Three-dimensional living body imaging system and method

Abstract

The invention relates to the technical field of biological imaging, in particular to a three-dimensional living body imaging system and method, and the system comprises an excitation device which is used for emitting a first parallel Gaussian light spot; the near-infrared structured light generation device is used for modulating the first parallel Gaussian light spot into first dynamic structured light; the first deformable mirror is used for reflecting a structural fluorescence signal in a living body to be imaged to the first dichroscope; the first dichroscope is used for dividing the structure fluorescence signal, reflected by the first deformable mirror, in the to-be-imaged living body into a first path of light and a second path of light; the near-infrared adaptive optical device is used for obtaining aberration according to the first path of light and a specified light structure; the imaging device is used for acquiring a two-dimensional image corresponding to the living body to be imaged based on the second path of light; and the three-dimensional reconstruction device is used for reconstructing the two-dimensional image into a three-dimensional model and controlling the first deformable mirror according to the aberration, so that the aberration between the first path of light and the specified light structure is reduced within a preset range.

Description

Technical Field

[0001] The present invention relates to the field of biological imaging technology, and in particular to a three-dimensional living body imaging system and method. Background Art

[0002] With the development of medical imaging technology, three-dimensional in vivo imaging systems have gained increasing attention in biomedical research and clinical applications. Traditional two-dimensional imaging technology is unable to provide information on spatial position, which limits its application in the study of internal structure and function of living bodies. Therefore, three-dimensional imaging technology has gradually become an important tool for in vivo disease research and clinical diagnosis. Especially in the field of biomedicine, three-dimensional in vivo imaging technology can provide more comprehensive and accurate tissue structure and function data, which is of great significance for disease diagnosis, treatment, efficacy evaluation and pathological research.

[0003] Photons in the second near-infrared region (1000nm–1700nm) have a high signal-to-noise ratio and resolution in biological tissue imaging because of their weak absorption, weak scattering, and weak autofluorescence characteristics in biological tissues. However, due to the limitations of existing wide-field imaging equipment and algorithms in the second near-infrared region, only two-dimensional imaging of living organisms can be achieved, and it is difficult to obtain three-dimensional wide-field structural and functional information, making it difficult to gain a deep understanding of the complex internal structure and functional state of living organisms.

[0004] At present, near-infrared in vivo imaging systems mainly rely on two-dimensional imaging technology and cannot directly achieve wide-field three-dimensional imaging. Traditional two-dimensional imaging technology can only provide partial information inside the living body, but cannot fully display the three-dimensional information of the living body structure. This poses a great challenge to deep tissue research and functional diagnosis inside the living body. Summary of the invention

[0005] In view of the above-mentioned shortcomings and deficiencies of the prior art, the present invention provides a three-dimensional in vivo imaging system and method, which solves the technical problem that wide-field three-dimensional in vivo imaging is difficult to achieve in the prior art.

[0006] In order to achieve the above object, the main technical solutions adopted by the present invention include:

[0007] In a first aspect, an embodiment of the present invention provides a three-dimensional in vivo imaging system, the system comprising:

[0008] An exciting device, used for emitting a first parallel Gaussian light spot;

[0009] A near-infrared structured light generating device, used for modulating the first parallel Gaussian light spot into a first dynamic structured light of a specified light structure;

[0010] A first deformable mirror, used for reflecting the structural fluorescence signal inside the living body to be imaged to the first dichroic mirror;

[0011] The structured fluorescence signal inside the living body to be imaged includes the fluorescence generated inside the living body to be imaged and the first dynamic structured light reflected by the surface of the living body to be imaged after the first dynamic structured light is irradiated into the living body to be imaged;

[0012] The fluorescence generated inside the living body to be imaged includes: a first dynamic structure fluorescence and a second dynamic structure fluorescence generated when the first dynamic structure light is irradiated inside the living body to be imaged;

[0013] A first dichroic mirror, used for separating the structural fluorescence signal inside the living body to be imaged reflected by the first deformable mirror into a first light path entering the near-infrared adaptive optical device and a second light path entering the imaging device;

[0014] The first light path only includes the first dynamic structure fluorescence emitted from the living body to be imaged; the second light path only includes the second dynamic structure fluorescence generated from the living body to be imaged;

[0015] A near-infrared adaptive optical device, used for obtaining an aberration between the first light path and the specified light structure according to the first light path and the specified light structure;

[0016] An imaging device, used for acquiring a two-dimensional image corresponding to the living body to be imaged based on the second light;

[0017] A three-dimensional reconstruction device is used to reconstruct the two-dimensional image corresponding to the living body to be imaged into a three-dimensional model, and is used to control the first deformable mirror according to the aberration between the first light and the specified light structure so that the aberration between the first light and the specified light structure is reduced to within a preset range.

[0018] Preferably, the excitation device comprises:

[0019] A first near-infrared laser device, configured to emit a first light beam;

[0020] a second near-infrared laser device, configured to emit a second light beam;

[0021] A second dichroic mirror, used for combining the first light beam and the second light beam into a first parallel Gaussian light spot;

[0022] The first near-infrared laser device comprises:

[0023] A first near-infrared laser for emitting a first laser and a first convex lens assembly for collimating the first laser and expanding the first laser into a first light beam with a diameter greater than 15 mm; the first convex lens assembly includes a first convex lens and a second convex lens;

[0024] The wavelength of the first laser is 808 nanometers and the power is 5 watts;

[0025] The first dynamic structure fluorescence generated inside the living body to be imaged is generated inside the living body to be imaged after the first light beam in the first parallel Gaussian light spot passes through the near-infrared structure light generating device and then irradiates the living body to be imaged;

[0026] The second near-infrared laser device comprises:

[0027] A second near-infrared laser for emitting a second laser and a second convex lens assembly for collimating the second laser and expanding the second laser into a second light beam with a diameter greater than 15 mm; the second convex lens assembly includes a third convex lens and a fourth convex lens;

[0028] The wavelength of the second laser is 980 nanometers and the power is 10 watts;

[0029] The second dynamic structure fluorescence generated inside the living body to be imaged is generated inside the living body to be imaged after the second light beam in the first parallel Gaussian spot passes through the near-infrared structure light generating device and then irradiates the living body to be imaged.

