Method and apparatus for imaging arterial wall structure

Abstract

The application provides an imaging method and device for an arterial wall structure, a soliton light source signal is obtained by preprocessing a laser signal, the soliton light source signal is emitted to a microscopic imaging system, a first target position of a target object is scanned and imaged by the microscopic imaging system, and a third harmonic image and a three-photon fluorescence image are obtained. According to the technical scheme, the soliton light source signal of a specific wavelength is obtained by preprocessing the laser signal in advance, the soliton light source signal has better adaptability to a marker (stronger excitation characteristic), the imaging quality of the third harmonic image and the three-photon fluorescence image after imaging of the microscopic imaging system is improved, direct visualization and easy resolution of different layers of the arterial wall structure are achieved, and the microscopic imaging system configured with a gallium arsenide photomultiplier tube and a gallium arsenic phosphor photomultiplier tube is used, two-channel acquisition is adopted to realize twice scanning and imaging of the first target position of the target object, and the number of obtained image details is ensured.

Description

Technical Field

[0001] This invention relates to the field of structural imaging technology, specifically to imaging methods and apparatus for arterial wall structures. Background Technology

[0002] Currently, a comprehensive understanding of the structure of brain blood vessels is a prerequisite for conducting research on their physiology and function. This places demands on imaging technology, requiring subcellular resolution, depth of penetration, and optical sectioning capabilities. However, most existing imaging methods rely solely on conventional microscopy. When imaging live organisms, the resulting images suffer from low image quality, low resolution, and limited detail, failing to meet the requirements for relevant research on brain blood vessel physiology and function.

[0003] Therefore, existing technologies need to be improved. Summary of the Invention

[0004] The main objective of this invention is to provide an imaging method and apparatus for arterial wall structure, so as to at least solve the technical problem of low imaging quality in related technologies.

[0005] A first aspect of the present invention provides an imaging method for arterial wall structures, comprising:

[0006] The laser signal is preprocessed to obtain the soliton source signal;

[0007] The soliton light source signal is transmitted to the microscopic imaging system;

[0008] The first target location of the target object is scanned and imaged using the microscopic imaging system to obtain a third harmonic image and a three-photon fluorescence image; wherein the third harmonic image and the three-photon fluorescence image are used to display the arterial wall structure of the target object.

[0009] Based on the first aspect, before the step of preprocessing the laser signal, the method further includes: starting the laser device and connecting the laser device to the filter device, and transmitting a laser signal with a wavelength of 1550nm to the filter device through the laser device.

[0010] Based on the first aspect, the step of preprocessing the laser signal to obtain the soliton source signal specifically includes: filtering the laser signal through a filtering device to generate a soliton source signal with a wavelength of 1665nm.

[0011] Based on the first aspect, the step of transmitting the soliton light source signal to the microscopic imaging system specifically includes: guiding the soliton light source signal to the microscopic imaging system via a signal guiding device; wherein the signal guiding device includes a plurality of lens structures for refracting the soliton light source signal.

[0012] Based on the first aspect, the microscopic imaging system is equipped with a gallium arsenide photomultiplier tube with a 540 / 80nm bandpass filter and a gallium arsenide-phosphorus photomultiplier tube with a 630 / 82nm bandpass filter; the step of scanning and imaging the first target position of the target object through the microscopic imaging system to obtain a third harmonic image and acquire a three-photon fluorescence image specifically includes:

[0013] The aforementioned microscopic imaging system is used to perform imaging scanning at the first target position of the target object;

[0014] The third harmonic image is obtained using the gallium arsenide photomultiplier tube;

[0015] An Alexa fluor633 marker is injected into the second target location of the target object, and a three-photon fluorescence image is acquired through the gallium arsenide phosphide photomultiplier tube.

[0016] Based on the first aspect, after the step of acquiring a three-photon fluorescence image through the gallium arsenide phosphide photomultiplier tube, the method further includes:

[0017] When the reaction time of the Alexa fluor 633 marker is greater than a first preset time, a line graph showing a first marked region is obtained based on the third harmonic image and the three-photon fluorescence image; wherein, the first marked region is used to represent the elastic fiber layer in the arterial wall structure.

