Neurovascular-based coupled imaging systems, methods, devices, and storage media

Abstract

The application provides a neurovascular coupling imaging system, method, device and storage medium, which comprises an image collector, a microscope frame arranged in the light inlet direction of the image collector, a visible light system for emitting visible light to a sample to be collected after the visible light is reflected by a beam splitter in the microscope frame, a near-infrared light system for emitting near-infrared light to the sample to be collected after the near-infrared light is reflected by another beam splitter in the microscope frame, and a trigger device for switching the frequency of the time-sharing emission of each light source in the visible light system and the near-infrared light system based on the collection frequency of the image collector. The application can integrate different visible light or near-infrared laser light sources in the light path to be collected, is compatible with various imaging requirements such as visible light for imaging of nerve cell activity and near-infrared light speckle imaging for non-invasive blood flow imaging, ensures that there is no time difference between image collection and light source switching and no time resolution is lost, and facilitates subsequent storage and calculation of data.

Description

Technical Field

[0001] This invention relates to the field of microscopic imaging, and more specifically, to a neurovascular-based coupled imaging system, method, device, and storage medium. Background Technology

[0002] The nervous system is composed of various components, including nerve cells, glial cells, and blood vessels. Neurovascular coupling is an important area of ​​neuroscience research, involving the interaction between neural activity and vascular responses. This coupling mechanism is crucial for understanding brain function and its manifestations under various physiological and pathological conditions. In recent years, with the advancement of imaging technology, researchers have been able to observe and analyze the dynamic interactions between nerve cells and blood vessels more precisely. However, due to the sensitive and complex nature of the nervous system's response to external environmental stimuli, some technical and methodological limitations still exist.

[0003] Existing neurovascular coupling imaging techniques, such as functional magnetic resonance imaging (fMRI) and functional ultrasound imaging (fUS), typically rely on indirect dynamic changes in blood flow rather than direct measurements of nerve cell activity, providing some information about the relationship between blood flow changes and neural activity. However, studies have shown that cellular activity and extracellular activity not only possess their own spatiotemporal characteristics under different physiological and pathological states, but they also exhibit different dynamic interaction characteristics under different states. For example, good neurovascular coupling under physiological conditions can be lost under pathological conditions of severe ischemia and hypoxia. Because non-invasive in vivo animal imaging using functional ultrasound and brain MRI characterizes nerve cell activity using vascular activity rather than true nerve cell activity, the true spatiotemporal characteristics of the nervous system cannot be recorded under these neurovascular decoupling conditions.

[0004] Furthermore, although ultrasound functional brain imaging and brain magnetic resonance imaging (MRI) can utilize the characteristics of neurovascular coupling to conduct large-scale neurological studies, these techniques often struggle to simultaneously meet high-standard research requirements in terms of both temporal and spatial resolution. For example, while ultrasound imaging offers good temporal resolution, its spatial resolution and imaging depth are limited; and while MRI provides high spatial resolution, its temporal resolution and cost-effectiveness are limiting factors.

[0005] More importantly, existing technologies often struggle to conduct long-term monitoring without burdening the organism. Given the immune environment independence of the nervous system and its complex responses to external stimuli, avoiding imaging aids or imaging properties such as injection, contact, and noise is crucial when studying the interaction between cellular and extracellular activities in the nervous system of live animals. Many imaging techniques require invasive markers or specific environmental conditions, which can interfere with normal physiological states and affect the accuracy and reliability of experimental results.

[0006] Currently, there is no equipment that can non-invasively and simultaneously use neural cell fluorescence imaging and laser speckle imaging to image extracellular activities such as intracellular cell activity and vascular flow in the nervous system with a large field of view and high spatiotemporal resolution. Research on intracellular and extracellular interactions at the neural network level, especially neurovascular interactions, urgently needs new equipment support.

[0007] Therefore, the present invention provides a neurovascular coupling imaging system, method, device and storage medium. Summary of the Invention

[0008] To address the problems in the prior art, the present invention aims to provide a neurovascular-based coupled imaging system, method, device, and storage medium, which overcomes the difficulties of the prior art. By integrating different visible light or near-infrared laser light sources into the optical path to be acquired, it is compatible with various imaging requirements such as visible light for nerve cell activity and near-infrared speckle imaging for non-invasive blood flow imaging. It ensures that there is no time difference between image acquisition and light source switching without loss of time resolution, and facilitates subsequent data storage and calculation.

[0009] Embodiments of the present invention provide a neurovascular coupling imaging method, comprising the following steps:

[0010] An image acquisition device;

[0011] A microscope frame is positioned in the light-incoming direction of the image acquisition device;

[0012] A visible light system that reflects visible light through a beam splitter in the microscope frame and then emits it onto the sample.

[0013] A near-infrared light system emits near-infrared light onto the sample after it is reflected by another beam splitter in the microscope frame; and

[0014] A triggering device switches the time-division emission frequencies of each light source in the visible light system and near-infrared light system based on the acquisition frequency of the image acquisition device.

[0015] Preferably, the acquisition frequency of the image acquisition device includes a first high level and a second high level that are spaced apart from each other. The high level of the visible light system that triggers the emission in the operating frequency overlaps with the first high level in time. The high level of the near-infrared light system that triggers the emission in the operating frequency overlaps with the second high level in time. A visible light image set is established based on the first image acquired within the first high level, and a near-infrared light image set is established based on the second image acquired within the second high level.

[0016] Preferably, the system further includes a data processing module connected to the image acquisition unit. This module obtains a first set of high-level signals in time sequence based on the high-level signals of several different wavelengths of visible light emitted time-division multiple times within the operating frequency of the visible light system. It also obtains a second set of high-level signals in time sequence based on the high-level signals of at least one different wavelength of near-infrared light emitted time-division multiple times within the operating frequency of the near-infrared light system. A third set of high-level signals is obtained by superimposing the first and second sets of high-level signals in time sequence. The image acquisition unit performs image acquisition based on the third set of high-level signals and classifies and stores the images in the data processing module according to the mapping relationship between different wavelengths and the high-level signal timing.

[0017] Preferably, the microscope frame is disposed between the image acquisition device and the observation carrier, and the microscope frame includes a filter, a first beam splitter, a second beam splitter, and an objective lens arranged sequentially along the optical path;

[0018] The visible light system is an LED light source system that emits visible light of a preset wavelength towards the first beam splitter, and the light is reflected by the first beam splitter and then emitted onto the object to be sampled.

[0019] The near-infrared light system is a near-infrared laser source system that emits near-infrared laser light into the second beam splitter, and the laser light is reflected by the second beam splitter and then emitted onto the sample to be sampled.

[0020] The triggering device is connected to the image acquisition unit, the LED light source system, and the near-infrared laser light source system respectively. The triggering device switches the first frequency of each light source in the LED light source system and the near-infrared laser light source system to emit light in a time-division manner, which matches the second frequency at which the triggering device drives the image acquisition unit to acquire images.