[0030] Preferably, the near-infrared structured light generating device is a near-infrared spatial light modulator;

[0031] Among them, the pixel array size of the near-infrared spatial light modulator is 912×1140, the maximum structured light refresh frequency is 120Hz, and the applicable wavelength range of the near-infrared spatial light modulator is 700nm to 2500nm.

[0032] Preferably, the first deformable mirror is mounted on a five-axis translation stage;

[0033] The first deformable mirror has 69 silver mirrors with actuators;

[0034] Each silver mirror can adjust its reflection angle, and its reflection angle is less than or equal to 15 degrees.

[0035] Preferably, the near-infrared adaptive optical device comprises a third convex lens component for converging the first light path, a first long-pass filter, and a wavefront sensor for obtaining the aberration between the first light path and the designated light structure based on the first light path passing through the third convex lens component, the first long-pass filter, and the designated light structure in sequence;

[0036] The wavelength range of the first long-pass filter is 1000nm to 1500nm; wherein the wavefront sensor is arranged on a three-axis translation stage.

[0037] Preferably, the imaging device comprises: a second long-pass filter, an imaging lens, and a first near-infrared camera arranged in sequence;

[0038] The wavelength range of the second long pass filter is 1000nm to 1500nm;

[0039] The first near-infrared camera is an InGaAs near-infrared camera with a resolution of 640×512 pixels.

[0040] Preferably, all convex lenses have an optical size of 1 inch in diameter.

[0041] Preferably, the living body to be imaged is pre-processed, wherein the pre-processing method of the living body to be imaged is:

[0042] The first fluorescent probe is injected into the living body to be imaged through the tail vein or injected into the living body to be imaged through gavage;

[0043] The first fluorescent probe is a fluorescent probe that can be excited by near-infrared lasers of 808 nanometers and 980 nanometers and has an emission wavelength between 1000 nanometers and 1700 nanometers.

[0044] Preferably, the three-dimensional reconstruction device uses MatLab image processing software to reconstruct the two-dimensional image corresponding to the living body to be imaged into a three-dimensional model;

[0045] The three-dimensional reconstruction device uses Alpao Core Engine software to control the first deformable mirror according to the aberration between the first light and the specified light structure, so as to reduce the aberration between the first light and the specified light structure within a preset range.

[0046] On the other hand, this embodiment further provides a three-dimensional in vivo imaging method, which is performed by the three-dimensional in vivo imaging system described in the first aspect, and includes:

[0047] S1, placing a standard chessboard positioning plate in the field of view of a first near-infrared camera, using a near-infrared structured light generating device to generate a first dynamic structured light and obtaining a two-dimensional image of the standard chessboard positioning plate in the first near-infrared camera, and then calibrating a reference plane for three-dimensional imaging of a living body to be imaged in a three-dimensional reconstruction device;

[0048] S2, placing the living body to be imaged on a calibrated reference plane, using a first parallel Gaussian light spot emitted by an excitation device, and then a near-infrared structured light generating device to modulate the first parallel Gaussian light spot into a first dynamic structured light with the same specified light structure, and irradiating the first dynamic structured light onto the living body to be imaged;

[0049] S3, a first deformable mirror, used to reflect the structural fluorescence signal inside the living body to be imaged to the first dichroic mirror; the structural fluorescence signal inside the living body to be imaged includes the fluorescence generated inside the living body to be imaged and the first dynamic structured light reflected by the surface of the living body to be imaged after the first dynamic structured light is irradiated into the living body to be imaged;

[0050] The fluorescence generated inside the living body to be imaged includes: a first dynamic structure fluorescence and a second dynamic structure fluorescence generated when the first dynamic structure light is irradiated inside the living body to be imaged;

[0051] A first dichroic mirror, used for separating the structural fluorescence signal inside the living body to be imaged reflected by the first deformable mirror into a first light path entering the near-infrared adaptive optical device and a second light path entering the imaging device;

[0052] The first light path only includes the first dynamic structure fluorescence emitted from the living body to be imaged; the second light path only includes the second dynamic structure fluorescence generated from the living body to be imaged;

[0053] A near-infrared adaptive optical device, used for obtaining an aberration between the first light path and the specified light structure according to the first light path and the specified light structure;

[0054] An imaging device, used for acquiring a two-dimensional image corresponding to the living body to be imaged based on the second light;

[0055] A three-dimensional reconstruction device is used to reconstruct the two-dimensional image corresponding to the living body to be imaged into a three-dimensional model, and is used to control the first deformable mirror according to the aberration between the first light and the specified light structure so that the aberration between the first light and the specified light structure is reduced to within a preset range.

[0056] The beneficial effects of the present invention are:

[0057] A three-dimensional in vivo imaging system of the present invention adopts the following technical measures:

[0058] Excitation device: emits a first parallel Gaussian light spot to enhance the interaction effect between the light and the living body to be imaged. In this way, the intensity and uniformity of the light excitation signal can be improved, which helps to better excite the fluorescence inside the living body to be imaged and provide a clearer excitation signal.

