A short-wave infrared Raman and fluorescence imaging device for living animals and its implementation method

Abstract

The present invention relates to the field of medical imaging technology, and more specifically, to a short-wave infrared Raman and fluorescence imaging device for living animals and its implementation method. The device comprises an imaging cavity, a short-wave infrared excitation module, a Raman imaging and fluorescence imaging module, a sample carrier, and an anesthesia unit. The sample carrier is disposed in the imaging cavity, the short-wave infrared excitation module is disposed on the side wall of the Raman imaging and fluorescence imaging module in the imaging cavity, the Raman imaging and fluorescence imaging module is disposed on the inner top of the imaging cavity, the anesthesia unit is disposed on one side of the imaging cavity, and the anesthesia unit is connected to the imaging cavity via a pipeline. The device can effectively improve imaging sensitivity and resolution, solve the problems of signal interference, inaccurate imaging registration, and multimodal integration existing in the prior art, and realize efficient and accurate imaging of living small animals in different imaging modes. The present invention is mainly used in the field of short-wave infrared Raman and fluorescence imaging of living animals.

Description

Technical Field

[0001] The present invention relates to the field of medical imaging technology, and more particularly to a short-wave infrared Raman and fluorescence imaging device for a living animal and an implementation method thereof. Background Art

[0002] Current multimodal imaging technologies for live small animals face significant technical bottlenecks. Traditional fluorescence imaging is limited by photon absorption and scattering, resulting in insufficient detection depth. Furthermore, tissue autofluorescence and photon scattering significantly reduce the signal-to-noise ratio. Furthermore, spectral overlap among fluorescent probes further impairs multi-target detection capabilities. While Raman imaging offers the advantage of molecular specificity, its weak signal intensity and slow imaging speed make it difficult to meet the demands of dynamic in vivo imaging, highlighting the inherent contradiction between sensitivity, resolution, and specificity of a single imaging modality.

[0003] Existing technologies achieve multimodal integration through discrete detectors, but the system is bulky and miniaturization is limited. The mechanical positioning error of the dual detectors is easily affected by environmental interference, resulting in insufficient spatial registration accuracy. In addition, due to the large difference in excitation light source power and poor timing synchronization between fluorescence and short-wave infrared Raman imaging, the signal coordination efficiency is significantly reduced, and the advantages of multimodal fusion cannot be fully utilized. At the same time, the physiological movement of living bodies exacerbates the imaging challenges. The difference in frame rates of different modalities causes temporal and spatial data mismatch, and motion artifacts lead to increased errors in quantitative analysis. Strategies to increase imaging depth often come at the expense of resolution, resulting in irreconcilable performance contradictions. Existing improvement solutions such as optical tomography technology, algorithm compensation and dual-mode probes either sacrifice molecular specificity, rely on artificial labeling and are inefficient, or cause biological toxicity. They have failed to fundamentally solve the core defects of multimodal coordination, system integration and dynamic imaging accuracy. Summary of the Invention

[0004] To overcome the shortcomings of the aforementioned prior art, the present invention provides a short-wave infrared Raman and fluorescence imaging device for living animals and its implementation method. This method effectively improves imaging sensitivity and resolution, addresses signal interference, inaccurate imaging registration, and multimodal integration challenges encountered in prior art, and enables efficient and accurate imaging of living small animals across various imaging modes.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0006] A shortwave infrared Raman and fluorescence imaging device for living animals comprises an imaging cavity, a shortwave infrared excitation module, a Raman imaging and fluorescence imaging module, a sample carrier and an anesthesia unit, wherein the sample carrier is arranged in the imaging cavity, the shortwave infrared excitation module is arranged on the side wall of the Raman imaging and fluorescence imaging module in the imaging cavity, the Raman imaging and fluorescence imaging module is arranged on the inner top of the imaging cavity, the shortwave infrared excitation module and the Raman imaging and fluorescence imaging module are all aligned with the sample carrier, and the anesthesia unit is arranged on one side of the imaging cavity, and the anesthesia unit is connected to the imaging cavity through a pipeline.

