Short-wave infrared Raman and fluorescence imaging device for living animals and implementation method of short-wave infrared Raman and fluorescence imaging device

Abstract

The invention relates to the technical field of medical imaging, in particular to a short-wave infrared Raman and fluorescence imaging device for an animal living body and an implementation method of the short-wave infrared Raman and fluorescence imaging device. Comprising an imaging cavity, a short-wave infrared excitation module, a Raman imaging and fluorescence imaging module, a sample carrying table and an anesthesia unit, the sample carrying table is arranged in the imaging cavity, the short-wave infrared excitation module is arranged on the side wall of the Raman imaging and fluorescence imaging module of the imaging cavity, and the Raman imaging and fluorescence imaging module is arranged at the inner top of the imaging cavity; the anesthesia unit is arranged on one side of the imaging cavity and communicated with the imaging cavity through a pipeline. According to the device, the imaging sensitivity and resolution can be effectively improved, the problems of signal interference, inaccurate imaging registration and multi-mode integration in the prior art are solved, and efficient and accurate imaging of small living animals in different imaging modes is realized. The method is mainly applied to 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 specifically, to an in-vivo short-wave infrared Raman and fluorescence imaging device for animals and a method for realizing the same. Background Art

[0002] At present, there are significant technical bottlenecks in the multi-modal imaging technology for small in-vivo animals. Traditional fluorescence imaging is limited by photon absorption and scattering, with insufficient detection depth. Moreover, autofluorescence of biological tissues and photon scattering lead to a significant decrease in the signal-to-noise ratio. At the same time, the spectral overlap of fluorescence probes further weakens the multi-target detection ability. Although Raman imaging has the advantage of molecular specificity, its signal intensity is weak and the imaging speed is low, making it difficult to meet the requirements of in-vivo dynamic imaging, highlighting the inherent contradiction among sensitivity, resolution, and specificity in a single imaging mode.

[0003] The prior art realizes multi-modal integration through discrete detectors, but the system is large in volume and limited in miniaturization. The mechanical positioning error of the dual detectors is easily interfered by the environment, resulting in insufficient spatial registration accuracy. In addition, due to the large difference in the power of the excitation light sources and poor timing synchronization between fluorescence and short-wave infrared Raman imaging, the signal cooperation efficiency is significantly reduced, and the advantages of multi-modal fusion cannot be fully utilized. At the same time, the in-vivo physiological movement exacerbates the imaging challenge. The frame rate differences in different modalities cause spatio-temporal data mismatch, and motion artifacts increase the quantitative analysis error; strategies for increasing the imaging depth often come at the cost of sacrificing resolution, forming irreconcilable performance contradictions. Existing improvement schemes 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, and none of them can fundamentally solve the core defects of multi-modal cooperation, system integration, and dynamic imaging accuracy. Summary of the Invention

[0004] To overcome the deficiencies in the above prior art, the present invention provides an in-vivo short-wave infrared Raman and fluorescence imaging device for animals and a method for realizing the same. This method can effectively improve the imaging sensitivity and resolution, solve the problems of signal interference, inaccurate imaging registration, and multi-modal integration existing in the prior art, and achieve efficient and accurate imaging of small in-vivo animals in different imaging modes.

[0005] To solve the above technical problems, the technical solution adopted by the present invention is as follows: An in-vivo animal short-wave infrared Raman and fluorescence imaging device, comprising an imaging chamber, a short-wave infrared excitation module, a Raman imaging and fluorescence imaging module, a sample stage and an anesthesia unit. The sample stage is arranged in the imaging chamber. The short-wave infrared excitation module is arranged on the side wall of the Raman imaging and fluorescence imaging module of the imaging chamber. The Raman imaging and fluorescence imaging module is arranged at the inner top of the imaging chamber. Both the short-wave infrared excitation module and the Raman imaging and fluorescence imaging module are arranged to be aligned with the sample stage. The anesthesia unit is arranged on one side of the imaging chamber, and the anesthesia unit is communicated with the imaging chamber through a pipeline.

[0006] 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 output wavelength range of the short-wave infrared laser source is generally 800 - 1200 nm, 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 optical lenses and mirrors 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.