[0059] Near-infrared structured light generation device: modulates the first parallel Gaussian light spot into a dynamic structured light with a specified light structure, which can provide higher information density and spatial resolution. Near-infrared light (wavelength 1000nm-1700nm) has weak scattering, weak absorption and weak autofluorescence characteristics, and has a good signal-to-noise ratio and resolution in in vivo imaging, so the use of near-infrared light helps to enhance imaging quality.

[0060] The first deformable mirror is used to reflect the structural fluorescence signal inside the living body to be imaged onto the first dichroic mirror. This mirror can effectively process the first dynamic structure fluorescence and the second dynamic structure fluorescence separately to avoid aliasing and ensure the purity and accuracy of the imaging signal. This ensures that the light signals of each component are not mixed, thereby improving the accuracy of imaging.

[0061] First dichroic mirror: divides the reflected structural fluorescence signal into the first light path entering the near-infrared adaptive optical device and the second light path entering the imaging device. Through this separation technology, the imaging processing of the first dynamic structure fluorescence and the second dynamic structure fluorescence can be optimized separately, which helps to improve the contrast and detail display effect of the image. The first light path mainly includes the first dynamic structure fluorescence emitted from the inside of the living body to be imaged. The imaging of this part of light can provide delicate structural information inside the living body, while the second light path mainly includes the second dynamic structure fluorescence, which helps to capture the light signal reflected from the surface and further enrich the imaging content.

[0062] Near-infrared adaptive optical device: adjust the position of the first deformable mirror according to the first light path and the specified light structure to reduce aberrations and improve imaging precision and accuracy. By adjusting the position of the deformable mirror, the aberrations in the light path can be compensated in real time to ensure that the light spot of each light path is uniform and clear, thereby improving the quality and resolution of the image. Adaptive optical technology can improve the quality and stability of imaging by correcting the distortion and deviation in the optical system through real-time feedback, making the imaging effect closer to the real living structure.

[0063] Imaging device: A two-dimensional image is obtained based on the second light. Such processing helps to generate detailed spatial information corresponding to the living body to be imaged. The two-dimensional image captures the optical signals of the surface and internal structure, which can provide basic data for subsequent three-dimensional reconstruction, while improving the image's detail display and contrast.

[0064] 3D reconstruction device: Use 2D images to reconstruct the 3D model of the living body to be imaged, providing more comprehensive and accurate spatial information. 3D reconstruction can show the internal structure and spatial distribution of the living body, and provide deeper anatomical information and functional data. The position of the first deformable mirror is adjusted to optimize the imaging and ensure the accuracy and stability of the reconstructed model. This process makes it possible to make the leap from 2D to 3D in vivo imaging by dynamically adjusting the optical path and the position of the deformable mirror, greatly improving the spatial resolution and accuracy of imaging.

[0065] Compared with the prior art, the system of the present invention makes it possible to make the leap from two-dimensional to three-dimensional in vivo imaging by combining dynamic structured light and adaptive optics technology, greatly improving the spatial resolution and accuracy of imaging. The system can obtain the internal structure information of the living body to be imaged in real time and accurately, meeting the needs of biomedical research and clinical applications for high-resolution, three-dimensional imaging. At the same time, the system ensures the stability and high-quality output of imaging by dynamically adjusting the optical path and deformable mirror. BRIEF DESCRIPTION OF THE DRAWINGS

[0066] Figure 1 It is a schematic diagram of the structure of a three-dimensional in vivo imaging system of the present invention;

[0067] Figure 2 The figure is a flow chart of a three-dimensional in vivo imaging method of the present invention.

[0068] Description of Reference Numerals

[0069] 1: excitation device; 11: first near-infrared laser device; 111: first near-infrared laser; 112: first convex lens assembly; 12: second near-infrared laser device; 121: second near-infrared laser; 122: second convex lens assembly; 13: second dichroic mirror; 2: near-infrared structured light generating device; 3: first deformable mirror; 4: first dichroic mirror; 5: near-infrared adaptive optical device; 51: third convex lens assembly; 52: first long-pass filter; 53: wavefront sensor; 6: imaging device; 61: second long-pass filter; 62: imaging lens; 63: first near-infrared camera; 7: three-dimensional reconstruction device. DETAILED DESCRIPTION

[0070] In order to better explain the present invention and facilitate understanding, the present invention is described in detail below through specific implementation modes in conjunction with the accompanying drawings.

[0071] In order to better understand the above technical solution, exemplary embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although exemplary embodiments of the present invention are shown in the accompanying drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided to enable a clearer and more thorough understanding of the present invention and to fully convey the scope of the present invention to those skilled in the art.

[0072] Embodiment 1

[0073] See also Figure 1 This embodiment provides a three-dimensional in vivo imaging system, the system comprising:

[0074] Excitation device 1, used for emitting a first parallel Gaussian light spot;

[0075] In this embodiment, the excitation device 1 includes:

[0076] A first near-infrared laser device 11, used for emitting a first light beam;

[0077] A second near-infrared laser device 12, used to emit a second light beam;

[0078] A second dichroic mirror 13, used for combining the first light beam and the second light beam into a first parallel Gaussian light spot;

[0079] The first near-infrared laser device 11 comprises:

[0080] A first near-infrared laser 111 for emitting a first laser and a first convex lens assembly 112 for collimating the first laser and expanding the first laser into a first light beam with a diameter greater than 15 mm; the first convex lens assembly includes a first convex lens and a second convex lens; the diameter of the first light beam after expansion is greater than 15 mm, which can evenly cover the imaging area, reduce the unevenness of the laser spot, and improve the uniformity and irradiation area of ​​the excitation light. In this way, more uniform laser irradiation can be achieved in the imaging area, avoiding local excessive irradiation and light damage, and improving the stability and quality of imaging.