[0021] Preferably, the LED light source system is equipped with at least one narrowband visible light LED light source selected from the 405nm, 488nm, 535nm, and 647nm bands; the first beam splitter is a non-polarized wideband planar beam splitter, which forms a 45° angle with the incident light formed by the beam emitted by the LED light source, and the reflected light forms a 45° angle with the first beam splitter and is perpendicular to the plane of the observation carrier carrying the sample. The beam splitting band of the first beam splitter is in the range of 400 to 800nm, and the ratio of the transmittance T to the reflectance R is 1:1.

[0022] Preferably, the second beam splitter is a non-polarized wide-band planar beam splitter, which forms a 45° angle with the incident light formed by the beam emitted by the near-infrared laser source. The reflected light forms a 45° angle with the second beam splitter and is perpendicular to the plane of the observation carrier carrying the sample. The beam splitting band of the second beam splitter is 400 to 800 nm, and the ratio of the transmittance T to the reflectance R is 1:1.

[0023] Preferably, the LED light source system and the near-infrared laser light source system are located on opposite sides of the microscope frame, the plane of the first beam splitter is perpendicular to the plane of the second beam splitter, the objective lens is a combination of microscope plates with variable magnification and adjustable focal length, the observation carrier carries nerve and blood vessel tissue, and the optical axis of the reflected light from the first beam splitter and the optical axis of the reflected light from the second beam splitter are coaxial with the optical axis of the incident light from the image acquisition device.

[0024] Preferably, the high level in the operating timing pulse of the LED light source system 1 is the emission state, the high level in the operating timing pulse of the near-infrared laser light source system 2 is the emission state, and the high level in the operating timing pulse of the image acquisition device is the acquisition state. The falling edge of the high level in the operating timing pulse of the LED light source system 1 and the rising edge of the high level in the operating timing pulse of the near-infrared laser light source system 2 overlap in timing, and this overlapping timing overlaps in timing with the low level between adjacent high levels in the operating timing pulse of the image acquisition device (or, the switching timing between the LED light source system 1 and the near-infrared laser light source system 2 is included in the timing of the low level, and the light source switching only occurs within the low level timing), thereby greatly reducing the low level in the image acquisition device and reducing the impact of switching different light sources on the continuity of observation.

[0025] Preferably, the combination of each low level between adjacent high levels in the operating timing pulse of the image acquisition device and the rising and falling edges adjacent to each low level is used as the non-acquisition timing (where each cycle consists of three parts: a falling edge of a high level + a low level + a rising edge of the next high level). The switching timing between the LED light source system 1 and the near-infrared laser light source system 2 occurs only within the non-acquisition timing. The switching timing is the combination of the falling edge of the high level in the operating timing pulse of the LED light source system 1 and the rising edge of the high level in the operating timing pulse of the near-infrared laser light source system 2.

[0026] Embodiments of the present invention also provide a neurovascular-based coupling imaging method, employing the aforementioned neurovascular-based coupling imaging device, comprising:

[0027] S110. Based on the high level of at least one different band of visible light emitted in time-division multiplexing at the operating frequency of the visible light system, a first set of high levels in time sequence is established.

[0028] S120. Based on the high level of at least one different band of near-infrared light emitted in time division at the operating frequency of the near-infrared light system, a second set of high levels is established in time sequence.

[0029] S130. Obtain the third high-level set by superimposing the first high-level set and the second high-level set in timing.

[0030] S140, The image acquisition device continuously acquires images based on the third high-level set; and

[0031] S150. Obtain a visible light image sequence based on the first image acquired during the first high level, and obtain a near-infrared light image sequence based on the second image acquired during the second high level.

[0032] Embodiments of the present invention also provide a neurovascular coupling imaging device, comprising:

[0033] processor;

[0034] A memory in which executable instructions of the processor are stored;

[0035] The processor is configured to perform the steps of the above-described neurovascular-based coupled imaging method by executing the executable instructions.

[0036] Embodiments of the present invention also provide a computer-readable storage medium for storing a program that, when executed, implements the steps of the above-described neurovascular-based coupled imaging method.

[0037] The purpose of this invention is to provide a neurovascular-based coupled imaging system, method, device, and storage medium that can integrate different visible light or near-infrared laser light sources into the optical path to be acquired, thus being compatible with various imaging requirements such as visible light for nerve cell activity and near-infrared speckle imaging for non-invasive blood flow imaging. This ensures that there is no time difference between image acquisition and light source switching without loss of temporal resolution, and facilitates subsequent data storage and calculation. Attached Figure Description

[0038] Other features, objects, and advantages of the invention will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings.

[0039] Figure 1 This is a schematic diagram of the modules of the neurovascular coupling imaging system of the present invention.

[0040] Figure 2 This is a schematic diagram of the appearance of the neurovascular-based coupled imaging system of the present invention.

[0041] Figure 3 This is a schematic diagram of the optical path in the neurovascular-based coupled imaging system of the present invention.

[0042] Figure 4 This is a schematic diagram illustrating the principle of obtaining different images using the neurovascular-based coupled imaging system of this invention.

[0043] Figure 5 This is a schematic flowchart of the neurovascular coupling imaging method of the present invention.

[0044] Figure 6 This is a schematic diagram of the structure of the neurovascular coupling imaging device of the present invention.

[0045] Figure 7 This is a schematic diagram of the structure of a computer-readable storage medium according to an embodiment of the present invention.

[0046] Figure Labels

[0047] 1 LED Light Source System

[0048] 2. Near-infrared laser source system

[0049] 3. Triggering device

[0050] 4. Microscope stand

[0051] 401 filter

[0052] 402 First Beam Spectroscope

[0053] 403 Second Beam Spectroscope

[0054] 404 objective lens

[0055] 5 Image Acquisition Device

[0056] 6. Data Processing Module Detailed Implementation

[0057] The following specific examples illustrate the implementation methods of this application. Those skilled in the art can easily understand the other advantages and effects of this application from the content disclosed herein. This application can also be implemented or applied through other different specific embodiments, and various details in this application can be modified or changed according to different viewpoints and application systems without departing from the spirit of this application. It should be noted that, unless otherwise specified, the embodiments and features in the embodiments of this application can be combined with each other.

[0058] The embodiments of this application will now be described in detail with reference to the accompanying drawings, so that those skilled in the art can easily implement the application. This application may be embodied in many different forms and is not limited to the embodiments described herein.

[0059] In this application, the terms "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics represented in connection with that embodiment or example, which are included in at least one embodiment or example of this application. Furthermore, the specific features, structures, materials, or characteristics represented may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate different embodiments or examples represented in this application, as well as features of different embodiments or examples.

[0060] Furthermore, the terms "first" and "second" are used for illustrative purposes only and should not be construed as indicating or implying 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 representation of this application, "multiple" means two or more, unless otherwise explicitly specified.

[0061] For the purpose of clearly describing this application, devices that are not relevant to the description are omitted, and the same or similar components throughout the specification are given the same reference numerals.

[0062] Throughout this specification, when it is said that a device is "connected" to another device, this includes not only "direct connection" but also "indirect connection" by placing other components in between. Furthermore, when it is said that a device "comprises" a certain constituent element, unless otherwise stated otherwise, this does not exclude other constituent elements, but rather implies that other constituent elements may be included.