[0018] Based on the first aspect, after the step of acquiring the three-photon fluorescence image, the method further includes:

[0019] Inject WGA-594 markers at the second target location of the target object;

[0020] When the reaction time of the WGA-594 marker is greater than a second preset time, a line graph showing the second marker region is obtained based on the third harmonic image and the three-photon fluorescence image; wherein, the second marker region is used to represent vascular endothelial cells in the arterial wall structure.

[0021] Building upon the first aspect, after the step of acquiring the three-photon fluorescence image, the method further includes:

[0022] Inject WGA-594 markers at the second target location of the target object;

[0023] When the reaction time of the WGA-594 marker is greater than a third preset time, a line graph showing a third marker region is obtained based on the third harmonic image and the three-photon fluorescence image; wherein, the third marker region is used to represent the smooth muscle cell layer in the arterial wall structure.

[0024] A second aspect of the present invention provides an imaging device for arterial wall structure, comprising a laser device, a filtering device, and a microscopic imaging system:

[0025] The laser device is used to emit laser signals with a wavelength of 1550nm;

[0026] The filtering device is used to preprocess the laser signal and send the generated soliton source signal to the microscopic imaging system;

[0027] The microscopic imaging system is used to scan and image the first target location of the target object to obtain a third harmonic image and a three-photon fluorescence image; wherein, the third harmonic image and the three-photon fluorescence image are used to display the arterial wall structure of the target object.

[0028] Based on the second aspect, the microscopic imaging system is equipped with a gallium arsenide photomultiplier tube with a 540 / 80nm bandpass filter and a gallium arsenide phosphorus photomultiplier tube with a 630 / 82nm bandpass filter; wherein, the gallium arsenide photomultiplier tube is used to acquire the third harmonic image corresponding to the target object, and the gallium arsenide phosphorus photomultiplier tube is used to acquire the three-photon fluorescence image corresponding to the target object.

[0029] The imaging method and apparatus for arterial wall structure of the present invention have the following beneficial effects:

[0030] 1. A soliton source signal is obtained by preprocessing the laser signal. This soliton source signal is then emitted into a microscopic imaging system, which scans and images the first target position of the object to obtain a third harmonic image and a three-photon fluorescence image. Because the laser signal is preprocessed to obtain a soliton source signal of a specific wavelength, the soliton source signal has better compatibility with the marker (stronger excitation characterization characteristics), thereby improving the imaging quality of the third harmonic image and the three-photon fluorescence image after imaging by the microscopic imaging system. This achieves direct visualization and easy differentiation of different layers of the arterial wall structure.

[0031] 2. A microscopic imaging system equipped with gallium arsenide photomultiplier tubes and gallium arsenide-phosphorus photomultiplier tubes employs dual-channel acquisition technology to simultaneously scan and image the first target location of the object, obtaining corresponding third harmonic images and three-photon fluorescence images. More images reflect more details within the target object, ensuring a high level of detail acquisition. For example, details could include the elastic fiber layer, vascular endothelial cells, and smooth muscle cell layer in the arterial wall structure. Attached Figure Description

[0032] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0033] Figure 1 This is a schematic flowchart of an imaging method for arterial wall structure provided in one embodiment of the present invention;

[0034] Figure 2 This is a schematic diagram of the structural connection between the laser device, the signal guiding device, and the microscopic imaging system in one embodiment of the present invention;

[0035] Figure 3 This is a spectrum of a soliton light source signal in one embodiment of the present invention;

[0036] Figure 4 This is a graph showing the variation trend of fluorescence signal of WGA-594 sample with excitation power when the three-photon fluorescence of Alexa Fluor 633 is excited at 1600nm, 1700nm and 1800nm, respectively, in one embodiment of the present invention.