[0007] 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 for detection, which is used to collect the Raman and fluorescence signals generated after short-wave infrared excitation. The multimodal signal fusion system fuses the Raman imaging and fluorescence imaging signals in time or space to form a comprehensive image.

[0008] A temperature control unit is arranged in the sample stage to maintain the constant temperature of the sample during the experiment. The sample stage is precisely adjusted in the X, Y, and Z axis directions to ensure that the sample can be accurately positioned at the laser focus.

[0009] The anesthesia unit includes an anesthesia gas supply system, an anesthesia mask and ventilation system, and a real-time monitoring system. The anesthesia gas supply system includes an anesthesia gas supplier and a gas mixer. The anesthesia gas supplier and the gas mixer precisely control the concentration and flow rate of the anesthesia gas. The anesthesia mask and ventilation system provides gas ventilation support for the animal. The real-time monitoring system monitors the physiological parameters such as the respiratory rate, body temperature, and heart rate of the small animal.

[0010] An imaging method applied to an in-vivo animal short-wave infrared Raman and fluorescence imaging device, characterized by comprising the following steps: S1. The short-wave infrared excitation module emits short-wave infrared laser, which covers the surface of the animal or sample, and excites the Raman signal of the sample by means of short-wave infrared light irradiation; S2. The Raman imaging and fluorescence imaging modules respectively receive Raman signals and fluorescence signals. After the signals are optically filtered and detected, the computer system performs image reconstruction and multimodal fusion processing; S3. The sample stage can accurately position and adjust small animals according to experimental requirements to ensure that the sample is always in the focal area during imaging; S4. The anesthesia unit provides stable anesthesia support to ensure the quietness and stability of small animals during the entire imaging process.

[0011] In step S1, the short-wave infrared excitation module emits short-wave infrared Raman laser and fluorescence excitation light source respectively. In the Raman imaging mode, the short-wave infrared laser excites the sample to generate Raman scattering signals; in the fluorescence imaging mode, the excitation light source causes the sample to emit fluorescence signals.

[0012] In step S2, the excited Raman signals and fluorescence signals are guided to the same detection module through optical elements. The detection module captures Raman or fluorescence signals in real time according to the switching of the current imaging mode. The captured signals are amplified and filtered by the signal processing module. After filtering out the noise signals, the Raman and fluorescence image information is fused. Through the control system, the fused imaging results are displayed in real time and a visualization analysis function is provided to help researchers obtain the Raman imaging and fluorescence imaging data of small animals in the same field of view simultaneously, improving the comprehensive effect of imaging.

[0013] Compared with the prior art, the beneficial effects of the present invention are: By adopting time - series switching technology, the system can accurately collect signals in different imaging modes, avoiding interference between the two signals. This precise control improves the clarity and accuracy of the signals, thereby enhancing the overall imaging sensitivity and resolution. Especially in the imaging of live small animals, more subtle biological information can be clearly observed. By integrating multiple modalities under a single detection module and combining time - series switching and wavelength - selection technology, the present invention avoids signal interference and registration error problems 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 prior art. The short - wave infrared Raman imaging technology has good tissue penetration and is particularly suitable for imaging deep - seated structures in small animals. By combining with fluorescence imaging, the present invention can achieve precise 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 can simultaneously provide molecular - specific and high - resolution imaging data, contributing to a deeper understanding of the dynamic changes of cells and tissues and promoting the progress of life - science research. The device simplifies the overall structure of the equipment by adopting a single detection module, reducing the complexity of the hardware. Through time - series switching and wavelength selection, the system realizes seamless switching between different imaging modes, simplifies the operation process, improves the operability and usage efficiency of the system, and can perform high - precision multimodal imaging without damaging small animals. By reducing interference in animal experiments, the reliability of experimental data is improved, providing a more accurate experimental tool for biomedical research. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is a schematic structural diagram of the present invention; In the figure: 1 is a short - wave infrared excitation module, 2 is a Raman imaging and fluorescence imaging module, 3 is a sample stage, and 4 is an anesthesia unit. DETAILED DESCRIPTION OF THE INVENTION

[0015] In order to more clearly understand the above - mentioned objects, features, and advantages of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments. It should be noted that, without conflict, the embodiments of the present application and the features in the embodiments can be combined with each other.