[0063] When we say that a device is "above" another device, this can mean that it is directly above the other device, or it can mean that other devices are present in between. Conversely, when we say that a device is "directly" "above" another device, there are no other devices present in between.

[0064] Although the terms first, second, etc., are used in some instances herein to refer to various elements, these elements should not be limited by these terms. These terms are used only to distinguish one element from another. For example, first interface and second interface, etc., are used. Furthermore, as used herein, the singular forms “a,” “an,” and “the” are intended to also include the plural forms unless the context indicates otherwise. It should be further understood that the terms “comprising,” “including,” indicate the presence of features, steps, operations, elements, components, items, kinds, and / or groups, but do not exclude the presence, occurrence, or addition of one or more other features, steps, operations, elements, components, items, kinds, and / or groups. The terms “or” and “and / or” as used herein are interpreted as inclusive, or mean any one or any combination thereof. Thus, “A, B, or C” or “A, B, and / or C” means “any one of: A; B; C; A and B; A and C; B and C; A, B, and C.” Exceptions to this definition will only occur if the combination of elements, functions, steps, or operations is inherently mutually exclusive in some way.

[0065] The technical terms used herein are for reference only to specific embodiments and are not intended to limit the scope of this application. The singular form used herein includes the plural form unless the statement explicitly indicates otherwise. The word "comprising" as used in the specification means to specify a particular characteristic, region, integer, step, operation, element, and / or component, and does not exclude the presence or addition of other characteristics, regions, integers, steps, operations, elements, and / or components.

[0066] Although not explicitly defined, all terms, including technical and scientific terms used herein, shall have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. Terms defined in commonly used dictionaries shall be further interpreted as having a meaning consistent with the relevant technical literature and the content of this present application, and shall not be over-interpreted as having an ideal or overly formulaic meaning unless otherwise defined.

[0067] Currently, neural cell fluorescence imaging technology, by exciting fluorescent proteins in specific cell types in transgenic mice with a light source of a specific wavelength, can directly observe and record specific cellular activities. This technology is widely used in experimental research in neuroscience because it can directly reflect the physiological state of cells. Meanwhile, laser speckle imaging technology, by emitting a coherent light source and analyzing changes in the intensity of scattered light, can measure blood flow dynamics in blood vessels in real time. This method is not only non-invasive and non-surgical, but also provides high temporal resolution, making it suitable for long-term continuous monitoring.

[0068] Figure 1 This is a schematic diagram of the modules of the neurovascular coupling imaging system of the present invention. Figure 2 This is a schematic diagram of the appearance of the neurovascular-based coupled imaging system of the present invention. Figure 3 This is a schematic diagram of the optical path in the neurovascular-based coupled imaging system of the present invention. Figure 4 This is a schematic diagram illustrating the principle of obtaining different images using the neurovascular-based coupled imaging system of this invention. To facilitate the distinction between the optical path and the physical circuitry, Figure 1 , 3 In section 4, the dashed arrow A represents the light path after emission or reflection from the light source, and the straight arrow B represents the data transmission line. For example... Figures 1 to 4 As shown, this invention provides a neurovascular coupling imaging device, specifically a large-field-of-view neurovascular coupling brain mesoscopic functional imaging device combining visible and near-infrared light, comprising: a visible light system, a near-infrared light system, a triggering device 3, a microscope gantry 4, and an image acquisition unit 5. The microscope gantry 4 is positioned in the light-incoming direction of the image acquisition unit 5. The visible light system reflects visible light through a beam splitter in the microscope gantry 4 and then emits it onto the sample. The near-infrared light system reflects near-infrared light through another beam splitter in the microscope gantry 4 and then emits it onto the sample. The triggering device 3 switches the time-division emission frequencies of the light sources in the visible light system and the near-infrared light system based on the acquisition frequency of the image acquisition unit 5. This invention integrates different visible light or near-infrared laser light sources into the optical path to be acquired, accommodating various imaging needs such as visible light for neural cell activity and near-infrared speckle imaging for non-invasive blood flow imaging. It ensures that there is no time lag between image acquisition and light source switching without sacrificing temporal resolution, and facilitates subsequent data storage and computation.

[0069] In a preferred embodiment, the acquisition frequency of the image acquisition device 5 includes a first high level and a second high level that are spaced apart from each other. The high level that triggers the emission in the operating frequency of the visible light system overlaps with the first high level in timing, and the high level that triggers the emission in the operating frequency of the near-infrared light system overlaps with the second high level in timing. A visible light image set is established based on the first image acquired within the first high level, and a near-infrared light image set is established based on the second image acquired within the second high level, but this is not a limitation.

[0070] In a preferred embodiment, a data processing module 6 is further included, connected to the image acquisition unit 5. Based on the high levels of several different wavelengths of visible light emitted in a time-division multiplexing manner within the operating frequency of the visible light system, a first high-level set in time sequence is obtained. Based on the high levels of at least one different wavelength of near-infrared light emitted in a time-division multiplexing manner within the operating frequency of the near-infrared light system, a second high-level set in time sequence is obtained. A third high-level set is obtained by superimposing the first and second high-level sets in time sequence. The image acquisition unit 5 performs image acquisition based on the third high-level set and classifies and stores the images in the data processing module 6 according to the mapping relationship between different wavelengths and high-level time sequences, but this is not a limitation.

[0071] In a preferred embodiment, a microscope gantry 4 is positioned between the image acquisition device 5 and the observation carrier. The microscope gantry 4 includes a filter 401, a first beam splitter 402, a second beam splitter 403, and an objective lens 404 arranged sequentially along the optical path. The visible light system is an LED light source system 1, which emits visible light of a preset wavelength towards the first beam splitter 402, and the light is reflected by the first beam splitter 402 before being emitted onto the object to be sampled. The near-infrared light system is a near-infrared laser light source system 2, which emits near-infrared laser light towards the second beam splitter 403, and the laser is reflected by the second beam splitter 403 before being emitted onto the object to be sampled. A triggering device 3 is connected to the image acquisition device 5, the LED light source system 1, and the near-infrared laser light source system 2, respectively. The triggering device 3 switches the first frequency emitted by each light source in the LED light source system 1 and the near-infrared laser light source system 2 to match the second frequency at which the triggering device 3 drives the image acquisition device 5 to acquire images, but this is not a limitation.

[0072] In a preferred embodiment, the LED light source system 1 is equipped with at least one narrowband visible light LED source selected from the 405nm, 488nm, 535nm, and 647nm bands. The first beam splitter 402 is a non-polarized wideband planar beam splitter, which forms a 45° angle with the incident light formed by the beam emitted by the LED light source. The reflected light forms a 45° angle with the first beam splitter 402 and is perpendicular to the plane of the observation carrier carrying the sample. The beam splitting band of the first beam splitter 402 is in the range of 400 to 800 nm, and the ratio of the transmittance T to the reflectance R is 1:1, but not limited thereto.