[0081] The wavelength of the first laser is 808 nanometers and the power is 5 watts; this wavelength (808 nanometers) belongs to the near-infrared spectral region (800-900nm), has low light scattering characteristics and less fluorescence background interference, and is an ideal light source choice for in vivo imaging. Its high power (5 watts) can effectively excite fluorescent molecules inside the living body, such as oxygenated hemoglobin or calcium ion fluorescein, enhance the intensity of the fluorescence signal, and provide high-quality excitation signals. This helps to improve the signal-to-noise ratio of imaging, thereby enhancing the contrast and resolution of imaging.

[0082] The first dynamic structure fluorescence generated inside the living body to be imaged is generated inside the living body to be imaged after the first light beam in the first parallel Gaussian light spot passes through the near-infrared structure light generating device 2 and then irradiates the living body to be imaged;

[0083] In this embodiment, by merging the first light beam and the second light beam, the advantages of dual-wavelength laser can be utilized to improve the diversity and flexibility of the imaging system. Dual-wavelength light can selectively excite and detect different fluorescent molecules, thereby improving the efficiency and flexibility of multi-channel fluorescence imaging. In addition, the first parallel Gaussian light spot formed after merging can evenly illuminate the imaging area, enhance the energy distribution of the laser, and further improve the signal-to-noise ratio and imaging quality of the imaging.

[0084] The second near-infrared laser device 12 comprises:

[0085] A second near-infrared laser 121 for emitting a second laser and a second convex lens assembly 122 for collimating the second laser and expanding the second laser into a second light beam with a diameter greater than 15 mm; the second convex lens assembly includes a third convex lens and a fourth convex lens;

[0086] The second laser has a wavelength of 980 nanometers and a power of 10 watts. Near-infrared light with a wavelength of 980 nanometers has a deeper penetration ability than light with a wavelength of 808 nanometers and can penetrate deeper into living tissues, which is very beneficial for imaging deeper tissues. High-power (10-watt) lasers can provide powerful excitation energy, which helps to excite fluorescent markers such as fat-soluble pigments or other near-infrared probes and enhance imaging signals. By utilizing dual-wavelength excitation of 980 nanometer lasers and 808 nanometer lasers, the imaging depth can be increased and the excitation and detection efficiency of different fluorescent molecules can be improved.

[0087] The second dynamic structure fluorescence generated inside the living body to be imaged is generated inside the living body to be imaged after the second light beam in the first parallel Gaussian spot passes through the near-infrared structure light generating device 2 and then irradiates the living body to be imaged.

[0088] The near-infrared structured light generating device 2 is used to modulate the first parallel Gaussian light spot into a first dynamic structured light of a specified light structure; in this embodiment, the modulated dynamic structured light can improve the imaging accuracy and clarity of the internal structure of the tissue, help identify complex tissue structures and functional changes, and enhance the depth and resolution of in vivo imaging.

[0089] The first deformable mirror 3 is used to reflect the structural fluorescence signal inside the living body to be imaged to the first dichroic mirror 4; the coordinated use of the first deformable mirror 3 and the first dichroic mirror 4 helps to separate the optical path and optically adjust the structural fluorescence signal. The first deformable mirror 3 reflects the fluorescence signal, and the first dichroic mirror 4 separates it into the optical path entering the near-infrared adaptive optical device 5 and the imaging device 6. This structure helps to separate the fluorescence signal, so that the near-infrared adaptive optical device can optimize the optical path aberration and improve the clarity and accuracy of imaging. At the same time, the separated optical path avoids mutual interference between the fluorescence signals, and improves the imaging quality and imaging depth.

[0090] The structured fluorescence signal inside the living body to be imaged includes the fluorescence generated inside the living body to be imaged and the first dynamic structured light reflected by the surface of the living body to be imaged after the first dynamic structured light is irradiated into the living body to be imaged;

[0091] The fluorescence generated inside the living body to be imaged includes: a first dynamic structure fluorescence and a second dynamic structure fluorescence generated when the first dynamic structure light is irradiated inside the living body to be imaged;

[0092] A first dichroic mirror 4 is used to separate the structural fluorescence signal inside the living body to be imaged reflected by the first deformable mirror 3 into a first light path entering the near-infrared adaptive optical device 5 and a second light path entering the imaging device 6;

[0093] The first light path only includes the first dynamic structure fluorescence emitted from the living body to be imaged; the second light path only includes the second dynamic structure fluorescence generated from the living body to be imaged;

[0094] The near-infrared adaptive optical device 5 is used to obtain the aberration between the first light path and the specified light structure according to the first light path and the specified light structure; the near-infrared adaptive optical device 5 can enhance the imaging accuracy of the optical system by adjusting the aberration.

[0095] The imaging device 6 is used to obtain a two-dimensional image corresponding to the living body to be imaged based on the second light. The imaging device 6 obtains two-dimensional images corresponding to the living body to be imaged, and these images can be used as a basis for further analysis and three-dimensional reconstruction.

[0096] The three-dimensional reconstruction device 7 is used to reconstruct the two-dimensional image corresponding to the living body to be imaged into a three-dimensional model, and to control the first deformable mirror 3 according to the aberration between the first light and the specified light structure so that the aberration between the first light and the specified light structure is reduced to within a preset range.

[0097] The near-infrared structured light generating device 2 is a near-infrared spatial light modulator;

[0098] Among them, the pixel array size of the near-infrared spatial light modulator is 912×1140, the maximum structured light refresh frequency is 120Hz, and the applicable wavelength range of the near-infrared spatial light modulator is 700nm to 2500nm.