[0007] The short-wave infrared excitation module includes a short-wave infrared light source, a laser adjustment device, a beam shaping system and a short-wave infrared optical filter. The short-wave infrared light source has an output wavelength range of 800-1200nm and is used to excite the Raman signal of the sample. The laser adjustment device includes a laser power regulator for adjusting the output intensity of the laser. The beam shaping system includes an optical lens and a reflector for adjusting the shape and spot size of the laser beam. The short-wave infrared optical filter is used to filter out stray light that may interfere with the Raman signal.

[0008] The Raman imaging and fluorescence imaging module includes a Raman and fluorescence detection system and a multimodal signal fusion system. The Raman and fluorescence detection system uses a near-infrared CCD detection module to collect Raman and fluorescence signals generated after excitation by a short-wave infrared light source. The multimodal signal fusion system fuses the Raman imaging and fluorescence imaging signals in time or space to form a comprehensive image.

[0009] The sample carrier is provided with a temperature control unit to maintain a constant temperature of the sample during the experiment. The sample carrier is precisely adjusted in the X, Y, and Z axis directions to ensure that the sample can be accurately positioned at the laser focus.

[0010] The anesthesia unit includes an anesthetic gas supply system, an anesthesia mask and ventilation system, and a real-time monitoring system. The anesthetic gas supply system includes an anesthetic gas supplier and a gas mixer. The anesthetic gas supplier and gas mixer accurately control the concentration and flow of the anesthetic gas. The anesthesia mask and ventilation system provide gas ventilation support for the animal. The real-time monitoring system monitors the physiological parameters of the small animal's respiratory rate, body temperature and heart rate.

[0011] An imaging method for a short-wave infrared Raman and fluorescence imaging device for a living animal, characterized by comprising the following steps:

[0012] S1, the short-wave infrared excitation module emits short-wave infrared laser, irradiates the surface of the animal or sample, and stimulates the Raman signal of the living animal by irradiating the short-wave infrared light;

[0013] S2, Raman imaging and fluorescence imaging modules receive Raman signals and fluorescence signals respectively. After optical filtering and detection, the signals are reconstructed and processed by the computer system for multimodal fusion.

[0014] S3, the sample stage can accurately position and adjust the small animal according to experimental requirements, ensuring that the sample is always in the focus area during the imaging process;

[0015] S4, anesthesia unit provides stable anesthesia support to ensure the quietness and stability of small animals during the entire imaging process.

[0016] In step S1, a short-wave infrared Raman laser and a fluorescence excitation light are emitted respectively by a short-wave infrared excitation module. In the Raman imaging mode, the short-wave infrared laser excites the small animal to generate a Raman scattering signal; in the fluorescence imaging mode, the fluorescence excitation light source causes the small animal to emit a fluorescence signal.

[0017] In step S2, the excited Raman signal and the fluorescence signal are guided to the same near-infrared CCD detection module through optical elements. The near-infrared CCD detection module captures the Raman or fluorescence signal in real time according to the switching of the current imaging mode. The captured signal is amplified and filtered by the signal processing module. After filtering out the noise signal, the Raman and fluorescence image information are fused. The fused imaging results are displayed in real time through the control system, and a visual analysis function is provided to help researchers simultaneously obtain Raman imaging and fluorescence imaging data of small animals in the same field of view, thereby improving the overall imaging effect.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] By adopting time series switching technology, the system can accurately collect signals under different imaging modes, avoiding interference between the two signals. This precise control improves the clarity and accuracy of the signal, thereby improving the sensitivity and resolution of the overall imaging, especially in the imaging of living small animals, more subtle biological information can be clearly observed; by performing multimodal integration under a single detection module, and combining time series switching and wavelength selection technology, the present invention avoids the problems of signal interference and registration errors in different imaging modes. The two imaging modes can be efficiently fused to ensure the accurate alignment of Raman and fluorescence image data, eliminating the errors caused by signal fusion in the existing technology; short-wave infrared Raman imaging technology has good tissue penetration and is particularly suitable for imaging deep structures in small animals. By combining with fluorescence imaging, the present invention can achieve accurate imaging of different depths in small animals, broadening the depth and breadth of research; by combining Raman and fluorescence imaging modes, researchers can obtain more comprehensive biological information in the same experiment. This multimodal imaging method can simultaneously provide molecular specificity and high-resolution imaging data, helping to gain a deeper understanding of the dynamic changes in cells and tissues and promoting the advancement of life science research. By utilizing a single detection module, the device simplifies the overall structure of the device and reduces hardware complexity. Through time series switching and wavelength selection, the system achieves seamless switching between different imaging modes, simplifying the operation process, improving the system's operability and efficiency, and enabling high-precision multimodal imaging without damaging small animals. By reducing interference in animal experiments, the reliability of experimental data is improved, providing a more precise experimental tool for biomedical research. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is a schematic diagram of the structure of the present invention;