[0073] In a preferred embodiment, the second beam splitter 403 is a non-polarized broadband planar beam splitter that forms a 45° angle with the incident light formed by the beam emitted by the near-infrared laser source. The reflected light forms a 45° angle with the second beam splitter 403 and is perpendicular to the plane on which the observation carrier carrying the sample is located. The beam splitting band of the second beam splitter 403 is in the range of 400 to 800 nm, and the ratio of the transmittance T to the reflectance R is 1:1, but is not limited thereto.

[0074] In a preferred embodiment, the LED light source system 1 and the near-infrared laser light source system 2 are located on opposite sides of the microscope frame 4, thereby allowing the LED light source system 1 and the near-infrared laser light source system 2 to have more space and helping to reduce the overall height of the neurovascular-based coupled imaging system. The plane where the first beam splitter 402 is located is perpendicular to the plane where the second beam splitter 403 is located. The objective lens 404 is a combination of microscope plates with variable magnification and adjustable focal length. The observation carrier carries neurovascular tissue. The optical axes of the reflected light from the first beam splitter 402 and the reflected light from the second beam splitter 403 are coaxial with the optical axis of the incident light from the image acquisition device 5, but this is not a limitation.

[0075] In a preferred embodiment, the high level in the operating timing pulse of the LED light source system 1 is the emission state, the high level in the operating timing pulse of the near-infrared laser light source system 2 is the emission state, and the high level in the operating timing pulse of the image acquisition device is the acquisition state. The falling edge of the high level in the operating timing pulse of the LED light source system 1 and the rising edge of the high level in the operating timing pulse of the near-infrared laser light source system 2 overlap in timing, and this overlapping timing overlaps in timing with the low level between adjacent high levels in the operating timing pulse of the image acquisition device. This can greatly reduce the low level in the image acquisition device and reduce the impact of switching different light sources on the continuity of observation, but it is not limited thereto.

[0076] This invention aims to address a series of technical problems and limitations in existing neurovascular coupling imaging techniques, in order to provide more accurate and comprehensive measurements of neural activity and its synchronous measurement with vascular dynamics. Specifically, this invention focuses on solving the following key technical problems:

[0077] ① Limitations of indirect imaging: Existing neurovascular coupling imaging methods, such as functional magnetic resonance imaging (fMRI) and near-infrared spectroscopy (NIRS), largely rely on changes in blood flow to indirectly infer neural activity, making it impossible to directly observe the precise activity of nerve cells. This indirect measurement method may miss crucial physiological information, especially in pathological states of neurovascular decoupling.

[0078] ② Limitations of temporal and spatial resolution: Although existing technologies can provide a certain degree of temporal or spatial resolution, they often cannot simultaneously meet the requirements for high temporal and high spatial resolution. For example, ultrasound imaging has good temporal resolution, but its spatial resolution and imaging depth are limited, while magnetic resonance imaging is insufficient in terms of temporal resolution.

[0079] ③ Challenges of long-term continuous monitoring: Current imaging technologies often cannot perform long-term continuous monitoring without introducing biological burden. Many imaging technologies require invasive labeling or special environmental conditions, which may affect the normal physiological state of organisms and the accuracy of experimental results.

[0080] ④ High cost and operational complexity: Many advanced imaging devices, such as MRI machines, are not only expensive but also complex to operate, requiring professional personnel for operation and maintenance, which limits their widespread use in ordinary laboratory and clinical settings.

[0081] This invention provides a non-invasive imaging device with a wide field of view and high spatiotemporal resolution that can simultaneously perform cellular and extracellular activities in the nervous system. Specifically, it is a device that uses a visible light narrow-band LED light source and a near-infrared laser light source to emit light to the same imaging area through optical path design, and uses a controllable triggering device to align the timing of image capture, light source emission, and light source switching, thereby achieving a wide field of view and high spatiotemporal resolution optical imaging and image acquisition of the area after different light source emission.

[0082] The present invention achieves its objectives of multi-source, wide-field-of-view, and high spatiotemporal resolution optical imaging and image acquisition of the sample (usually a live animal) primarily through the following device design:

[0083] Continue to refer to Figures 1 to 4 This invention relates to a large-field-of-view microscopic imaging and high spatiotemporal resolution acquisition device for nerve blood flow, comprising an LED light source system 1, a near-infrared laser light source system 2, a triggering device 3, a microscope frame 4, a filter 401, a first beam splitter 402, a second beam splitter 403, an objective lens 404, an image acquisition device 5, and a data processing module 6. The data processing module 6 is used for data acquisition, display, and storage.

[0084] The LED light source system 1 and the near-infrared laser light source system 2 are respectively positioned on opposite sides of the microscope frame 4. The LED light source system 1 emits visible light of a preset wavelength towards the first beam splitter 402, and the light is reflected by the first beam splitter 402 before being emitted onto the sample. The near-infrared laser light source system 2 emits near-infrared laser light towards the second beam splitter 403, and the laser is reflected by the second beam splitter 403 before being emitted onto the sample. The plane containing the first beam splitter 402 is perpendicular to the plane containing the second beam splitter 403.

[0085] The LED light source system 1 can simultaneously carry narrowband visible light source channels in the 405nm, 488nm, 535nm, and 647nm wavelength bands, and the light intensity can be adjusted. The LED light source system is mechanically assembled and connected to the microscope frame, and the emitted light source is parallel to the plane of the sample. Each light source channel is connected to the triggering device through a BNC interface.

[0086] The near-infrared laser source system 2 mainly emits 780nm narrowband near-infrared laser light, and the light intensity can be adjusted. The near-infrared laser source is mechanically assembled and connected to the microscope frame, and the source channel is also connected to the triggering device through a BNC interface.

[0087] The microscope frame 4 is perpendicular to the plane of the sample. Inside it, a filter, a first beam splitter, a second beam splitter, and an objective lens are loaded in sequence from the far end to the near end of the sample. The optical axis of the filter, the first beam splitter, the second beam splitter, and the objective lens is coaxial and projected onto the center of the sample.

[0088] Filter 401 is a multi-channel filter that can block narrowband visible light of 405nm, 488nm, 535nm, 647nm, etc. emitted by the LED light source system, as well as other visible light of other bands emitted by the sampled object after being emitted by the LED light source system and near-infrared light emitted by the near-infrared laser source and reflected by the sampled object.

[0089] The first beam splitter 402 is a non-polarized wide-band planar beam splitter with a transmittance T: reflectance R ratio of 1:1. The beam splitting band includes 400-800nm. It forms a 45° angle with the incident light formed by the beam emitted by the LED light source. The reflected light forms a 45° angle with the first beam splitter and is perpendicular to the sample.

[0090] The second beam splitter 403 is a non-polarized wide-band planar beam splitter with a transmittance:reflectance (T:R) ratio of 1:1. The beam splitting band includes 400-800nm. It forms a 45° angle with the incident light formed by the beam emitted by the near-infrared laser source. The reflected light forms a 45° angle with the second beam splitter and is perpendicular to the sample.

[0091] Objective lens 404 is a variable magnification microscope slide assembly that allows for focal length adjustment, enabling adjustment of the image size of the object under test in the image acquisition device.