[0099] In this embodiment, the high pixel array size (912×1140) of the near-infrared spatial light modulator can provide finer spatial modulation capabilities, so that the generated structured light has a higher resolution. This feature helps to more accurately control the spatial distribution and modulation of light in in vivo imaging, thereby enhancing the fineness and contrast of imaging. The maximum refresh rate of the near-infrared spatial light modulator is 120Hz, which enables the system to dynamically adjust the structured light in real time and track and record rapid changes inside the living body. In in vivo imaging, real-time dynamic response capabilities are crucial to better capture the dynamic changes in tissue structure and functional state. The near-infrared spatial light modulator is suitable for a wavelength range of 700nm to 2500nm, which enables the system to adapt to the excitation and imaging needs of a variety of fluorescent markers. This wide-band adaptability enhances the flexibility of the system and can be used in different imaging applications to meet diverse imaging requirements.

[0100] In the practical application of this embodiment, the first deformable mirror 3 is mounted on a five-axis translation platform;

[0101] The first deformable mirror 3 has 69 silver mirrors with actuators;

[0102] Each silver mirror can adjust its reflection angle, and its reflection angle is less than or equal to 15 degrees.

[0103] The first deformable mirror 3 is mounted on a five-axis translation stage, which can achieve precise fine-tuning and dynamic adjustment in multiple directions. The five-axis degrees of freedom enable the deformable mirror to be adjusted in three spatial directions (X, Y, Z) and two angles (pitch and roll), so as to adapt to the rapid changes and complex structures inside the living body, effectively compensate for optical distortion and achieve real-time adjustment.

[0104] In this embodiment, each silver mirror can adjust its reflection angle, and the reflection angle is less than or equal to 15 degrees. Such a design can achieve more precise light beam control. Adjusting the angle can effectively change the reflection path of light, thereby achieving more precise control of the imaging light beam.

[0105] Specifically, the near-infrared adaptive optical device 5 includes a third convex lens component 51 for converging the first light path, a first long-pass filter 52, and a wavefront sensor 53 for obtaining the aberration between the first light path and the specified light structure based on the first light path and the specified light structure passing through the third convex lens component 51 and the first long-pass filter 52 in sequence;

[0106] The wavelength range of the first long-pass filter 52 is 1000 nm to 1500 nm; wherein the wavefront sensor 53 is arranged on a three-axis translation stage.

[0107] The near-infrared adaptive optical device 5 includes a third convex lens component 51 and a first long-pass filter 52 for reducing the first path of light. Through the configuration of these components, the path and transmission characteristics of the light beam can be accurately controlled to ensure the quality and stability of the light beam. The third convex lens component 51 helps to collimate and expand the light beam to the required diameter, providing a uniform light irradiation area; the first long-pass filter 52 selectively transmits light of a specific wavelength, improves spectral selectivity, effectively filters out unnecessary light wave interference, and improves the optical efficiency of the system. The wavefront sensor 53 is set on a three-axis translation stage, and can monitor the aberration between the first path of light and the specified light structure in real time. The degree of freedom of the three-axis translation stage enables the wavefront sensor to detect and adjust the aberration of the optical system in multiple directions, and provide real-time feedback and optimization of optical performance. The wavefront sensor 53 can accurately detect the changes and aberrations of the light wave front, and compensate for these changes by adjusting the position of the optical element in real time, ensuring stable transmission of the light beam and enhancing the imaging quality. This helps to improve the imaging quality and resolution of the imaging system, especially in dynamic and complex imaging environments. By setting the wavefront sensor on the three-axis translation stage, the aberration in the light beam transmission path can be monitored in real time and dynamically adjusted. This design enables the near-infrared adaptive optical device to adapt to the complex changes in the optical environment inside the living body in real time, and meet the needs of real-time three-dimensional imaging and detection of the living body.

[0108] The imaging device 6 includes: a second long-pass filter 61, an imaging lens 62, and a first near-infrared camera 63 arranged in sequence;

[0109] The wavelength range of the second long pass filter 61 is 1000nm to 1500nm;

[0110] The first near-infrared camera 63 is an InGaAs near-infrared camera with a resolution of 640×512 pixels.

[0111] The second long-pass filter 61 in the imaging device 6 helps to selectively transmit a spectrum of a specific wavelength. The wavelength range is 1000nm to 1500nm, which effectively reduces background noise and interference light, ensuring that only useful near-infrared fluorescence signals are transmitted. This improves the spectral selectivity of the system and enhances the imaging quality. The second long-pass filter 61 can effectively filter out spectral interference of other wavelengths, provide a clear fluorescence image, and help enhance the signal-to-noise ratio and imaging quality.

[0112] The first near-infrared camera 63 uses an InGaAs detector with high resolution (640×512 pixels) to capture fine optical details. The combination with the InGaAs detector, especially in the near-infrared spectrum, its spectral sensitivity and response speed can provide high-quality near-infrared imaging data. This high-resolution imaging device can clearly capture the details of the internal structure of the living body and improve the imaging quality and contrast, which is very important for dynamic living body imaging and deep tissue imaging, and helps to accurately diagnose and study physiological and pathological changes in the living body.

[0113] In this embodiment, the optical size of all convex lenses is 1 inch in diameter.

[0114] In this embodiment, the optical size of the convex lenses is uniform, which helps to ensure the consistency of the optical paths between the various optical elements of the system. The diameters of all convex lenses are consistent, ensuring that the light beam does not refract, deflect or diffract during transmission, thereby providing stable optical performance and imaging quality. The uniform optical size can improve the optical efficiency and imaging stability of the system, reduce the risk of spot deformation and beam distortion, and help achieve high-quality imaging effects.

[0115] It should be noted that the living body to be imaged is pre-processed, wherein the pre-processing method of the living body to be imaged is:

[0116] The first fluorescent probe is injected into the living body to be imaged through the tail vein or injected into the living body to be imaged through gavage;

[0117] The first fluorescent probe is a fluorescent probe that can be excited by 808 nanometers and 980 nanometers near-infrared lasers to produce first dynamic structure fluorescence and second dynamic structure fluorescence respectively, and has an emission wavelength between 1000 nanometers and 1700 nanometers.