[0016] In the following description, many specific details are set forth in order to fully understand the present invention. However, the present invention can also be implemented in other ways different from those described herein. Therefore, the protection scope of the present invention is not limited by the specific embodiments disclosed below.

[0017] Such as Figure 1As shown in the figure, a short-wave infrared Raman and fluorescence imaging device for live animals includes an imaging chamber, a short-wave infrared excitation module 1, a Raman imaging and fluorescence imaging module 2, a sample stage 3, and an anesthesia unit 4. The sample stage 3 is arranged in the imaging chamber, the short-wave infrared excitation module 1 is arranged on the side wall of the imaging chamber, the Raman imaging and fluorescence imaging module 2 is arranged on the inner top of the imaging chamber, both the short-wave infrared excitation module 1 and the Raman imaging and fluorescence imaging module 2 are arranged to be aligned with the sample stage 3, the anesthesia unit 4 is arranged on one side of the imaging chamber, and the anesthesia unit 4 is communicated with the imaging chamber through a pipeline. The short-wave infrared excitation module 1 provides a short-wave infrared laser and a fluorescence excitation light source. The short-wave infrared laser light source is used for short-wave infrared Raman imaging, and the fluorescence excitation light source is used for fluorescence imaging. The selection of the laser source needs to meet the excitation wavelength requirements of the two imaging modes and ensure that the wavelength selection does not cause mutual interference. The device is compatible with both Raman and fluorescence imaging at the same time. This module is built with a highly sensitive detector, which can be used to capture Raman scattering signals and fluorescence signals respectively. The system switches the signals in a time series or selects wavelengths in the two imaging modes, and through appropriate signal filtering and modulation, it avoids interference between the two modes. This design reduces the registration error and signal fusion problem in multimodal integration through a unified detection module, and at the same time ensures high-sensitivity and high-resolution imaging effects.

[0018] 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 output wavelength range of the short-wave infrared laser source is usually 800 - 1200 nm, which is used to excite the Raman signal of the sample. The laser adjustment device includes a laser power regulator, which is used to adjust the output intensity of the laser. The beam shaping system includes optical lenses and mirrors, which are used to adjust the shape and spot size of the laser beam to ensure that the laser can uniformly irradiate the sample. The short-wave infrared optical filter is used to filter out the stray light that may interfere with the Raman signal and ensure the purity of the excitation light.

[0019] Preferably, 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 detector to collect the Raman and fluorescence signals generated after short-wave infrared excitation. The multimodal signal fusion system fuses the Raman imaging and fluorescence imaging signals in time or space to form a comprehensive image. The Raman imaging and fluorescence imaging module 2 is located in the center of the top of the device, and uses a vertical optical path to collect signals and realizes the multimodal imaging function.

[0020] Preferably, a temperature control unit is arranged in the sample stage 3 to maintain the constant temperature of the sample during the experiment. The sample stage 3. The sample stage 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. The sample stage 3 reduces sample vibration through a high-precision mechanical structure design to ensure the clarity and stability of the image during the imaging process.

[0021] Preferably, the anesthesia unit includes an anesthesia gas supply system, an anesthesia mask and ventilation system, and a real-time monitoring system. The anesthesia gas supply system includes an anesthesia gas supplier and a gas mixer. The anesthesia gas supplier and the gas mixer precisely control the concentration and flow rate of the anesthesia gas. The anesthesia mask and ventilation system provides gas ventilation support for the animal. The real-time monitoring system monitors the physiological parameters of the respiratory rate, body temperature, and heart rate of the small animal.

[0022] An imaging method applied to an in-vivo short-wave infrared Raman and fluorescence imaging device for animals, characterized by comprising the following steps: S1. The short-wave infrared excitation module 1 emits short-wave infrared laser, which covers the surface of the animal or sample, and excites the Raman signal of the sample by means of short-wave infrared light irradiation; S2. The Raman imaging and fluorescence imaging module 2 respectively receives the Raman signal and the fluorescence signal. 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 to ensure that the sample is always in the focal area during the imaging process; S4. The anesthesia unit 4 provides stable anesthesia support to ensure the quietness and stability of the small animal during the entire imaging process.