[0021] In the figure: 1 is the short-wave infrared excitation module, 2 is the Raman imaging and fluorescence imaging module, 3 is the sample carrier, and 4 is the anesthesia unit. DETAILED DESCRIPTION

[0022] In order to more clearly understand the above-mentioned objects, features and advantages of the present invention, the present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments. It should be noted that, in the absence of conflict, the embodiments of the present application and the features therein can be combined with each other.

[0023] In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention may also be implemented in other ways different from the description. Therefore, the scope of protection of the present invention is not limited to the specific embodiments disclosed below.

[0024] like Figure 1As shown, a short-wave infrared Raman and fluorescence imaging device for in-vivo animals includes an imaging chamber, a short-wave infrared excitation module 1, a Raman imaging and fluorescence imaging module 2, a sample carrier 3, and an anesthesia unit 4. The sample carrier 3 is positioned within the imaging chamber, the short-wave infrared excitation module 1 is positioned on a side wall of the imaging chamber, and the Raman imaging and fluorescence imaging module 2 is positioned at the inner ceiling of the imaging chamber. Both the short-wave infrared excitation module 1 and the Raman imaging and fluorescence imaging module 2 are aligned with the sample carrier 3. The anesthesia unit 4 is positioned on one side of the imaging chamber and connected to the imaging chamber via a pipeline. The short-wave infrared excitation module 1 provides short-wave infrared laser and fluorescence excitation light sources. The short-wave infrared laser source is used for short-wave infrared Raman imaging, while the fluorescence excitation light source is used for fluorescence imaging. The laser source must meet the excitation wavelength requirements of both imaging modes and ensure that the wavelength selection does not interfere with each other. The device is compatible with both Raman and fluorescence imaging. The module has a built-in high-sensitivity detector that can capture Raman scattering signals and fluorescence signals respectively. The system performs time-series switching or wavelength selection of signals in both imaging modes, and avoids interference between the two modes through appropriate signal filtering and modulation. This design reduces the registration error and signal fusion problems in multimodal integration through a unified detection module, while ensuring high-sensitivity and high-resolution imaging effects.

[0025] Preferably, the short-wave infrared excitation module 1 includes a short-wave infrared light source, a laser adjustment device, a beam shaping system and a short-wave infrared optical filter. The short-wave infrared light source has an output wavelength range of 800-1200nm, which is used to excite the Raman signal of the sample. The laser adjustment device includes a laser power regulator for adjusting the output intensity of the laser. The beam shaping system includes an optical lens and a reflector for adjusting the shape and spot size of the laser beam to ensure that the laser can be evenly irradiated on the sample. The short-wave infrared optical filter is used to filter out stray light that may interfere with the Raman signal to ensure the purity of the excitation light.

[0026] Preferably, the Raman and fluorescence imaging module 2 includes a Raman and fluorescence detection system and a multimodal signal fusion system. The Raman and fluorescence detection system uses a near-infrared CCD detection module to collect Raman and fluorescence signals generated after excitation by a short-wave infrared light source. The multimodal signal fusion system temporally or spatially fuses the Raman and fluorescence imaging signals to form a composite image. The Raman and fluorescence imaging module 2 is located at the top center of the device and uses a vertical optical path for signal acquisition and multimodal imaging.

[0027] Preferably, the sample carrier 3 is equipped with a temperature control unit to maintain a constant sample temperature during the experiment. The sample carrier 3 can be precisely adjusted in the X, Y, and Z axes to ensure that the sample can be accurately positioned at the laser focus. The sample carrier 3 uses a high-precision mechanical structure design to reduce sample vibration and ensure image clarity and stability during imaging.