[0118] By injecting the first fluorescent probe into the living body to be imaged, the intensity of the fluorescent signal can be enhanced. These probes can be excited by near-infrared lasers of 808 nanometers and 980 nanometers. These wavelength light sources can penetrate deeply into living tissues, improve the effect of exciting fluorescence, and reduce light scattering, thereby improving the contrast and signal-to-noise ratio of imaging. The enhanced signal intensity helps to obtain clearer two-dimensional and three-dimensional imaging data and improve the resolution and accuracy of imaging. Near-infrared light wavelengths of 808 nanometers and 980 nanometers have lower light scattering characteristics and can penetrate deeply into living tissues to deeper levels. By using light excitation probes of these wavelengths, the internal structures of the living body to be imaged, especially deep tissues, can be imaged at a deeper level, avoiding interference from surface light spots and improving the depth and targeting of imaging.

[0119] Specifically, the three-dimensional reconstruction device 7 uses MatLab image processing software to reconstruct the two-dimensional image corresponding to the living body to be imaged into a three-dimensional model;

[0120] In the present embodiment, efficient two-dimensional to three-dimensional image reconstruction can be achieved using MatLab's image processing software. This method utilizes MatLab's powerful processing power and image processing algorithm to quickly generate a clear and accurate three-dimensional model. By processing two-dimensional image data, depth information can be extracted and converted into a three-dimensional model, providing more intuitive and in-depth biological structure information. Rapid and accurate three-dimensional reconstruction is achieved, which helps to deeply understand and analyze the morphological characteristics and functional status of the internal structure of the living body. Especially in the field of medical imaging, it can be used for diagnosis, monitoring disease progression and evaluating treatment effects.

[0121] The three-dimensional reconstruction device 7 uses Alpao Core Engine software to control the first deformable mirror 3 according to the aberration between the first light and the specified light structure, so as to reduce the aberration between the first light and the specified light structure within a preset range.

[0122] In this embodiment, the Alpao Core Engine software automatically adjusts the reflection angle of the first deformable mirror by analyzing the aberration between the first light path and the specified light structure. This can accurately control the propagation direction and focus of the light, thereby reducing aberrations and improving the clarity of the imaging. By adjusting the light path through real-time electronic control, the beam quality and imaging effect can be optimized to ensure that the imaging system can still maintain stability and high-quality imaging performance in a dynamic environment, thereby presenting three-dimensional in vivo imaging results in real time and dynamically. This automated control mechanism can continuously optimize optical performance under different imaging conditions, help improve the adaptability and reliability of the imaging system, reduce human operation errors, and ensure high-quality three-dimensional imaging data.

[0123] Embodiment 2

[0124] See also Figure 2 This embodiment provides a three-dimensional in vivo imaging method, which is performed by the three-dimensional in vivo imaging system described in Embodiment 1, and includes:

[0125] S1, placing a standard chessboard positioning plate in the field of view of a first near-infrared camera 63, using a near-infrared structured light generating device 2 to generate a first dynamic structured light and obtaining a two-dimensional image of the standard chessboard positioning plate in the first near-infrared camera 63, and then calibrating a reference plane for three-dimensional imaging of a living body to be imaged in a three-dimensional reconstruction device 7;

[0126] In this embodiment, a standard chessboard positioning plate is placed in the field of view of the first near-infrared camera, and a near-infrared structured light generating device is used to generate dynamic structured light. This method can obtain a two-dimensional image of the chessboard positioning plate, thereby accurately calibrating the reference plane. This ensures that the imaging system can accurately map the imaging data to a standard three-dimensional space coordinate system in subsequent steps, which helps to improve the accuracy and consistency of imaging. It provides a stable reference benchmark for three-dimensional imaging of the living body to be imaged, reduces image distortion or offset caused by environmental changes or equipment errors during the imaging process, and improves the final imaging quality.

[0127] S2, placing the living body to be imaged on a calibrated reference plane, using the first parallel Gaussian light spot emitted by the excitation device 1, and then the near-infrared structured light generating device 2, modulating the first parallel Gaussian light spot into a first dynamic structured light with the same light structure as the specified light, and irradiating the first dynamic structured light onto the living body to be imaged;

[0128] In this embodiment, a parallel Gaussian light spot emitted by an excitation device is used, and is modulated into a specified dynamic structured light by a near-infrared structured light generating device, and irradiated onto the living body to be imaged. This dynamic structured light combined with laser excitation technology can effectively excite the fluorescence signal inside the living body to be imaged, including the first dynamic structure fluorescence and the second dynamic structure fluorescence. This method can accurately control the excitation and response of the fluorescence signal, so that the imaging system can observe clearer structural information inside the living body to be imaged. Through the fine control of the dynamic structured light, the contrast and signal-to-noise ratio of the fluorescence signal are enhanced, ensuring the clarity and accuracy of the imaging results, which has important application value for studying the internal structure of the living body.

[0129] S3, a first deformable mirror 3, reflecting the structural fluorescence signal inside the living body to be imaged to the first dichroic mirror 4; the structural fluorescence signal inside the living body to be imaged includes the fluorescence generated inside the living body to be imaged and the first dynamic structured light reflected by the surface of the living body to be imaged after the first dynamic structured light is irradiated into the living body to be imaged;

[0130] The fluorescence generated inside the living body to be imaged includes: a first dynamic structure fluorescence and a second dynamic structure fluorescence generated when the first dynamic structure light is irradiated inside the living body to be imaged;

[0131] In this embodiment, mutual interference of fluorescent signals can be effectively avoided by optical path separation, which helps to obtain purer imaging signals, thereby improving the quality of imaging results.

