is used to acquire the absolute fluorescence intensity of the target contrast agent under steady-state X-ray excitation at the first and second wavelengths, respectively, at the current component content, and to obtain the ratio between the absolute fluorescence intensities at different wavelengths;

[0021] The attenuation measurement unit is used to collect the fluorescence intensity attenuation curves of the target contrast agent under the first wavelength and the second wavelength transient X-ray excitation at the current component content, and obtain the first fluorescence lifetime by fitting the fluorescence intensity attenuation curve at the first wavelength through the attenuation function, and obtain the second fluorescence lifetime by fitting the fluorescence intensity attenuation curve at the second wavelength.

[0022] The coordinate transformation unit is used to determine the coordinate value in the third direction based on the ratio between the absolute fluorescence intensities of the target contrast agent at different wavelengths at the current content, with the first fluorescence lifetime as the coordinate value in the first direction and the second fluorescence lifetime as the coordinate value in the second direction.

[0023] The color adaptation unit is used to adapt the target contrast agent to the Lab color model based on the coordinates of the target contrast agent in the three-dimensional coding space at the content of each component.

[0024] The contrast imaging unit is used to acquire and encode fluorescence data of the target contrast agent with known component content based on the adaptation results.

[0025] Furthermore, the first wavelength and the second wavelength are in different color gamuts, and the second wavelength is greater than the first wavelength.

[0026] Furthermore, the main component of the target contrast agent is NaAF4:B, wherein A is any one of Y, Zr, Nb, Mo, Tc, Fe, Mn, and Bi, and B is any one of Ce, Nd, Ho, Er, Tm, and Yb.

[0027] Furthermore, the first direction, the second direction, and the third direction are perpendicular to each other.

[0028] Furthermore, in the attenuation calculation unit, when fitting the attenuation function, the fitting result of the fitting function with a statistical variance closer to 1 is selected.

[0029] Compared with the prior art, the present invention has at least the following beneficial effects:

[0030] (1) The multi-channel composite contrast imaging method and system under fluorescence dual-modal excitation described in this invention constructs the coordinate points of the target contrast agent in the three-dimensional coding space according to the absolute fluorescence intensity and fluorescence intensity decay curve, and performs adaptation between Lab color models, thereby avoiding the influence of fluorescence contrast aggregation-induced effect and background fluorescence interference on sample quantitative calculation and analysis.

[0031] (2) The target contrast agent after excitation with different fluorescence lifetimes and different absolute fluorescence intensities is distinguished by color using the Lab model, so as to obtain clearer and more accurate real-time visualized sample morphology information.

[0032] (3) Due to the use of coordinate encoding and Lab model to convert information in equal quantities, it can cover more diverse morphological information and realize the visualization of digital encoding, providing a theoretical basis and methodological guidance for the application of target contrast agents in the field of imaging analysis. Attached Figure Description

[0033] Figure 1 This is a step-by-step diagram of a multi-channel composite contrast imaging method under fluorescence dual-modal excitation;

[0034] Figure 2 This is a structural diagram of a multi-channel composite contrast imaging system excited by fluorescence dual-modal excitation.

[0035] Figure 3 This is a schematic diagram of the capacity space of the Lab color model. Detailed Implementation

[0036] The following are specific embodiments of the present invention, which are described in conjunction with the accompanying drawings. However, the present invention is not limited to these embodiments.

[0037] Example 1

[0038] To develop a novel method for cross-scale, multi-modal, and visualized material detection of contrast agent materials in sample analysis based on mechanospectroscopy and X-ray excited fluorescence imaging, this invention proposes to establish a dual-modal, multi-channel color coding system that combines steady-state relative fluorescence intensity and transient fluorescence lifetime under X-ray excitation. This system enables high-resolution, high-accuracy, and visualized analytical applications in biological detection and medical diagnostics. Specifically, such as... Figure 1 As shown, this invention proposes a multi-channel composite contrast imaging method under fluorescence dual-modal excitation, comprising the following steps:

[0039] S1: The target contrast agent with different component contents is excited by steady-state X-rays at the first and second wavelengths, respectively;

[0040] S2: Obtain the absolute fluorescence intensity of the target contrast agent under steady-state X-ray excitation at the first and second wavelengths, respectively, at the current component content;

[0041] S3: The ratio between the absolute fluorescence intensities at different wavelengths is used as the third-axis coordinate value of the target contrast agent in the three-dimensional coding space under the current combined content;

[0042] S4: The target contrast agent at each component content is excited by transient X-rays at the first and second wavelengths, respectively;

[0043] S5: Obtain the fluorescence intensity decay curves of the target contrast agent under the first and second wavelength transient X-ray excitation at the current component content;