[0092] The triggering device 3 is connected to the image acquisition unit, one or more light source channels in the LED light source system, and the near-infrared laser light source channel via multiple BNC interfaces. The triggering device receives the image acquisition frequency parameters from the image acquisition unit and aligns the frequency to each light source channel and the near-infrared laser light source channel in the LED light source system. While the image acquisition unit acquires each image, it switches between the light sources to form a high-frequency image acquired by the image acquisition unit after each light source channel alternately emits the sampled object.

[0093] Image acquisition unit 5 is a high-speed, high-resolution back-illuminated CMOS camera with a pixel count sufficient for large-field imaging of a 1.5 × 1.5 cm sample. The image acquisition unit is connected to the microscope frame, receiving images of the sample from the LED light source system and / or near-infrared laser light source system via the lens group (including objective lens, first beam splitter, beam splitter, and filters) within the microscope frame, and converting the optical signal to a digital signal. The image acquisition unit is also connected to the data processing module, receiving image acquisition parameters and commands from it and performing image acquisition based on these parameters and commands. Finally, the image acquisition unit is connected to a triggering device, sending the image acquisition frequency parameters to the triggering device via a BNC interface. In a preferred embodiment, the acquisition frequency of the image acquisition device 5 includes a first high level and a second high level that are spaced apart from each other. The high level that triggers the emission in the operating frequency of the visible light system overlaps with the first high level in timing. The high level that triggers the emission in the operating frequency of the near-infrared light system overlaps with the second high level in timing. A visible light image set is established based on the first image acquired within the first high level, and a near-infrared light image set is established based on the second image acquired within the second high level.

[0094] The data processing module 6 comprises a calculator and a display. Both the calculator and the display are connected to the image acquisition unit. The display can show the images acquired by the image acquisition unit in real time. The calculator can be used to set the acquisition parameters of the image acquisition unit and issue acquisition commands to activate the image acquisition unit for image acquisition. The calculator can also store the data acquired by the image acquisition unit. In a preferred embodiment, a first set of high-level signals in time sequence is obtained by time-division multiplexing several different bands of visible light within the operating frequency of the visible light system. A second set of high-level signals in time sequence is obtained by time-division multiplexing at least one different band of near-infrared light within the operating frequency of the near-infrared light system. A third set of high-level signals is obtained by superimposing the first and second sets of high-level signals in time sequence. The image acquisition unit 5 acquires images based on the third set of high-level signals and classifies and stores them in the data processing module 6 according to the mapping relationship between different bands and the high-level signal time sequence.

[0095] Compared with existing technologies, the present invention has the following advantages:

[0096] ① Multimodal imaging integration:

[0097] This invention, based on the properties of different visible or near-infrared laser light sources such as wavelength, bandwidth, and transmittance, and the respective imaging principles of visible light large-field fluorescence imaging and near-infrared laser speckle imaging, designs the optical path to integrate multiple bands of visible and near-infrared laser light into a single optical path perpendicular to the target image. This allows for compatibility with various imaging needs, such as visible light for neural cell activity and near-infrared speckle imaging for non-invasive blood flow imaging.

[0098] a. Neural cell fluorescence imaging module: Using gene editing technology, specific nerve cells express fluorescent proteins, are excited by a light source of a specific wavelength, and a high-speed camera records the fluorescence signal to reflect the activity state of the nerve cells.

[0099] b. Laser speckle imaging module: Uses a long-wavelength coherent light source to emit light non-contactly and non-invasively onto the tissue, records the light intensity image after diffuse reflection, and obtains the flow velocity information of the vascular system through image processing.

[0100] c. The present invention aligns the image acquisition frequency and the light source switching frequency through a trigger device within the device, so that there is no time difference between image acquisition and light source switching without losing time resolution, and facilitates subsequent data storage and calculation.

[0101] ②High spatiotemporal resolution:

[0102] a. Temporal resolution: The device is equipped with a high-speed camera and a fast light source switching system, which can capture instantaneous changes in neural activity and blood flow with a temporal resolution of milliseconds.

[0103] b. Spatial resolution: High-precision optical systems and image processing algorithms provide cellular-level spatial resolution, ensuring accurate imaging of neurons and tiny blood vessels.

[0104] ③ Wide field of view imaging:

[0105] The device has a wide field of view imaging capability, covering the entire brain region or a large area of ​​the cortex, and can simultaneously observe and compare the neural activity and vascular flow of multiple brain regions, and study the overall function of the brain network and the interaction of local regions.

[0106] ④ Non-invasive, non-contact imaging:

[0107] It employs non-invasive, non-contact imaging methods, avoiding physical interference and damage to brain tissue, and is suitable for long-term continuous observation, maintaining the animal's natural physiological state.

[0108] The technical, economic, and social benefits of this invention include:

[0109] ① Revealing the neurovascular coupling mechanism:

[0110] Neurovascular coupling refers to the interaction between neural activity and changes in blood flow, which is of great significance under both normal physiological and pathological conditions. This device can simultaneously record neural cell activity and blood flow, helping to elucidate their interaction mechanisms and promoting research on cerebrovascular diseases, neurodegenerative diseases, and other related conditions.

[0111] ②Supports long-term dynamic research:

[0112] Brain function and blood flow changes are dynamic processes, and long-term continuous observation is crucial for capturing their patterns and revealing complex physiological mechanisms. Non-invasive imaging methods enable this device to conduct long-term dynamic studies and obtain more comprehensive data.

[0113] ③ Study the overall function of brain networks:

[0114] The brain is a complex, integrated network, and the interactions between different brain regions are crucial to brain function. Wide-field imaging capabilities allow researchers to simultaneously observe the activity of multiple brain regions, study their collaborative mechanisms under different physiological and pathological conditions, and understand the overall function of the brain network.

[0115] ④ Reduce experimental interference:

[0116] Non-invasive imaging methods avoid additional interference to animals such as injections and contact, maintain the natural physiological state of experimental animals, help obtain more realistic data on neural activity and blood flow changes, reduce experimental errors, and improve the accuracy of research results.

[0117] ⑤ Advancing the forefront of neuroscience research:

[0118] This device fills a gap in existing technology and provides a powerful tool for neuroscience research. Its high spatiotemporal resolution, wide field of view, and non-invasive imaging capabilities can meet the urgent need of neuroscience research for simultaneous observation of nerve cell activity and blood flow, thus advancing the understanding and exploration of the complex mechanisms of the nervous system.

[0119] Figure 5 This is a schematic flowchart of the neurovascular coupling imaging method of the present invention. Figure 5 As shown, embodiments of the present invention also provide a neurovascular-based coupling imaging system for implementing the above-described neurovascular-based coupling imaging method, comprising the following steps:

[0120] S110. Based on the high level of at least one different band of visible light emitted in time division at the operating frequency of the visible light system, a first set of high levels is established in time sequence;

[0121] S120, Based on the high level of at least one different band of near-infrared light emitted in time division at the operating frequency of the near-infrared light system, a second set of high levels is established in time sequence;

[0122] S130. Obtain the third high-level set by superimposing the first high-level set and the second high-level set in timing.