[0023] Preferably, in step S1, the short-wave infrared excitation module 1 respectively emits short-wave infrared Raman laser and fluorescence excitation light source. In the Raman imaging mode, the short-wave infrared laser excites the sample to generate Raman scattering signal; in the fluorescence imaging mode, the excitation light source makes the sample emit fluorescence signal. This design avoids interference between different detection channels and makes signal processing more efficient.

[0024] Preferably, in step S2, the excited Raman signal and fluorescence signal are guided to the same detection module through optical elements. The 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 is fused. Image registration and signal enhancement in this process are the key technologies of the present invention. The images are precisely processed by a dedicated algorithm, so that the data of the two imaging modes can be accurately aligned in space, thereby realizing efficient registration of multimodal imaging. Through the control system, the fused imaging result is displayed in real time and a visualization analysis function is provided to help researchers obtain the Raman imaging and fluorescence imaging data of small animals simultaneously in the same field of view, improving the comprehensive effect of imaging.

[0025] Effectively solve the limitations existing in current small animal in vivo imaging technology, especially the deficiencies in sensitivity, resolution, and deep penetration ability. Through the integration of fluorescence imaging and short-wave infrared Raman imaging, the present invention can provide more accurate imaging data and effectively overcome problems such as signal interference and registration errors in the prior art, providing a new technical means for small animal in vivo imaging research.

[0026] Based on the unique deep tissue penetration advantage of the short-wave infrared band, this technology is particularly suitable for the visualization research of deep biological structures in small animal models in vivo. By integrating the complementary advantages of the two imaging modalities, the system realizes multi-level precise observation from the epidermis to deep tissues, significantly expanding the spatial dimension and information dimension of biomedical research. The device adopts advanced multi-spectral analysis technology and can simultaneously collect Raman spectral features and fluorescence signal data in a single experiment. This joint application of multi-modalities 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, through optimized optical path design, integrates the traditional discrete detection systems into a compact module, effectively streamlining the system architecture and reducing maintenance costs.

[0027] Only the preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those of ordinary skill in the art, various changes can be made without departing from the gist of the present invention, and all such changes should be included within the protection scope of the present invention.

Claims

1. A short-wave infrared Raman and fluorescence imaging device for living animals, characterized in that: 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 arranged to be 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.

2. The short-wave infrared Raman and fluorescence imaging device for living animals according to claim 1, characterized in that: 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 laser source outputs a wavelength range of usually 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.

3. The short-wave infrared Raman and fluorescence imaging device for living animals according to claim 1, characterized in that: The Raman imaging and fluorescence imaging module (2) comprises a Raman and fluorescence detection system and a multimodal signal fusion system. The Raman and fluorescence detection system adopts near-infrared CCD detection to collect Raman and fluorescence signals generated after short-wave infrared excitation. The multimodal signal fusion system fuses the Raman imaging and fluorescence imaging signals in time or space to form a comprehensive image.

4. 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 directions of the X, Y and Z axes to ensure that the sample can be accurately positioned at the laser focus.

5. 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 anesthetic 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 anesthetic mask and ventilation system provide gas ventilation support for the animal. The real-time monitoring system monitors the physiological parameters of the respiratory rate, body temperature and heart rate of the small animal.

6. An imaging method applied to 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, a short-wave infrared excitation module (1) emits a short-wave infrared laser, which is placed on the surface of the animal or sample to excite the Raman signal of the sample by irradiating the short-wave infrared light; S2, Raman imaging and fluorescence imaging module (2) receive Raman signals and fluorescence signals respectively, and after the signals are optically filtered and detected, the computer system performs image reconstruction and multi-modal 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.

7. The method for short-wave infrared Raman and fluorescence imaging of living animals according to claim 6, characterized in that: In step S1, a short-wave infrared excitation module (1) emits a short-wave infrared Raman laser and a fluorescence excitation light source respectively. In the Raman imaging mode, the short-wave infrared laser excites the sample to generate a Raman scattering signal; in the fluorescence imaging mode, the excitation light source causes the sample to emit a fluorescence signal.

8. The method for short-wave infrared Raman and fluorescence imaging of living animals according to claim 6, characterized in that: In step S2, the excited Raman signal and the fluorescence signal are guided to the same detection module through optical elements. The 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 the Raman imaging and fluorescence imaging data of small animals in the same field of view, thereby improving the comprehensive imaging effect.

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