[0037] Figure 5 This is a schematic diagram of the three-photon interaction cross section of the WGA-594 marker in one embodiment of the present invention;

[0038] Figure 6 This is a three-photon fluorescence image of a blood vessel wall on the brain surface labeled with Alexa fluor633 in one embodiment of the present invention;

[0039] Figure 7 This is a third harmonic image of a blood vessel wall on the brain surface labeled with Alexa fluor633 markers in one embodiment of the present invention;

[0040] Figure 8 For Figure 6 , Figure 7 The line drawing at position 1 in the middle;

[0041] Figure 9 For Figure 6 , Figure 7 The line drawing at position 2 in the middle;

[0042] Figure 10 This is a third harmonic image of a blood vessel when the reaction time of the WGA-594 marker is greater than a second preset time period, according to one embodiment of the present invention.

[0043] Figure 11This is a three-photon fluorescence image of a blood vessel when the reaction time of the WGA-594 marker is greater than a second preset time period, according to one embodiment of the present invention.

[0044] Figure 12 To Figure 10 , Figure 11 The merged image resulting from the merging process;

[0045] Figure 13 for Figure 12 The line graph corresponding to the merged image;

[0046] Figure 14 This is a third harmonic image of a blood vessel when the reaction time of the WGA-594 marker is greater than a third preset time period, according to one embodiment of the present invention.

[0047] Figure 15 To and Figure 14 The corresponding line drawing;

[0048] Figure 16 for Figure 14 A magnified view of the portion highlighted above;

[0049] Figure 17 for Figure 14 A magnified view of the portion highlighted below;

[0050] Figures 18 to 21 Three-photon fluorescence images of smooth muscle cell layers at different depths in the arterial wall after the reaction time for WGA-594 marker is longer than the third preset time period.

[0051] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0052] It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0053] It should be noted that related terms such as "first" and "second" can be used to describe various components, but these terms do not limit the component. These terms are only used to distinguish one component from another. For example, without departing from the scope of the invention, the first component can be referred to as the second component, and the second component can similarly be referred to as the first component. The term "and / or" refers to any one or more combinations of related and descriptive terms.

[0054] In in vivo imaging studies using related technologies, transgenic mice are commonly used to distinguish between vascular smooth muscle cells and endothelial cells, significantly increasing costs. In non-transgenic mice, WGA (Wheat Germ Agglutinin) is commonly used for direct labeling to visualize the endothelial cell layer. However, direct labeling of vascular smooth muscle cells for in vivo imaging has not yet been demonstrated; that is, imaging of the middle cerebral artery wall structure has not been achieved.

[0055] To address the technical challenges of imaging the middle cerebral artery wall structure, which is not achievable in the aforementioned related technologies, please refer to [link to relevant documentation]. Figure 1 and Figure 2 The imaging methods for this arterial wall structure include:

[0056] Step S10: Preprocess the laser signal to obtain the soliton source signal.

[0057] Specifically, the laser device 201 is activated, and the laser device 201 emits a laser signal with a wavelength of 1550nm. This laser signal can be preprocessed, such as filtered, to form a soliton source signal with a wavelength of 1665nm.

[0058] It should be noted that soliton signals, also known as solitary waves, are a special type of ultrashort pulse, or a pulsed traveling wave whose shape, amplitude, and velocity remain constant during propagation. Generally, during frequency shifts, due to the balance between the nonlinear change in refractive index and the group dispersion effect, the optical pulse forms a fundamental soliton, which propagates stably in the anomalous dispersion region. In other words, an optical soliton is a light pulse that can propagate in optical fiber while maintaining its shape, amplitude, and velocity for a long time. The characteristics of optical solitons can be utilized to achieve ultra-long-distance, ultra-high-capacity optical communication.

[0059] Optionally, a signal guiding device is provided between the laser device 201 and the filter device Filter. The signal guiding device includes multiple lens structures for refracting soliton light source signals to guide the laser signal with a wavelength of 1550nm to the filter device Filter.

[0060] Specifically, the signal guiding device includes, in sequence, a half-wave plate (HWP), a polarization beam splitter (PBS), a first lens (L1), a photonic crystal rod fiber (PC Rod), a second lens (L2), a 1635 nm long-pass filter (LPF), a first mirror structure (mirror1), a second mirror structure (XY scan mirrors), a scanning lens, a tube lens, a third mirror structure (mirror2), and a conversion structure (DC). These devices stably transfer a 1550 nm femtosecond laser signal to the filtering device (Filter). After passing through the combination of HWP and PBS, the light source passes through a reflector and is then focused by a focusing lens into the input end of the PC Rod. At the output end, it is collimated by a collimating lens. By switching different filters, different wavelength solitons are output to perform filtering processing.