[0044] S6: Obtain the first fluorescence lifetime by fitting the fluorescence intensity decay curve at the first wavelength using the decay function, and use it as the coordinate value of the target contrast agent in the first direction in the three-dimensional coding space at the current combined content;

[0045] S7: The second fluorescence lifetime is obtained by fitting the fluorescence intensity decay curve at the second wavelength through the decay function, and is used as the coordinate value of the target contrast agent in the second direction in the three-dimensional coding space under the current combined content;

[0046] S8: Adapt the target contrast agent to the Lab color model based on the coordinates of the target contrast agent in the three-dimensional coding space according to the content of each component;

[0047] S9: Based on the adaptation results, acquire and encode fluorescence data for the target contrast agent with known component contents.

[0048] In this embodiment, NaAF4:B is used as the target contrast agent, where A can be any one of Y, Zr, Nb, Mo, Tc, Fe, Mn, and Bi, and B can be any one of Ce, Nd, Ho, Er, Tm, and Yb (it should be noted that although this embodiment only specifies contrast agents of the NaAF4:B series, this method can also be used for contrast agents with fluorescence effects in other combinations of components; the specific chemical combination of contrast agent selected depends on the actual situation). The X-ray generating device uses a continuous / pulsed frequency-tunable X-ray tube. The advantage of using this tube is that it can meet two experimental requirements with a single tube, namely, steady-state emission analysis and acquisition of absolute fluorescence intensity at different wavelengths when tuned to continuous mode, and transient emission analysis and acquisition of attenuation curves when tuned to pulsed mode.

[0049] To ensure the stability and reliability of material data obtained based on fluorescence reactions, and considering the potential for varying fluorescence effects due to differences in the proportion of contrast agent components, this invention proposes an imaging method for fluorescence data based on absolute fluorescence intensity and fluorescence intensity decay curves. This method utilizes different color divisions to display sample morphological information. Furthermore, since fluorescence brightness also reflects certain material morphological information, this invention employs the Lab color model as the final imaging color matching source to obtain more accurate material morphological information.

[0050] It is important to understand the Lab color model (such as...) Figure 3 The color shown is composed of three elements: luminance (L) and color-related values ​​a and b. L represents luminance, a represents the range from magenta to green, and b represents the range from yellow to blue. The value of L ranges from 0 to 100; L = 50 is equivalent to 50% black. The values ​​of a and b both range from +127 to -128, where +127a is red, gradually transitioning to green at -128a; similarly, +127b is yellow, and -128b is blue. All colors are composed of the interaction and variation of these three values.

[0051] As can be seen from the above introduction to the Lab color model, (a, b, L) can be considered as coordinates in a three-dimensional coding space. Therefore, if fluorescence data can be converted into coordinate information, it can be adapted to the Lab color model, thereby classifying each sample's morphological information into colors. Here, this invention considers that X-rays at different wavelengths have different fluorescence intensity decay characteristics. Therefore, it is entirely possible to use two different wavelengths of X-rays to excite the target contrast agent at the current component content, thereby obtaining fluorescence intensity decay curves with different decay characteristics, which can be used as the basis for determining the coordinate points in the first direction (x, corresponding to the first wavelength) and the second direction (y, corresponding to the second wavelength) in the three-dimensional coding space. Of course, fluorescence intensity decay curves alone cannot be directly used as coordinate points; appropriate data conversion is required. This invention uses first-order, second-order, and third-order decay functions ExpDec to fit the fluorescence decay curves under two different wavelengths, and calculates the corresponding fluorescence lifetime values ​​of the target contrast agent at different wavelengths. Preferably, the fluorescence lifetime value is selected as the statistical variance R after fitting. 2 The result obtained by fitting a function that is closer to 1.

[0052] For the coordinate points in the third direction (z) of the three-dimensional coordinate space, this invention obtains them using absolute fluorescence intensity data. Since the coordinate points in the first and second directions are obtained through excitation with two different wavelengths of X-rays, and using only the intensity of one wavelength as the basis for obtaining the coordinate points is clearly unreasonable, we here select the ratio between the absolute fluorescence intensity under the second wavelength X-ray excitation and the absolute fluorescence intensity under the first wavelength X-ray excitation as the coordinate point in the third direction.

[0053] Based on the above, this section uses NaYF4:28%Er as an example to illustrate how to obtain contrast imaging, where 28% represents the current component content, which is the content ratio of Er / (Er+Y). The three-dimensional encoded spatial coordinates (x, y, y) are obtained using the method described above. 28 y 28 , z 28The subscript 28 corresponds to the current component content of 28%. Since the a and b color channel signals in the Lab color model are in the range [127, -128], which differs from the capacity of a conventional three-dimensional encoding space, further processing is required. This processing can be represented as follows: The superscript m here refers to x n or y n The value of , e is the natural constant. And for the absolute fluorescence intensity ratio z n Then proceed The processing, where the superscript m refers to z n The value is set to conform to the value range [0,100] of the L luminance channel in the Lab color model.