[0123] S140, The image acquisition device continuously acquires images based on the third high-level set; and

[0124] S150. Obtain a visible light image sequence based on the first image acquired during the first high level, and obtain a near-infrared light image sequence based on the second image acquired during the second high level.

[0125] In a preferred embodiment, the high level in the operating timing pulse of the LED light source system 1 is the emission state, the high level in the operating timing pulse of the near-infrared laser light source system 2 is the emission state, and the high level in the operating timing pulse of the image acquisition device is the acquisition state. The triggering device 3 drives the LED light source system 1, the near-infrared laser light source system 2, and the image acquisition device 5, such that the falling edge of the high level in the operating timing pulse of the LED light source system 1 overlaps with the rising edge of the high level in the operating timing pulse of the near-infrared laser light source system 2 in timing. Furthermore, this overlapping timing overlaps with the low level between adjacent high levels in the operating timing pulse of the image acquisition device 5 in timing (i.e., fully utilizing the gap of the low level in the image acquisition device to achieve the switching of different light sources, ensuring that the high level period of the image acquisition device can be completely used for image acquisition). This can greatly reduce the low level in the image acquisition device and reduce the impact of switching different light sources on the continuity of observation, but it is not limited to this.

[0126] The neurovascular coupling imaging method of the present invention can integrate different visible light or near-infrared laser light sources into the optical path to be acquired, and is compatible with various imaging requirements such as visible light for nerve cell activity and near-infrared speckle imaging for non-invasive blood flow imaging. It ensures that there is no time difference between image acquisition and light source switching without loss of time resolution, and facilitates subsequent data storage and calculation.

[0127] The specific embodiments of the present invention are as follows:

[0128] The following is in conjunction with the appendix Figures 1 to 4 The present invention will be described in detail with reference to specific embodiments. These embodiments are based on the technical solutions of the present invention and provide detailed implementation methods and specific operating procedures. However, the scope of protection of the present invention is not limited to the following embodiments. Through this embodiment in conjunction with the appended drawings... Figures 1 to 4 The present invention provides a further description of its large-field-of-view microscopic imaging and high spatiotemporal resolution acquisition device for neural blood flow. This device can be used for large-field-of-view microscopic imaging and high spatiotemporal resolution acquisition of neural blood flow across the entire cortical region of a mouse brain.

[0129] Transgenic mice with transparent skulls, carrying fluorescent protein light intensity to characterize neural cell activity, were placed on the imaging plane, with their dorsal skulls positioned within the imaging field of view. Figure 1The imaging process diagram shows that this example utilizes an LED light source system 1, a near-infrared laser light source system 2, a trigger device 3, a microscope frame 4, a filter 401, a first beam splitter 402, a second beam splitter 403, an objective lens 404, an image acquisition unit 5, and a data processing module 6. The data processing module 6 can display the imaging results from the image acquisition unit 5 in real time. By adjusting either the LED light source system 1 or the near-infrared laser light source system 2 to emit light from the dorsal cortex of the mouse to be imaged, a real-time imaging image is obtained. Based on the imaging results of this image, the microscope frame 4 is adjusted for focusing, and the magnification of the objective lens is adjusted. The image acquisition area, acquisition frequency, number of alternating light source channels, light intensity, and image acquisition time are set via the computer in the data processing module 6.

[0130] After all parameters are determined, the image acquisition device 5 is triggered by the computer in the data processing module 6. The image acquisition device 5 then synchronously triggers the triggering device 3. The triggering device 3, according to the acquisition frequency of the image acquisition device, sequentially triggers the light source channels in the LED light source system 1 and / or the near-infrared laser light source system 2 that participate in imaging. Figure 4 As shown, the image acquisition device uses a triggering device to sequentially trigger multiple light source channels in each light source system to alternately supply light in sequence, thereby synchronizing image acquisition and light source emission. This reduces the impact of the timing gap between the rising and falling edges of different light source emission sequences on continuous observation (the falling edge of the high level in the working timing pulse of LED light source system 1 overlaps with the rising edge of the high level in the working timing pulse of near-infrared laser light source system 2, and this overlapping timing overlaps with the low level between adjacent high levels in the working timing pulse of the image acquisition device, thus greatly reducing the low level in the image acquisition device and reducing the impact of switching different light sources on the continuity of observation). The acquired data is synchronously stored in the computer, the data type is image data, and the naming order is the order of the acquired images.

[0131] Depend on Figure 3As shown in the optical path diagram, the light emitted by the LED light source system 1 is redirected by the first beam splitter 402 and emitted downwards through the second beam splitter 403 and objective lens 404 onto the dorsal skull of the mouse in the imaging field of view. Subsequently, the corresponding fluorescent protein carried by the transgenic mouse is excited by the specific wavelength of the LED light source emitted by the light source system, and the fluorescent protein emits light with a wavelength different from that of the LED light source. This light passes sequentially through objective lens 404, second beam splitter 403, first beam splitter 402, and multi-channel filter 401 before entering the image acquisition device for acquisition. The near-infrared laser light source system 2 emits a near-infrared laser light source, which is redirected by the second beam splitter 403 and emitted downwards through objective lens 404 onto the dorsal skull of the mouse in the imaging field of view. After diffuse reflection from the uneven skull surface, the near-infrared laser light passes sequentially through objective lens 404, second beam splitter 403, first beam splitter 402, and filter 401 before entering the image acquisition device 5 for acquisition. (In this embodiment, the LED light source system 1 has two visible light sources with different frequencies.) Figure 4 The image acquisition frequency of the image acquisition device is shown in two cycles formed by six high-level pulses. The first and fourth high levels correspond to the high level of the laser frequency of the first LED light source in LED light source system 1; the second and fifth high levels correspond to the high level of the laser frequency of the second LED light source in LED light source system 1; and the third and sixth high levels correspond to the laser frequency of the near-infrared laser in near-infrared laser light source system 2.

[0132] After image acquisition is complete, the image data stored in the computer needs to be processed, including light source channel identification, image smoothing, and other corrections. Since the acquired images are named according to the order of the light sources, the resulting...

[0133] The light source channel information used for a certain image = the remainder of the image name number / the number of light source channels involved in the acquisition;

[0134] The channel used to acquire the image is obtained accordingly. Then, all images in the same image acquisition sequence are classified according to their respective light source channels to obtain a high spatiotemporal resolution image sequence for each light source channel in a single image acquisition sequence. The temporal resolution of the image sequence acquired by each light source channel = image acquisition rate × number of light source channels participating in imaging.

[0135] The phase difference of each light source channel is fixed, for

[0136] Phase difference = image acquisition frequency × (number of intervals between two light sources + 1); further processing such as rigid correction and dynamic smoothing can be performed on the image. This image processing step can be done using commercial software such as MATLAB.

[0137] Figure 2This is a physical simulation diagram of the present invention, illustrating an LED light source system 1, a near-infrared laser light source system 2, a triggering device 3, a microscope frame 4, an image acquisition device 5, and their connection methods. The LED light source system 1, the near-infrared laser light source system 2, and the image acquisition device 5 are mechanically fixed to the microscope frame 4. This allows for the use of the applicant's other utility model invention, "A Fixation Device for Awake Imaging of Rodents," to fix awake mice for imaging, thus broadening the application scenarios.