[0061] Step S20: The soliton light source signal is transmitted to the microscopic imaging system.

[0062] Specifically, because the soliton source signal maintains its shape, amplitude, and speed unchanged for a long time during propagation in the optical fiber, transmitting a soliton source signal with a wavelength of 1665 nm to the microscopic imaging system 202 can greatly ensure the stability of the imaging. The spectrum of the soliton source signal with a wavelength of 1665 nm is as follows: Figure 3 As shown.

[0063] Optionally, the microscopic imaging system 202 is equipped with a gallium arsenide photomultiplier tube with a 540 / 80nm bandpass filter and a gallium arsenide phosphide photomultiplier tube with a 630 / 82nm bandpass filter, which has the advantage of dual-channel acquisition. Different channels can acquire different images, ensuring the amount of signal acquired in the subsequent process.

[0064] Step S30: Scan the first target position of the target object using a microscopic imaging system to obtain a third harmonic image and a three-photon fluorescence image.

[0065] Specifically, since the microscopic imaging system 202 is a dual-channel acquisition system, during imaging scanning, when the microscopic imaging system 202 is aligned with the first target position of the target object, it acquires a third harmonic generation (THG) image through a gallium arsenide photomultiplier tube and a three-photon fluorescence image (3PF) through a gallium arsenide phosphorus photomultiplier tube. The target object can be a non-transgenic mouse (black mouse), and the first target position is the head of the non-transgenic mouse (black mouse).

[0066] It should be noted that three-photon fluorescence and third harmonic generation are collectively referred to as three-photon microscopy. Three-photon fluorescence is a non-parametric process, requiring the simultaneous absorption of three photons to transition the molecule from its ground state to an excited state, followed by a vibrational non-radiative transition, and finally the emission of a fluorescent photon. Third harmonic generation, on the other hand, is a parametric process, not involving real energy levels. It requires phase-matching conditions, and the frequency of the generated signal light must precisely satisfy a multiple relationship with the excitation frequency, i.e., three times. The fluorescence and harmonic signals are detected by photodetectors such as photomultiplier tubes and transmitted to a computer via a data acquisition card to form an image.

[0067] Through the above embodiments, the technical solution of this application preprocesses the laser signal to obtain a soliton light source signal, emits the soliton light source signal to a microscopic imaging system, and scans and images the first target position of the target object through the microscopic imaging system to obtain a third harmonic image and a three-photon fluorescence image. That is, the technical solution of this application, on the one hand, achieves direct labeling of vascular smooth muscle in vivo imaging (imaging of the middle cerebral artery wall structure), which is impossible in related technologies; on the other hand, because the laser signal is preprocessed to obtain a soliton light source signal of a specific wavelength, the soliton light source signal has better compatibility with the label (stronger excitation characterization characteristics), thereby improving the imaging quality of the third harmonic image and the three-photon fluorescence image after imaging by the microscopic imaging system, achieving direct visualization and easy differentiation of different layers of the arterial wall structure.

[0068] It should be further noted that, in some specific measurement processes, the fluorescence signals of the WGA-594 label were measured at different excitation powers of 1600, 1700, and 1800 nm. The measurement results are as follows: Figure 4 As shown, the linear fitting plot of the logarithmically transformed measurement data was used to measure the nonlinear order by obtaining its slope. The fitting slopes of the WGA-594 label at excitation in the 1600, 1700, and 1800 nm wavelengths were 2.95, 2.98, and 3.06, respectively, indicating that the WGA-594 label can generate a three-photon fluorescence signal when excited in the 1700 nm window. Thus, the 3-photon interaction cross section (ησ3) of the WGA-594 label, measured at the 1700 nm wavelength window with SR101 as a reference, has a peak at 1620 nm (e.g., ...). Figure 5 (As shown). Experimental results show that 1665nm solitons have higher pulse energy than 1620nm solitons, and the ησ3 of the two wavelengths are close. Therefore, in this embodiment, 1665nm can be selected as the excitation wavelength for imaging, that is, a soliton source signal with a wavelength of 1665nm.

