CN104181142A - Molecular image imaging verification system and method - Google Patents
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Abstract
Description
技术领域technical field
本发明涉及分子成像技术领域，尤其涉及一种结合冰冻切片机及CCD相机的分子影像成像验证系统和方法The invention relates to the technical field of molecular imaging, in particular to a molecular imaging verification system and method combined with a cryostat and a CCD camera
背景技术Background technique
随着基因组学、蛋白组学和疾病基因组学的迅速发展，疾病的诊断正在从传统的疾病表征观察、常规的生化实验检测，发展到多种基因和分子水平的微观特征认识，其中利用分子影成像技术可以从基因、蛋白质水平深刻认识疾病的发生、发展过程，能够实现现有微观分析所无法取代的整体、连续、无创的特异检测方法，生物在体分子成像理论及其技术将会提供全新的预防、诊断和治疗手段。与传统的医学影成像技术相比较而言，分子成像学着眼于构成疾病或病变的基础变化和基因分子水平的异常，而不是对由基因分子改变所构成的最终结果进行成像。在特异的分子探针的帮助下，分子成像技术可以在细胞、基因和分子水平上实现生物体内部生理或病理过程的无创实时动态在体成像，从而为疾病相关基因功能定位、细胞生长发育和突变过程的作用机制、新药研发等研究提供详细的定性、定位、定量资料以及有效的信息获取和分析处理的手段。With the rapid development of genomics, proteomics and disease genomics, the diagnosis of diseases is developing from the traditional observation of disease characterization and routine biochemical experiment detection to the recognition of microscopic features at the gene and molecular levels. Imaging technology can deeply understand the occurrence and development process of diseases from the level of genes and proteins, and can realize an integral, continuous, non-invasive specific detection method that cannot be replaced by existing microscopic analysis. The theory and technology of biological molecular imaging in vivo will provide a new prevention, diagnosis and treatment. Compared with traditional medical imaging technology, molecular imaging focuses on the basic changes and gene molecular level abnormalities that constitute diseases or lesions, rather than imaging the final results of gene molecular changes. With the help of specific molecular probes, molecular imaging technology can realize non-invasive real-time dynamic in vivo imaging of physiological or pathological processes inside organisms at the cellular, gene and molecular levels, so as to provide information on disease-related gene function localization, cell growth and development and Research on the mechanism of mutation process, new drug development and other research provides detailed qualitative, positioning, quantitative data and effective means of information acquisition and analysis and processing.
激发荧光成像的原理可以描述为：当外源光照射到带有荧光团的生物组织上时，荧光团吸收光能使得电子跃迁到了激发态，电子从激发态回到基态的过程中会释放出荧光，该荧光较吸收的光向红端移动，即发射的荧光比吸收的外源光的能量低，荧光在组织体内传播并有一部分达到体表，从体表发出的荧光被探测器接收到，从而形成荧光图像。一般而言，荧光团发射出的荧光经过组织体散射，光的强度已经很弱，用肉眼很难观测到，因此需要在完全避光的暗箱中进行成像，并且要求探测器的灵敏度要高，通常利用一个低温制冷的高度灵敏的CCD相机来探测组织体表的荧光光子。CCD相机的另一个优势是空间分辨率较高。The principle of excited fluorescence imaging can be described as: when external light is irradiated on the biological tissue with fluorophore, the fluorophore absorbs the light energy and makes electrons transition to the excited state, and the electrons will be released during the process of returning from the excited state to the ground state. Fluorescence, the fluorescence moves to the red end compared with the absorbed light, that is, the emitted fluorescence has lower energy than the absorbed external light, the fluorescence propagates in the tissue body and part of it reaches the body surface, and the fluorescence emitted from the body surface is received by the detector , forming a fluorescence image. Generally speaking, the fluorescence emitted by the fluorophore is scattered by the tissue body, and the intensity of the light is already very weak, which is difficult to observe with the naked eye. Therefore, imaging needs to be carried out in a dark box completely protected from light, and the sensitivity of the detector is required to be high. Typically, a cryogenically cooled, highly sensitive CCD camera is used to detect fluorescent photons from the tissue surface. Another advantage of CCD cameras is the higher spatial resolution.