[0028] Preferably, the anesthesia unit includes an anesthetic gas supply system, an anesthesia mask and ventilation system, and a real-time monitoring system. The anesthetic gas supply system includes an anesthetic gas supplier and a gas mixer. The anesthetic gas supplier and the gas mixer accurately control the concentration and flow of the anesthetic gas. The anesthesia mask and ventilation system provide gas ventilation support for the animal. The real-time monitoring system monitors the physiological parameters of the small animal's respiratory rate, body temperature and heart rate.

[0029] An imaging method for a short-wave infrared Raman and fluorescence imaging device for a living animal, characterized by comprising the following steps:

[0030] S1, the short-wave infrared excitation module 1 emits a short-wave infrared laser, irradiates the surface of the animal or sample, and stimulates the Raman signal of the living animal by irradiating the short-wave infrared light;

[0031] S2, Raman imaging and fluorescence imaging module 2 receive Raman signals and fluorescence signals respectively. After the signals are optically filtered and detected, the computer system performs image reconstruction and multimodal fusion processing;

[0032] S3, sample stage 3 can accurately position and adjust the small animal according to experimental requirements, ensuring that the sample is always in the focus area during the imaging process;

[0033] S4, anesthesia unit 4 provides stable anesthesia support to ensure the quietness and stability of the small animals during the entire imaging process.

[0034] Preferably, in step S1, the short-wave infrared excitation module 1 emits short-wave infrared Raman laser light and fluorescence excitation light, respectively. In Raman imaging mode, the short-wave infrared laser excites the small animal to produce a Raman scattering signal; in fluorescence imaging mode, the fluorescence excitation light source causes the small animal to emit a fluorescence signal. This design avoids interference between different detection channels and makes signal processing more efficient.

[0035] Preferably, in step S2, the excited Raman signal and the fluorescence signal are guided to the same near-infrared CCD detection module through optical elements. The near-infrared CCD detection module captures the Raman or fluorescence signal in real time according to the switching of the current imaging mode. The captured signal is amplified and filtered by the signal processing module. After filtering out the noise signal, the Raman and fluorescence image information are fused. The image registration and signal enhancement in this process are the key technologies of the present invention. The image is accurately processed by a special algorithm so that the data of the two imaging modes can be accurately aligned in space, thereby realizing efficient registration of multimodal imaging. The fused imaging results are displayed in real time through the control system, and a visual analysis function is provided to help researchers simultaneously obtain Raman imaging and fluorescence imaging data of small animals in the same field of view, thereby improving the comprehensive imaging effect.

[0036] This method effectively addresses the limitations of current small animal in vivo imaging technology, particularly in sensitivity, resolution, and deep penetration. By integrating fluorescence imaging with short-wave infrared Raman imaging, the present invention can provide more accurate imaging data and effectively overcome issues such as signal interference and registration errors in existing technologies, providing a new technical approach for small animal in vivo imaging research.

[0037] Based on the unique deep tissue penetration advantage of the shortwave infrared band, this technology is particularly suitable for the visualization study of deep biological structures in living small animal models. By integrating the complementary advantages of two imaging modalities, the system achieves multi-level precise observation from the epidermis to deep tissues, significantly expanding the spatial and information dimensions of biomedical research. The device uses advanced multi-spectral analysis technology to simultaneously collect Raman spectral features and fluorescence signal data in a single experiment. This multimodal joint application can not only synchronously output molecular fingerprint information and fine morphological features, but also reveal the dynamic behavior mechanism of the cell microenvironment through data fusion technology, providing a new observation perspective for life science research. The specially designed integrated detection unit integrates the traditional discrete detection system into a compact module through optical path optimization design, effectively streamlining the system architecture and reducing maintenance costs.

[0038] The above only describes in detail the preferred embodiments of the present invention, but the present invention is not limited to the above embodiments. Various changes can be made within the scope of knowledge possessed by ordinary technicians in this field without departing from the purpose of the present invention. Various changes should be included in the scope of protection of the present invention.