[0132] A first dichroic mirror 4 is used to separate the structural fluorescence signal inside the living body to be imaged reflected by the first deformable mirror 3 into a first light path entering the near-infrared adaptive optical device 5 and a second light path entering the imaging device 6;

[0133] The first light path only includes the first dynamic structure fluorescence emitted from the living body to be imaged; the second light path only includes the second dynamic structure fluorescence generated from the living body to be imaged;

[0134] A near-infrared adaptive optical device 5, used for obtaining the aberration between the first light path and the specified light structure according to the first light path and the specified light structure;

[0135] An imaging device 6, based on the second light path, acquires a two-dimensional image corresponding to the living body to be imaged;

[0136] The three-dimensional reconstruction device 7 reconstructs the two-dimensional image corresponding to the living body to be imaged into a three-dimensional model, and is used to control the first deformable mirror 3 according to the aberration between the first light and the specified light structure so that the aberration between the first light and the specified light structure is reduced within a preset range.

[0137] This embodiment provides a three-dimensional in vivo imaging method that uses a standard chessboard positioning plate to calibrate the reference plane, which can reduce the deviation caused by equipment errors and environmental changes during the imaging process, thereby improving the imaging accuracy and clarity. By using a first dichroic mirror to divide the reflected structural fluorescence signal into two light paths, the mutual interference between the fluorescence signals can be reduced, ensuring the purity and accuracy of the imaging results. The first dynamic structure fluorescence focused on by the first light path reduces the aberration of the first dynamic structure fluorescence and the specified light structure based on the adaptive system, and the second light path focuses on the aberration-free imaging of the second dynamic structure fluorescence, which is helpful for analyzing biological information at different depths inside the living body. The near-infrared adaptive optical device can automatically adjust the reflection angle of the first deformable mirror, compensate for aberrations in real time, optimize the optical imaging quality, and ensure that high-quality real-time dynamic imaging results can be obtained under various imaging conditions.

[0138] The three-dimensional reconstruction device reconstructs two-dimensional images into three-dimensional models, allowing researchers to intuitively observe and analyze the three-dimensional structure of the living organism to be imaged.

[0139] In the description of the present invention, it should be understood that the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of the features. In the description of the present invention, the meaning of "plurality" is two or more, unless otherwise clearly and specifically defined.

[0140] In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium; it can be the internal connection of two elements or the interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0141] In the present invention, unless otherwise clearly specified and limited, when a first feature is “on” or “below” a second feature, it may be that the first and second features are in direct contact, or that the first and second features are in indirect contact through an intermediate medium. Moreover, when a first feature is “above”, “above” or “above” a second feature, it may be that the first feature is directly above or obliquely above the second feature, or it may simply mean that the first feature is higher in level than the second feature. When a first feature is “below”, “below” or “below” a second feature, it may be that the first feature is directly below or obliquely below the second feature, or it may simply mean that the first feature is lower in level than the second feature.

[0142] In the description of this specification, the description of the terms "one embodiment", "some embodiments", "embodiment", "example", "specific example" or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, unless they are contradictory.

[0143] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may alter, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A three-dimensional in vivo imaging system, characterized in that: The system comprises: An exciting device (1) for emitting a first parallel Gaussian light spot; A near-infrared structured light generating device (2), used for modulating the first parallel Gaussian light spot into a first dynamic structured light of a specified light structure; A first deformable mirror (3) is used to reflect the structural fluorescence signal inside the living body to be imaged to a first dichroic mirror (4); The structured fluorescence signal inside the living body to be imaged includes the fluorescence generated inside the living body to be imaged and the first dynamic structured light reflected by the surface of the living body to be imaged after the first dynamic structured light is irradiated into the living body to be imaged; The fluorescence generated inside the living body to be imaged includes: a first dynamic structure fluorescence and a second dynamic structure fluorescence generated when the first dynamic structure light is irradiated inside the living body to be imaged; A first dichroic mirror (4) is used to separate the structural fluorescence signal inside the living body to be imaged, which is reflected by the first deformable mirror (3), into a first light path that enters a near-infrared adaptive optical device (5) and a second light path that enters an imaging device (6); The first light path only includes the first dynamic structure fluorescence emitted from the living body to be imaged; the second light path only includes the second dynamic structure fluorescence generated from the living body to be imaged; A near-infrared adaptive optical device (5), used for obtaining the aberration between the first light path and the specified light structure according to the first light path and the specified light structure; An imaging device (6) is used to acquire a two-dimensional image corresponding to the living body to be imaged based on the second light path; A three-dimensional reconstruction device (7) is used to reconstruct a two-dimensional image corresponding to the living body to be imaged into a three-dimensional model, and is used to control the first deformable mirror (3) according to the aberration between the first light path and the specified light structure, so that the aberration between the first light path and the specified light structure is reduced to within a preset range.

2. The three-dimensional in vivo imaging system according to claim 1, characterized in that: The excitation device (1) comprises: A first near-infrared laser device (11), configured to emit a first light beam; A second near-infrared laser device (12), configured to emit a second light beam; A second dichroic mirror (13), used for combining the first light beam and the second light beam into a first parallel Gaussian light spot; The first near-infrared laser device (11) comprises: A first near-infrared laser (111) for emitting a first laser and a first convex lens component (112) for collimating the first laser and expanding the first laser into a first light beam with a diameter greater than 15 mm; the first convex lens component comprises a first convex lens and a second convex lens; The wavelength of the first laser is 808 nanometers and the power is 5 watts; The first dynamic structure fluorescence generated inside the living body to be imaged is generated inside the living body to be imaged after the first light beam in the first parallel Gaussian light spot passes through the near-infrared structure light generating device (2) and then irradiates the living body to be imaged; The second near-infrared laser device (12) comprises: A second near-infrared laser (121) for emitting a second laser and a second convex lens assembly (122) for collimating the second laser and expanding the second laser into a second light beam with a diameter greater than 15 mm; the second convex lens assembly comprises a third convex lens and a fourth convex lens; The wavelength of the second laser is 980 nanometers and the power is 10 watts; The second dynamic structure fluorescence generated inside the living body to be imaged is generated inside the living body to be imaged after the second light beam in the first parallel Gaussian light spot passes through the near-infrared structure light generating device (2) and then irradiates the inside of the living body to be imaged.