由于光学分子成像固有的病态性及测量噪声等因素，成像结果通常与真实结果有一些差异。由于产生荧光的生物组织通常属于软组织，在一些结构成像模态(如CT、MRI等)中无法获得该生物组织的真实几何信息，难以提供真实可靠的荧光光源位置，使得评价光学分子成像方法的优劣性变得十分困难。Due to the inherent pathological nature of optical molecular imaging and measurement noise and other factors, the imaging results usually have some differences from the real results. Since the biological tissue that produces fluorescence usually belongs to soft tissue, the real geometric information of the biological tissue cannot be obtained in some structural imaging modalities (such as CT, MRI, etc.), and it is difficult to provide a true and reliable position of the fluorescent light source, making the evaluation of optical molecular imaging methods difficult. Pros and cons become very difficult.
发明内容Contents of the invention
本发明的目的是针对现有技术的缺陷，提供一种分子影像成像验证系统和方法，可以对待检测物体的横截面切片测量，大大简化了操作步骤和操作流程，系统结构合理，功能显著，操作方便。The purpose of the present invention is to address the defects of the prior art, to provide a molecular image imaging verification system and method, which can measure the cross-sectional slice of the object to be detected, greatly simplify the operation steps and operation process, the system structure is reasonable, the function is remarkable, and the operation convenient.
为实现上述目的，本发明提供了一种分子影像成像验证系统，所述系统包括图像采集部分与图像处理部分：To achieve the above object, the present invention provides a molecular imaging verification system, the system includes an image acquisition part and an image processing part:
所述图像采集部分包括冰冻切片机(1)、相机(2)、相机镜头(3)、采集装置支架(4)、相机支架(5)、相机滑动装置(6)、发射滤光片支架(7)、多个发射滤光片(8)、光纤(9)、激发光源(10)；The image acquisition part includes a cryostat (1), a camera (2), a camera lens (3), an acquisition device support (4), a camera support (5), a camera sliding device (6), an emission filter support ( 7), multiple emission filters (8), optical fibers (9), excitation light source (10);
所述冰冻切片机(1)的柜体与所述采集装置支架(4)相连接，所述采集装置支架(4)与所述相机滑动装置(6)相连接，所述相机滑动装置(6)与所述相机支架(5)连接，所述相机支架(5)与所述相机(2)连接；The cabinet of the frozen microtome (1) is connected with the acquisition device support (4), the acquisition device support (4) is connected with the camera sliding device (6), and the camera sliding device (6 ) is connected with the camera bracket (5), and the camera bracket (5) is connected with the camera (2);
所述相机(2)的转接口与所述相机镜头(3)的转接口相连接，所述相机镜头(3)的进光口与所述发射滤光片支架(7)相连接，所述发射滤光片(8)内嵌于所述发射滤光片支架(7)的卡槽中；The adapter of the camera (2) is connected with the adapter of the camera lens (3), the light inlet of the camera lens (3) is connected with the emission filter holder (7), and the The emission filter (8) is embedded in the slot of the emission filter holder (7);
所述激发光源(10)出口连接所述光纤(9)的一端，所述光纤(9)的另一端指向被观测物体；The outlet of the excitation light source (10) is connected to one end of the optical fiber (9), and the other end of the optical fiber (9) points to the object to be observed;
所述图像处理部分包括图像处理系统，与所述相机(2)连接，用于接收白光图像及激发荧光图像，并对荧光图像自动分割、伪彩色添加、自动分割区域的光子数统计处理、自动分割区域的几何信息测量，将荧光图像同白光图像叠加后获得激发荧光成像区域的真实几何位置。The image processing part includes an image processing system connected to the camera (2) for receiving white light images and exciting fluorescence images, and automatically segmenting the fluorescence images, adding false colors, automatically dividing the photon count of the region, and automatically The geometric information of the segmented area is measured, and the real geometric position of the excited fluorescence imaging area is obtained after the fluorescence image is superimposed on the white light image.