Claims

1. A short-wave infrared Raman and fluorescence imaging device for living animals, characterized by: The invention comprises an imaging cavity, a short-wave infrared excitation module (1), a Raman imaging and fluorescence imaging module (2), a sample carrier (3) and an anesthesia unit (4), wherein the sample carrier (3) is arranged in the imaging cavity, the short-wave infrared excitation module (1) is arranged on the side wall of the imaging cavity, the Raman imaging and fluorescence imaging module (2) is arranged on the inner top of the imaging cavity, the short-wave infrared excitation module (1) and the Raman imaging and fluorescence imaging module (2) are both aligned with the sample carrier (3), and the anesthesia unit (4) is arranged on one side of the imaging cavity, and the anesthesia unit (4) is connected to the imaging cavity through a pipeline; The short-wave infrared excitation module (1) comprises a short-wave infrared light source, a laser adjustment device, a beam shaping system and a short-wave infrared optical filter. The short-wave infrared light source has an output wavelength range of 800-1200 nm and is used to excite the Raman signal of the sample. The laser adjustment device comprises a laser power regulator for adjusting the output intensity of the laser. The beam shaping system comprises an optical lens and a reflector for adjusting the shape and spot size of the laser beam. The short-wave infrared optical filter is used to filter out stray light that may interfere with the Raman signal. The Raman imaging and fluorescence imaging module (2) includes a Raman and fluorescence detection system and a multimodal signal fusion system. The Raman and fluorescence detection system uses a near-infrared CCD detection module to collect Raman and fluorescence signals generated after excitation by a short-wave infrared light source. The multimodal signal fusion system fuses the Raman imaging and fluorescence imaging signals in time or space to form a comprehensive image.

2. The short-wave infrared Raman and fluorescence imaging device for living animals according to claim 1, characterized in that: The sample carrier (3) is provided with a temperature control unit to maintain a constant temperature of the sample during the experiment. The sample carrier (3) is precisely adjusted in the X, Y, and Z axis directions to ensure that the sample can be accurately positioned at the laser focus.

3. The short-wave infrared Raman and fluorescence imaging device for living animals according to claim 1, characterized in that: The anesthesia unit includes an anesthetic gas supply system, an anesthesia mask and ventilation system, and a real-time monitoring system. The anesthetic gas supply system includes an anesthetic gas supplier and a gas mixer. The anesthetic gas supplier and gas mixer accurately control the concentration and flow of the anesthetic gas. The anesthesia mask and ventilation system provide gas ventilation support for the animal. The real-time monitoring system monitors the physiological parameters of the small animal's respiratory rate, body temperature and heart rate.

4. An imaging method for the short-wave infrared Raman and fluorescence imaging device for living animals as claimed in claim 1, characterized in that: The following steps are involved: S1, short-wave infrared excitation module (1) emits short-wave infrared laser, irradiates the surface of the animal, and stimulates the Raman signal of the living animal by irradiating the short-wave infrared light; S2, Raman imaging and fluorescence imaging modules (2) receive Raman signals and fluorescence signals respectively. After the signals are optically filtered and detected, the computer system performs image reconstruction and multimodal fusion processing; S3, the sample stage (3) can accurately position and adjust the small animal according to the experimental requirements, ensuring that the sample is always in the focus area during the imaging process; S4, anesthesia unit (4) provides stable anesthesia support to ensure that the small animals are quiet and stable throughout the imaging process.

5. The method for short-wave infrared Raman and fluorescence imaging of living animals according to claim 4, characterized in that: In step S1, the short-wave infrared excitation module (1) emits short-wave infrared Raman laser and fluorescence excitation light respectively. In the Raman imaging mode, the short-wave infrared laser excites the small animal to generate a Raman scattering signal; in the fluorescence imaging mode, the fluorescence excitation light causes the small animal to emit a fluorescence signal.

6. The method for short-wave infrared Raman and fluorescence imaging of living animals according to claim 4, characterized in that: In step S2, the excited Raman signal and the fluorescence signal are guided to the same near-infrared CCD detection module through optical elements. The near-infrared CCD detection module captures the Raman or fluorescence signal in real time according to the switching of the current imaging mode. The captured signal is amplified and filtered by the signal processing module. After filtering out the noise signal, the Raman and fluorescence image information are fused. The fused imaging results are displayed in real time through the control system, and a visual analysis function is provided to help researchers simultaneously obtain Raman imaging and fluorescence imaging data of small animals in the same field of view, thereby improving the overall imaging effect.

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