3. The three-dimensional in vivo imaging system according to claim 2, characterized in that: The near-infrared structured light generating device (2) is a near-infrared spatial light modulator; Among them, the pixel array size of the near-infrared spatial light modulator is 912×1140, the maximum structured light refresh frequency is 120Hz, and the applicable wavelength range of the near-infrared spatial light modulator is 700nm to 2500nm.

4. The three-dimensional in vivo imaging system according to claim 3, characterized in that: The first deformable mirror (3) is mounted on a five-axis translation platform; The first deformable mirror (3) has 69 silver mirrors with actuators; Each silver mirror can adjust its reflection angle, and its reflection angle is less than or equal to 15 degrees.

5. The three-dimensional in vivo imaging system according to claim 4, characterized in that: A near-infrared adaptive optical device (5), comprising a third convex lens component (51) for reducing the focus of a first light path, a first long-pass filter (52), and a wavefront sensor (53) for obtaining an image difference between the first light path and a specified light structure based on the first light path passing through the third convex lens component (51), the first long-pass filter (52) and the specified light structure in sequence; The wavelength range of the first long-pass filter (52) is 1000 nm to 1500 nm; wherein the wavefront sensor (53) is arranged on a three-axis displacement platform.

6. The three-dimensional in vivo imaging system according to claim 5, characterized in that: The imaging device (6) comprises: a second long-pass filter (61), an imaging lens (62), and a first near-infrared camera (63) which are arranged in sequence; The wavelength range of the second long pass filter (61) is 1000nm to 1500nm; The first near-infrared camera (63) is an InGaAs near-infrared camera with a resolution of 640×512 pixels.

7. The three-dimensional in vivo imaging system according to claim 6, characterized in that: All convex lenses have an optical size of 1 inch in diameter.

8. The three-dimensional in vivo imaging system according to claim 7, characterized in that: The living body to be imaged is pre-processed, wherein the pre-processing method of the living body to be imaged is: The first fluorescent probe is injected into the living body to be imaged through the tail vein or injected into the living body to be imaged through gavage; The first fluorescent probe is a fluorescent probe that can be excited by near-infrared lasers of 808 nanometers and 980 nanometers and has an emission wavelength between 1000 nanometers and 1700 nanometers.

9. The three-dimensional in vivo imaging system according to claim 8, characterized in that: A three-dimensional reconstruction device (7) uses MatLab image processing software to reconstruct the two-dimensional image corresponding to the living body to be imaged into a three-dimensional model; The three-dimensional reconstruction device (7) uses Alpao Core Engine software to control the first deformable mirror (3) according to the aberration between the first light path and the specified light structure, so as to reduce the aberration between the first light path and the specified light structure within a preset range.

10. A three-dimensional in vivo imaging method, characterized in that: The method is performed by the three-dimensional in vivo imaging system according to any one of claims 5 to 9, and the method comprises: S1, placing a standard chessboard positioning plate in the field of view of a first near-infrared camera (63), using a near-infrared structured light generating device (2) to generate a first dynamic structured light and obtaining a two-dimensional image of the standard chessboard positioning plate in the first near-infrared camera (63), and then calibrating a reference plane for three-dimensional imaging of a living body to be imaged in a three-dimensional reconstruction device (7); S2, placing the living body to be imaged on a calibrated reference plane, using the first parallel Gaussian light spot emitted by the excitation device (1), and then the near-infrared structured light generating device (2), modulating the first parallel Gaussian light spot into a first dynamic structured light with the same specified light structure, and irradiating the first dynamic structured light onto the living body to be imaged; S3, a first deformable mirror (3), used to reflect the structural fluorescence signal inside the living body to be imaged to the first dichroic mirror (4); the structural fluorescence signal inside the living body to be imaged includes the fluorescence generated inside the living body to be imaged and the first dynamic structured light reflected by the surface of the living body to be imaged after the first dynamic structured light is irradiated onto the living body to be imaged; The fluorescence generated inside the living body to be imaged includes: a first dynamic structure fluorescence and a second dynamic structure fluorescence generated when the first dynamic structure light is irradiated inside the living body to be imaged; A first dichroic mirror (4) is used to separate the structural fluorescence signal inside the living body to be imaged, which is reflected by the first deformable mirror (3), into a first light path that enters a near-infrared adaptive optical device (5) and a second light path that enters an imaging device (6); The first light path only includes the first dynamic structure fluorescence emitted from the living body to be imaged; the second light path only includes the second dynamic structure fluorescence generated from the living body to be imaged; A near-infrared adaptive optical device (5), used for obtaining the aberration between the first light path and the specified light structure according to the first light path and the specified light structure; An imaging device (6) is used to acquire a two-dimensional image corresponding to the living body to be imaged based on the second light path; A three-dimensional reconstruction device (7) is used to reconstruct a two-dimensional image corresponding to the living body to be imaged into a three-dimensional model, and is used to control the first deformable mirror (3) according to the aberration between the first light path and the specified light structure, so that the aberration between the first light path and the specified light structure is reduced to within a preset range.

Images