[0054] Using the above data processing, capacity matching between the infinite-domain 3D contrast agent coding library and the finite-domain Lab color model space was successfully achieved (e.g., Figure 3 As shown, the subscripts of the three-dimensional coding coordinates in the coding matrix space represent the component content. This completes the establishment of a dual-modal, multi-channel composite fluorescence coding method that transforms the target contrast agent, with the doping content n as the sole variable, into digital coding and then into color matching.

[0055] Example 2

[0056] To better understand the technical content of this invention, this embodiment describes the invention through a system structure, such as... Figure 2 As shown, a multi-channel composite contrast imaging system under fluorescence dual-modal excitation includes:

[0057] The X-ray emission unit is used to emit steady-state X-rays and transient X-rays at the first and second wavelengths, and to excite the target contrast agent at the content of each component.

[0058] The intensity acquisition unit is used to acquire the absolute fluorescence intensity of the target contrast agent under steady-state X-ray excitation at the first and second wavelengths, respectively, at the current component content, and to obtain the ratio between the absolute fluorescence intensities at different wavelengths;

[0059] The attenuation measurement unit is used to collect the fluorescence intensity attenuation curves of the target contrast agent under the first wavelength and the second wavelength transient X-ray excitation at the current component content, and obtain the first fluorescence lifetime by fitting the fluorescence intensity attenuation curve at the first wavelength through the attenuation function, and obtain the second fluorescence lifetime by fitting the fluorescence intensity attenuation curve at the second wavelength.

[0060] The coordinate transformation unit is used to determine the coordinate value in the third direction based on the ratio between the absolute fluorescence intensities of the target contrast agent at different wavelengths at the current content, with the first fluorescence lifetime as the coordinate value in the first direction and the second fluorescence lifetime as the coordinate value in the second direction.

[0061] The color adaptation unit is used to adapt the target contrast agent to the Lab color model based on the coordinates of the target contrast agent in the three-dimensional coding space at the content of each component.

[0062] The contrast imaging unit is used to acquire and encode fluorescence data of the target contrast agent with known component content based on the adaptation results.

[0063] Furthermore, the first wavelength and the second wavelength are in different color gamuts, and the second wavelength is greater than the first wavelength.

[0064] Furthermore, the main component of the target contrast agent is NaAF4:B, where A is any one of Y, Zr, Nb, Mo, Tc, Fe, Mn, and Bi, and B is any one of Ce, Nd, Ho, Er, Tm, and Yb.

[0065] Furthermore, the first direction, the second direction, and the third direction are perpendicular to each other.

[0066] Furthermore, in the attenuation measurement unit, when fitting the attenuation function, the fitting result of the fitting function with a statistical variance closer to 1 is selected.

[0067] In summary, the multi-channel composite contrast imaging method and system under fluorescence dual-modal excitation described in this invention constructs the coordinates of the target contrast agent in a three-dimensional coding space based on the absolute fluorescence intensity and fluorescence intensity decay curve, and performs adaptation between Lab color models, thereby avoiding the influence of fluorescence contrast aggregation-induced effects and background fluorescence interference on the quantitative calculation and analysis of samples.

[0068] By using the Lab model to differentiate and display target contrast agents excited with different fluorescence lifetimes and absolute fluorescence intensities by color, clearer and more accurate real-time visualization of sample morphology information can be obtained.

[0069] By employing an equal conversion of information between coordinate encoding and Lab model, it can encompass more diverse morphological information and achieve visualization of digital encoding, providing a theoretical basis and methodological guidance for the application of target contrast agents in the field of imaging analysis.

[0070] It should be noted that all directional indications (such as up, down, left, right, front, back, etc.) in the embodiments of the present invention are only used to explain the relative positional relationship and movement of each component in a certain specific posture (as shown in the figure). If the specific posture changes, the directional indication will also change accordingly.

[0071] Furthermore, in this invention, descriptions involving terms such as "first," "second," and "a" are for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0072] In this invention, unless otherwise explicitly specified and limited, the terms "connection," "fixed," etc., should be interpreted broadly. For example, "fixed" can mean a fixed connection, a detachable connection, or an integral part; it can mean a mechanical connection or an electrical connection; it can mean a direct connection or an indirect connection through an intermediate medium; it can mean the internal communication of two components or the interaction between two components, unless otherwise explicitly limited. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0073] Furthermore, the technical solutions of the various embodiments of the present invention can be combined with each other, but only if they are feasible for those skilled in the art. If the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such combination of technical solutions does not exist and is not within the scope of protection claimed by the present invention.