[0138] This embodiment is merely for illustrating the specific principles and applicable scenarios, and is not limited thereto. For researchers in the art, subsequent modifications and variations based on this embodiment are also considered within the scope of protection of this embodiment.

[0139] This invention combines neural cell fluorescence imaging with laser speckle imaging technology to simultaneously record neural cell activity and vascular responses on the same imaging platform, offering significant advantages and unique application value. First, this simultaneous imaging method provides intuitive evidence of the direct interaction between neural activity and blood flow dynamics, contributing to a deeper understanding of the specific mechanisms of neurovascular coupling. Second, this combined technique allows for the simultaneous observation of changes in nerves and blood vessels under the same physiological or pathological conditions, enabling precise analysis of their dynamic relationship. This is particularly important for studying abnormal neurovascular interactions in conditions such as cerebral ischemia, hypoxia, and other neurodegenerative diseases.

[0140] This invention also provides a neurovascular-based coupling imaging device, including a processor and a memory storing executable instructions for the processor. The processor is configured to execute steps of a neurovascular-based coupling imaging method via the execution of the executable instructions.

[0141] As shown above, the neurovascular coupling imaging device of this invention in this embodiment can integrate different visible light or near-infrared laser light sources into the optical path to be acquired, and is compatible with various imaging requirements such as visible light for nerve cell activity and near-infrared speckle imaging for non-invasive blood flow imaging. It ensures that there is no time difference between image acquisition and light source switching without losing time resolution, and facilitates subsequent data storage and calculation.

[0142] Those skilled in the art will understand that various aspects of the present invention can be implemented as systems, methods, or program products. Therefore, various aspects of the present invention can be specifically implemented in the following forms: a completely hardware implementation, a completely software implementation (including firmware, microcode, etc.), or a combination of hardware and software aspects, collectively referred to herein as a "circuit," "module," or "platform."

[0143] Figure 6This is a schematic diagram of the neurovascular coupling imaging device of the present invention. See below for reference. Figure 6 To describe an electronic device 600 according to this embodiment of the present invention. Figure 6 The electronic device 600 shown is merely an example and should not impose any limitations on the functionality and scope of use of the embodiments of the present invention.

[0144] like Figure 6 As shown, the electronic device 600 is presented in the form of a general-purpose computing device. The components of the electronic device 600 may include, but are not limited to: at least one processing unit 610, at least one storage unit 620, a bus 630 connecting different platform components (including storage unit 620 and processing unit 610), a display unit 640, etc.

[0145] The storage unit stores program code, which can be executed by the processing unit 610 to perform the steps described in the method section of this specification according to various exemplary embodiments of the present invention. For example, the processing unit 610 can perform actions such as... Figure 1 The steps are shown in the figure.

[0146] Storage unit 620 may include a readable medium in the form of a volatile storage unit, such as random access memory (RAM) 6201 and / or cache memory 6202, and may further include a read-only memory (ROM) 6203.

[0147] Storage unit 620 may also include a program / utility 6204 having a set (at least one) program module 6205, such program module 6205 including but not limited to: operating system, one or more application programs, other program modules and program data, each or some combination of these examples may include an implementation of a network environment.

[0148] Bus 630 can represent one or more of several types of bus structures, including a memory cell bus or memory cell controller, a peripheral bus, a graphics acceleration port, a processing unit, or a local bus using any of the multiple bus structures.

[0149] Electronic device 600 can also communicate with one or more external devices 700 (e.g., keyboard, pointing device, Bluetooth device, etc.), and with one or more devices that enable a user to interact with electronic device 600, and / or with any device that enables electronic device 600 to communicate with one or more other computing devices (e.g., router, modem, etc.). This communication can be performed via input / output (I / O) interface 650. Furthermore, electronic device 600 can also communicate with one or more networks (e.g., local area network (LAN), wide area network (WAN), and / or public networks, such as the Internet) via network adapter 660. Network adapter 660 can communicate with other modules of electronic device 600 via bus 630. It should be understood that, although not shown in the figures, other hardware and / or software modules can be used in conjunction with electronic device 600, including but not limited to: microcode, device drivers, redundant processing units, external disk drive arrays, RAID systems, tape drives, and data backup storage platforms.

[0150] This invention also provides a computer-readable storage medium for storing a program that, when executed, implements the steps of a neurovascular coupling imaging method. In some possible implementations, various aspects of the invention can also be implemented as a program product comprising program code that, when run on a terminal device, causes the terminal device to perform the steps described in the above-described method section of this specification according to various exemplary embodiments of the invention.

[0151] As shown above, the neurovascular coupling imaging system of this invention in this embodiment can integrate different visible light or near-infrared laser light sources into the optical path to be acquired, and is compatible with various imaging requirements such as visible light for nerve cell activity and near-infrared speckle imaging for non-invasive blood flow imaging. It ensures that there is no time difference between image acquisition and light source switching without losing time resolution, and facilitates subsequent data storage and calculation.

[0152] Figure 7 This is a schematic diagram of the structure of the computer-readable storage medium of the present invention. (Reference) Figure 7 As shown, a program product 800 for implementing the above-described method according to an embodiment of the present invention is described. This product may employ a portable compact disc read-only memory (CD-ROM) and include program code, and may run on a terminal device, such as a personal computer. However, the program product of the present invention is not limited thereto. In this document, the readable storage medium may be any tangible medium containing or storing a program that may be used by or in conjunction with an instruction execution system, apparatus, or device.

[0153] The program product may employ any combination of one or more readable media. A readable medium may be a readable signal medium or a readable storage medium. A readable storage medium may be, for example, but not limited to, an electrical, magnetic, optical, electromagnetic, near-infrared, or semiconductor system, apparatus, or device, or any combination thereof. More specific examples (a non-exhaustive list) of readable storage media include: electrical connections having one or more wires, portable disks, hard disks, random access memory (RAM), read-only memory (ROM), erasable programmable read-only memory (EPROM or flash memory), optical fiber, portable compact disk read-only memory (CD-ROM), optical storage devices, magnetic storage devices, or any suitable combination thereof.

[0154] Computer-readable storage media may include data signals propagated in baseband or as part of a carrier wave, carrying readable program code. Such propagated data signals may take various forms, including but not limited to electromagnetic signals, optical signals, or any suitable combination thereof. A readable storage medium may also be any readable medium other than a readable storage medium that can transmit, propagate, or transfer a program for use by or in connection with an instruction execution system, apparatus, or device. The program code contained on the readable storage medium may be transmitted using any suitable medium, including but not limited to wireless, wired, optical fiber, RF, etc., or any suitable combination thereof.

[0155] Program code for performing the operations of this invention can be written in any combination of one or more programming languages, including object-oriented programming languages ​​such as Java and C++, and conventional procedural programming languages ​​such as C or similar languages. The program code can execute entirely on the user's computing device, partially on the user's device, as a standalone software package, partially on the user's computing device and partially on a remote computing device, or entirely on a remote computing device or server. In cases involving remote computing devices, the remote computing device can be connected to the user's computing device via any type of network, including a local area network (LAN) or a wide area network (WAN), or it can be connected to an external computing device (e.g., via the Internet using an Internet service provider).