[0069] In some optional embodiments of this example, after the step of scanning and imaging the first target location of the target object using the microscopic imaging system, the method further includes: injecting the Alexafluor633 marker into the second target location of the target object to obtain a three-photon fluorescence image.

[0070] Specifically, when the reaction time of the Alexa fluor633 marker is greater than a first preset time, a line graph showing the first marker region is obtained based on the third harmonic image and the three-photon fluorescence image.

[0071] Specifically, an Alexa fluor633 marker is injected into the second target location of the target object. When the reaction time of the Alexa fluor633 marker is greater than a first preset time (the first preset time can be 10 seconds), based on the third harmonic image (see [link]). Figure 6 ) and three-photon fluorescence images (see Figure 7 A line drawing showing the first marked area is obtained. Since the third harmonic imaging and three-photon fluorescence imaging clearly show the vessel wall, a line drawing spanning the vessel wall can be drawn to better illustrate their spatial location (see [link to image]). Figure 8 , 9 , Figure 8 and Figure 9 They represent Figure 6 , Figure 7 The line graphs labeled 1 and 2 clearly show the first marker region (the overlapping area between peaks), indicating that the elastic fiber layer on the arterial wall produces a strong THG signal. This could potentially be used to differentiate and display evidence of the anatomical location of ECs and VSMCs, and to represent the elastic fiber layer in the arterial wall structure. The reaction time of the Alexa fluor 633 marker indicates the time interval after the Alexa fluor 633 marker is injected into the target object. Alexa Fluor 633 is a bright and photostable far-infrared fluorescent dye, and the second target location could be the orbit.

[0072] In some optional embodiments of this example, after the step of acquiring the three-photon fluorescence image, the method further includes: injecting a WGA-594 marker into a second target location of the target object; when the reaction time of the WGA-594 marker is greater than a second preset time, obtaining a line graph showing the second marked area based on the third harmonic image and the three-photon fluorescence image.

[0073] Specifically, the second preset time can be 60 minutes; when the reaction time of the WGA-594 marker is greater than 60 minutes, the lumen side of the vessel wall is marked and can be seen in the third harmonic imaging (e.g., Figure 10(as shown); To further determine the location of the blood vessel wall marked by the WGA-594 marker, a three-photon fluorescence image was obtained (as shown). Figure 11 (as shown), and merge the third harmonic image and the three-photon fluorescence image to obtain the merged image (as shown). Figure 12 As shown), and based on the merged image, draw a line graph showing the area marked with the second marker (e.g. Figure 13 As shown in the image, it can be clearly seen that the vascular wall structures shown in the three-photon fluorescence image are on the luminal side compared to the third harmonic image (from the elastic fiber layer), indicating that their signals originate from ECs, which is consistent with previous results that this second marker region is used to represent vascular endothelial cells in the arterial wall structure.

[0074] It should be noted that, for the WGA-594 label, to ensure clear labeling of the three-photon fluorescence image, a three-photon interaction cross-section (ησ3) measurement system was constructed. This system outputs a 1-MHz femtosecond pulse with a wavelength tuned from 1600nm to 1840nm. The optical power on the sample was adjusted to measure the power-dependent fluorescence signal, and linear fitting of the fluorescence signal on a logarithmic scale was performed to determine the nonlinearity of the dye. Furthermore, this system can measure the wavelength-dependent ησ3, using 10μM Sulforhodamine 101 (SR101) as a reference. Thus, in this embodiment, the final concentration of the WGA-594 label was determined to be 5.6μM to ensure clear labeling.

[0075] In some optional embodiments of this example, after the step of acquiring the three-photon fluorescence image, the method further includes: injecting a WGA-594 marker into a second target location of the target object; when the reaction time of the WGA-594 marker is greater than a third preset time, obtaining a line graph showing the third marked area based on the third harmonic image and the three-photon fluorescence image.