Claims

1. A multi-channel composite contrast imaging method under fluorescence dual-modal excitation, characterized in that, Including the following steps: S1: The target contrast agent at each component content is excited by steady-state X-rays at a first wavelength and a second wavelength, respectively. The first wavelength and the second wavelength are in different color gamuts, and the second wavelength is greater than the first wavelength. S2: Obtain the absolute fluorescence intensity of the target contrast agent under steady-state X-ray excitation at the first and second wavelengths, respectively, at the current component content; S3: The ratio between the absolute fluorescence intensities at different wavelengths is used as the third-axis coordinate value of the target contrast agent in the three-dimensional coding space under the current combined content; S4: The target contrast agent at each component content is excited by transient X-rays at the first and second wavelengths, respectively; S5: Obtain the fluorescence intensity decay curves of the target contrast agent under the first and second wavelength transient X-ray excitation at the current component content; S6: Obtain the first fluorescence lifetime by fitting the fluorescence intensity decay curve at the first wavelength using the decay function, and use it as the coordinate value of the target contrast agent in the first direction in the three-dimensional coding space at the current combined content; S7: The second fluorescence lifetime is obtained by fitting the fluorescence intensity decay curve at the second wavelength through the decay function, and is used as the coordinate value of the target contrast agent in the second direction in the three-dimensional coding space under the current combined content. The first direction, the second direction and the third direction are perpendicular to each other. S8: Adapt the target contrast agent to the Lab color model based on the coordinates of the target contrast agent in the three-dimensional coding space according to the content of each component; S9: Based on the adaptation results, acquire and encode fluorescence data for the target contrast agent with known component contents.

2. The multi-channel composite contrast imaging method under fluorescence dual-modal excitation as described in claim 1, characterized in that, The main component of the target contrast agent is NaAF4:B, wherein A is any one of Y, Zr, Nb, Mo, Tc, Fe, Mn, and Bi, and B is any one of Ce, Nd, Ho, Er, Tm, and Yb.

3. The multi-channel composite contrast imaging method under fluorescence dual-modal excitation as described in claim 1, characterized in that, In step S7, when fitting using the decay function, the fitting result of the fitting function with a statistical variance closer to 1 is selected.

4. A multi-channel composite contrast imaging system under fluorescence dual-modal excitation, characterized in that, include: The X-ray emitting unit is used to emit steady-state X-rays and transient X-rays at a first wavelength and a second wavelength, and to excite the target contrast agent at various component contents. The first wavelength and the second wavelength are in different color gamuts, and the second wavelength is greater than the first wavelength. The intensity acquisition unit is used to acquire the absolute fluorescence intensity of the target contrast agent under steady-state X-ray excitation at the first and second wavelengths, respectively, at the current component content, and to obtain the ratio between the absolute fluorescence intensities at different wavelengths; The attenuation measurement unit is used to collect the fluorescence intensity attenuation curves of the target contrast agent under the first wavelength and the second wavelength transient X-ray excitation at the current component content, and obtain the first fluorescence lifetime by fitting the fluorescence intensity attenuation curve at the first wavelength through the attenuation function, and obtain the second fluorescence lifetime by fitting the fluorescence intensity attenuation curve at the second wavelength. The coordinate transformation unit is used to determine the coordinate value in the third direction based on the ratio between the absolute fluorescence intensities of the target contrast agent at different wavelengths at the current content, with the first fluorescence lifetime as the coordinate value in the first direction and the second fluorescence lifetime as the coordinate value in the second direction, wherein the first direction, the second direction and the third direction are perpendicular to each other. The color adaptation unit is used to adapt the target contrast agent to the Lab color model based on the coordinates of the target contrast agent in the three-dimensional coding space at the content of each component. The contrast imaging unit is used to acquire and encode fluorescence data of the target contrast agent with known component content based on the adaptation results.

5. The multi-channel composite contrast imaging system under fluorescence dual-modal excitation as described in claim 4, characterized in that, The main component of the target contrast agent is NaAF4:B, wherein A is any one of Y, Zr, Nb, Mo, Tc, Fe, Mn, and Bi, and B is any one of Ce, Nd, Ho, Er, Tm, and Yb.

6. The multi-channel composite contrast imaging system under fluorescence dual-modal excitation as described in claim 4, characterized in that, In the attenuation measurement unit, when fitting the attenuation function, the fitting result of the fitting function with a statistical variance closer to 1 is selected.

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