[0156] In summary, the purpose of this invention is to provide a neurovascular-based coupled imaging system, method, device, and storage medium that can integrate different visible light or near-infrared laser light sources into the optical path to be acquired, thus being compatible with various imaging requirements such as visible light for nerve cell activity and near-infrared speckle imaging for non-invasive blood flow imaging. This ensures that there is no time difference between image acquisition and light source switching without sacrificing time resolution, and facilitates subsequent data storage and calculation.

[0157] The above description, in conjunction with specific preferred embodiments, provides a further detailed explanation of the present invention. It should not be construed that the specific implementation of the present invention is limited to these descriptions. For those skilled in the art, various simple deductions or substitutions can be made without departing from the concept of the present invention, and all such modifications and substitutions should be considered within the scope of protection of the present invention.

Claims

1. A neurovascular-based coupled imaging device, characterized in that, include: An image acquisition device (5); A microscope frame (4) is positioned in the light-gathering direction of the image acquisition device (5); A visible light system emits visible light onto the sample after it is reflected by the first beam splitter in the microscope frame (4); A near-infrared light system emits near-infrared light onto the sample after it is reflected by the second beam splitter in the microscope frame (4); as well as A triggering device (3) switches the frequency of each light source in the visible light system and the near-infrared light system to emit light in a time-division manner based on the acquisition frequency of the image acquisition device (5); The acquisition frequency of the image acquisition device (5) includes a first high level and a second high level that are spaced apart from each other. The high level of the trigger emission in the working frequency of the visible light system overlaps with the first high level in time. The high level of the trigger emission in the working frequency of the near-infrared light system overlaps with the second high level in time. A visible light image set is established based on the first image acquired within the first high level, and a near-infrared light image set is established based on the second image acquired within the second high level. The first beam splitter (402) forms a 45° angle with the incident light formed by the beam emitted by the visible light source. The reflected light forms a 45° angle with the first beam splitter (402) and is perpendicular to the plane of the observation carrier carrying the sample. The second beam splitter (403) forms a 45° angle with the incident light formed by the beam emitted by the near-infrared laser source. The reflected light forms a 45° angle with the second beam splitter (403) and is perpendicular to the plane of the observation carrier carrying the sample. The plane of the first beam splitter (402) and the plane of the second beam splitter (403) are perpendicular to each other.

2. The neurovascular-based coupled imaging device as described in claim 1, characterized in that, It also includes a data processing module (6), which is connected to the image acquisition unit (5). Based on the high level of several different bands of visible light emitted in time division at the working frequency of the visible light system, it obtains a first high level set in time sequence. Based on the high level of at least one different band of near-infrared light emitted in time division at the working frequency of the near-infrared light system, it obtains a second high level set in time sequence. Based on the superposition of the first high level set and the second high level set in time sequence, it obtains a third high level set. The image acquisition unit (5) performs image acquisition based on the third high level set and classifies and stores the images in the data processing module (6) according to the mapping relationship between different bands and high level time sequence.

3. The neurovascular-based coupled imaging device as described in claim 1, characterized in that, The microscope frame (4) is disposed between the image acquisition device (5) and the observation carrier. The microscope frame (4) includes a filter (401), a first beam splitter (402), a second beam splitter (403) and an objective lens (404) arranged sequentially along the optical path. The visible light system is an LED light source system (1), which emits visible light of a preset wavelength into the first beam splitter (402), and after being reflected by the first beam splitter (402), it is emitted onto the object to be sampled; The near-infrared light system is a near-infrared laser source system (2), which emits near-infrared laser light into the second beam splitter (403), and after being reflected by the second beam splitter (403), it is emitted onto the object to be sampled; The triggering device (3) is connected to the image acquisition device (5), the LED light source system (1) and the near-infrared laser light source system (2) respectively. The triggering device (3) switches the first frequency of each light source in the LED light source system (1) and the near-infrared laser light source system (2) to be emitted in a time-division manner to match the second frequency of the triggering device (3) driving the image acquisition device (5) to acquire images.

4. The neurovascular-based coupled imaging device as described in claim 3, characterized in that, The LED light source system (1) is equipped with at least one narrowband visible light LED light source selected from the 405nm, 488nm, 535nm and 647nm bands; the first beam splitter (402) is a non-polarized wideband planar beam splitter, the beam splitting band of the first beam splitter (402) is 400 to 800nm, and the ratio of the projected frequency T to the reflectance R is 1:1; The second beam splitter (403) is a non-polarized wide-band planar beam splitter. The beam splitting band of the second beam splitter (403) is from 400 to 800 nm, and the ratio of the transmittance T to the reflectance R is 1:

1.

5. The neurovascular-based coupled imaging device as described in claim 3, characterized in that, The LED light source system (1) and the near-infrared laser light source system (2) are located on both sides of the microscope frame (4). The objective lens (404) is a combination of microscope slides with variable magnification and adjustable focal length. The observation carrier carries nerve and blood vessel tissue. The optical axis of the reflected light of the first beam splitter (402) and the optical axis of the reflected light of the second beam splitter (403) are coaxial with the optical axis of the incident light of the image acquisition device (5).

6. The neurovascular-based coupled imaging device as described in claim 3, characterized in that, The high level in the working timing pulse of the LED light source system (1) is the emission state, the high level in the working timing pulse of the near-infrared laser light source system (2) is the emission state, and the high level in the working timing pulse of the image acquisition device (5) is the acquisition state. The falling edge of the high level in the working timing pulse of the LED light source system (1) overlaps with the rising edge of the high level in the working timing pulse of the near-infrared laser light source system (2) in timing, and this overlapping timing overlaps with the low level between adjacent high levels in the working timing pulse of the image acquisition device (5) in timing.

7. A neurovascular-based coupling imaging method, employing the neurovascular-based coupling imaging device as described in any one of claims 1 to 6, characterized in that, include: S110. Based on the high level of at least one different band of visible light emitted in time-division multiplexing at the operating frequency of the visible light system, a first set of high levels in time sequence is established. S120. Based on the high level of at least one different band of near-infrared light emitted in time division at the operating frequency of the near-infrared light system, a second set of high levels is established in time sequence. S130. Obtain the third high-level set by superimposing the first high-level set and the second high-level set in timing. S140, The image acquisition device continuously acquires images based on the third high-level set; and S150. Obtain a visible light image sequence based on the first image acquired during the first high level, and obtain a near-infrared light image sequence based on the second image acquired during the second high level.

8. A neurovascular coupling imaging device, characterized in that, include: processor; A memory in which executable instructions of the processor are stored; The processor is configured to perform the steps of the neurovascular-based coupled imaging method of claim 7 by executing the executable instructions.

9. A computer-readable storage medium for storing a program, characterized in that, When the program is executed by the processor, it implements the steps of the neurovascular-based coupled imaging method of claim 7.

Images