[0076] Specifically, the third preset time can be 110 minutes, that is, when the reaction time of the WGA-594 label is greater than 110 minutes, the third harmonic image and the three-photon fluorescence image are merged to obtain the merged image (e.g., Figure 14 As shown in the image, a clear structure representing a smooth muscle cell layer can be seen on the outer side of the vessel wall labeled with WGA-594. This smooth muscle cell layer is located on the outer side of the lumen relative to the elastic fiber layer. In contrast, no such labeled cells are found on the vein wall imaged in the same field of view (e.g., Figures 15-17 As shown in the figure, the third marker region is used to represent the smooth muscle cell layer in the arterial wall structure.

[0077] It should be understood that the WGA-594 marker for VSMCs is not limited to small arteries; smooth muscle cell layers can also be seen in penetrating arteries. Figures 18-21 Two-dimensional (2D) images of penetrating arteries ranging from 132 to 168 micrometers below the surface of the brain are displayed. Individual smooth muscle cell layers on the penetrating artery walls are clearly shown, demonstrating that three-photon fluorescence imaging using WGA-594-labeled images can image smooth muscle cell layers on both arterioles and penetrating artery walls.

[0078] This embodiment also provides an imaging device for arterial wall structure, including a laser device, a filtering device, and a microscopic imaging system.

[0079] The laser device is used to emit laser signals with a wavelength of 1550nm;

[0080] The filtering device is used to preprocess the laser signal and send the generated soliton source signal to the microscopic imaging system;

[0081] The microscopic imaging system is used to scan and image the first target location of the target object, obtaining a third harmonic image and a three-photon fluorescence image; the third harmonic image and the three-photon fluorescence image are used to display the arterial wall structure of the target object.

[0082] The imaging device used for the aforementioned arterial wall structure, by pre-processing the laser signal to obtain a soliton source signal of a specific wavelength, exhibits better compatibility (stronger excitation characterization characteristics) with the WGA-594 label, thereby improving the imaging quality of the third harmonic imaging and three-photon fluorescence images after microscopic imaging. This achieves direct visualization and easy differentiation of different layers of the arterial wall structure. Furthermore, it demonstrates that the combination of third harmonic imaging and specific fluorescent labeling (WGA-594 label) is a powerful tool for imaging different layers and structures of the cerebral artery wall in vivo, and proves that WGA-594 can label the brain smooth muscle cell layer in vivo. Combined with third harmonic imaging, the endothelial layer, internal elastic layer, and smooth muscle cell layer of the cerebral artery wall in vivo can be distinguished.

[0083] It should be noted that, for the sake of simplicity, the foregoing method embodiments are all described as a series of actions. However, those skilled in the art should understand that this application is not limited to the described order of actions, as some steps may be performed in other orders or simultaneously according to this application. Furthermore, those skilled in the art should also understand that the embodiments described in the specification are preferred embodiments, and the actions and modules involved are not necessarily essential to this application.

[0084] In the above embodiments, the descriptions of each embodiment have different focuses. For parts not described in detail in a certain embodiment, please refer to the relevant descriptions in other embodiments.

[0085] The above are merely preferred embodiments of the present invention and do not limit the scope of the patent. Any equivalent structural or procedural transformations made based on the description and drawings of the present invention, or direct or indirect applications in other related technical fields, are similarly included within the scope of patent protection of the present invention.

Claims

1. An imaging method for arterial wall structure, characterized in that, include: The laser signal is preprocessed to obtain a soliton source signal; wherein the wavelength of the soliton source signal is 1665nm. The soliton light source signal is transmitted to the microscopic imaging system; The microscopic imaging system is used to perform imaging scanning on a first target position of the target object; wherein, the microscopic imaging system is equipped with a gallium arsenide photomultiplier tube and a gallium arsenide phosphide photomultiplier tube, and the first target position includes the head position; The third harmonic image is obtained using the gallium arsenide photomultiplier tube; An Alexa fluor633 marker is injected into a second target location of the target object, and a three-photon fluorescence image is acquired using the gallium arsenide phosphide photomultiplier tube; wherein, the third harmonic image and the three-photon fluorescence image are used to display the arterial wall structure of the target object, and the second target location includes the eye socket; When the reaction time of the Alexa fluor 633 marker is greater than a first preset time, the third harmonic image and the three-photon fluorescence image are merged to obtain a line graph showing a first marked region; wherein, the first marked region is used to represent the elastic fiber layer in the arterial wall structure; Inject WGA-594 markers at the second target location of the target object; When the reaction time of the WGA-594 marker is greater than the second preset time, the third harmonic image and the three-photon fluorescence image are merged to obtain a line graph showing the second marker region; wherein, the second marker region is used to represent vascular endothelial cells in the arterial wall structure, and the second preset time is greater than the first preset time; When the reaction time of the WGA-594 marker is greater than a third preset time, the third harmonic image and the three-photon fluorescence image are merged to obtain a line graph showing a third marker region; wherein, the third marker region is used to represent the smooth muscle cell layer in the arterial wall structure.

2. The imaging method for arterial wall structure as described in claim 1, characterized in that, Prior to the step of preprocessing the laser signal, the method further includes: Start the laser device and connect it to the filter device; The laser device emits a laser signal with a wavelength of 1550 nm to the filtering device.

3. The imaging method for arterial wall structure as described in claim 2, characterized in that, The step of preprocessing the laser signal to obtain the soliton source signal specifically includes: The laser signal is filtered by a filtering device to generate a soliton light source signal with a wavelength of 1665nm.

4. The imaging method for arterial wall structure as described in claim 3, characterized in that, The step of transmitting the soliton light source signal to the microscopic imaging system specifically includes: The soliton light source signal is guided by a signal guiding device to be emitted to a microscopic imaging system; wherein, the signal guiding device includes a plurality of lens structures for refracting the soliton light source signal.

5. The imaging method for arterial wall structure as described in claim 1, characterized in that, The gallium arsenide photomultiplier tube is equipped with a 540 / 80 nm bandpass filter, and the gallium arsenide phosphide photomultiplier tube is equipped with a 630 / 82 nm bandpass filter.

6. An imaging device for arterial wall structure, characterized in that, It includes a laser device, a filtering device, and a microscopic imaging system, wherein the microscopic imaging system is equipped with a gallium arsenide photomultiplier tube and a gallium arsenide phosphide photomultiplier tube; The laser device is used to emit laser signals with a wavelength of 1550 nm; The filtering device is used to preprocess the laser signal and send the generated soliton source signal to the microscopic imaging system; The microscopic imaging system is used to perform imaging scanning on a first target position of a target object, acquire a third harmonic image through a gallium arsenide photomultiplier tube, inject an Alexa fluor 633 marker into a second target position of the target object, acquire a three-photon fluorescence image through a gallium arsenide phosphorus photomultiplier tube, and when the reaction time of the Alexa fluor 633 marker is greater than a first preset time, obtain a line graph showing the first marked area based on the third harmonic image and the three-photon fluorescence image. A WGA-594 marker is injected into a second target location of the target object. When the reaction time of the WGA-594 marker is greater than a second preset time, a line graph showing the second marked region is obtained based on the third harmonic image and the three-photon fluorescence image. When the reaction time of the WGA-594 marker is greater than a third preset time, a line graph showing the third marked region is obtained based on the third harmonic image and the three-photon fluorescence image. The third harmonic image and the three-photon fluorescence image are used to display the arterial wall structure of the target object. The first marked region is used to represent the elastic fiber layer in the arterial wall structure. The second marked region is used to represent the vascular endothelial cells in the arterial wall structure. The third marked region is used to represent the smooth muscle cell layer in the arterial wall structure.

7. The imaging device for arterial wall structure as described in claim 6, characterized in that, The microscopic imaging system is equipped with a gallium arsenide photomultiplier tube with a 540 / 80 nm bandpass filter and a gallium arsenide phosphide photomultiplier tube with a 630 / 82 nm bandpass filter. The gallium arsenide photomultiplier tube is used to acquire the third harmonic image corresponding to the target object, and the gallium arsenide phosphorus photomultiplier tube is used to acquire the three-photon fluorescence image corresponding to the target object.

Images