进一步的，所述图像处理系统包括：前处理模块(21)、分析模块(22)和存储模块(23)；Further, the image processing system includes: a pre-processing module (21), an analysis module (22) and a storage module (23);
所述前处理模块(21)与所述相机(2)的数据输出端口相连接，用于对接收到的荧光图像进行荧光强度均匀校正处理与自体荧光干扰去除处理，而对接收到的白光图像不做处理；The pre-processing module (21) is connected to the data output port of the camera (2), and is used to perform fluorescence intensity uniformity correction processing and autofluorescence interference removal processing on the received fluorescence image, and the received white light image do not deal with;
所述分析模块(22)与所述前处理模块(21)相连接，用于对前处理模块(21)发送的荧光图像依次进行自动分割、伪彩色添加、自动分割区域的光子数统计、自动分割区域的几何信息测量、与白光图像叠加的操作，并显示荧光图像经过自动分割、伪彩色添加、自动分割区域的光子数统计并与白光图像叠加后得到的图像；The analysis module (22) is connected to the pre-processing module (21), and is used to sequentially perform automatic segmentation, pseudo-color addition, photon number statistics of the automatic segmentation area, and automatic The geometric information measurement of the segmented area, the operation of superimposing with the white light image, and displaying the image obtained after the fluorescence image is automatically segmented, pseudo-color added, photon number statistics of the automatically segmented area and superimposed with the white light image;
所述存储模块(23)与所述前处理模块(21)和所述分析模块(22)连接，用于对前处理模块(21)处理后的荧光图像和白光图像进行保存，并对分析模块(22)处理后的，在荧光图像经过自动分割、伪彩色添加、自动分割区域的光子数统计并与白光图像叠加后得到的图像进行保存。The storage module (23) is connected to the pre-processing module (21) and the analysis module (22), and is used to save the fluorescence image and the white light image processed by the pre-processing module (21), and to store the pre-processing module (21) and the analysis module (22) After processing, the fluorescent image is saved after automatic segmentation, pseudo-color addition, photon number statistics of the automatically segmented area, and superimposition with the white light image.
进一步的，所述相机(2)是光电探测器。Further, the camera (2) is a photodetector.
为实现上述目的，本发明提供了一种基于上述分子影像成像验证系统的验证方法，其特征在于，所述方法包括：To achieve the above object, the present invention provides a verification method based on the above molecular imaging verification system, characterized in that the method includes:
步骤S1：通过控制相机滑动装置及相机支架的位置，使相机镜头的测量区域准确对应被检测切片，相机镜头视野完整包括被检测切片横截面区域，实现相机对被检测切片的清晰成像；Step S1: By controlling the position of the camera sliding device and the camera bracket, the measurement area of the camera lens accurately corresponds to the detected slice, and the field of view of the camera lens completely includes the cross-sectional area of the detected slice, so that the camera can clearly image the detected slice;
步骤S2：开启冰冻切片机白光灯，并控制相机获取白光图像，白光图像反应被检测切片的横截面信息；Step S2: Turn on the white light lamp of the cryostat, and control the camera to acquire a white light image, which reflects the cross-sectional information of the detected slice;
步骤S3：将发射滤光片放入发射滤光片支架，并连接发射滤光片支架和相机镜头；开启激发光源，同时关闭白光灯；控制相机获取荧光图像；关闭激发光源；荧光图像反应被检测切片包含的荧光光源的分布信息；Step S3: put the emission filter into the emission filter holder, and connect the emission filter holder and the camera lens; turn on the excitation light source, and turn off the white light at the same time; control the camera to acquire fluorescence images; turn off the excitation light source; Detect the distribution information of the fluorescent light source contained in the slice;
步骤S4：荧光图像和白光图像传送到图像处理部分的前处理模块；前处理模块对荧光图像进行强度校正操作和自体荧光去除处理操作；Step S4: The fluorescence image and the white light image are sent to the pre-processing module of the image processing part; the pre-processing module performs intensity correction and autofluorescence removal processing operations on the fluorescence image;
步骤S5：分析模块对前处理模块发送的荧光图像依次进行自动分割、伪彩色添加、自动分割区域的光子数统计、自动分割区域的几何信息测量、与白光图像叠加处理，并显示荧光图像经过自动分割、伪彩色添加、自动分割区域的光子数统计并与白光图像叠加后得到的图像；Step S5: The analysis module sequentially performs automatic segmentation, pseudo-color addition, photon number statistics of the automatically segmented area, geometric information measurement of the automatically segmented area, and superposition processing with the white light image on the fluorescence image sent by the pre-processing module, and displays the fluorescence image after automatic Segmentation, pseudo-color addition, photon number statistics of the automatically segmented area and the image obtained after superimposing with the white light image;
步骤S6：存储模块对前处理模块处理后的荧光图像和白光图像进行保存，并对分析模块处理后的，即在荧光图像经过自动分割、伪彩色添加、自动分割区域的光子数统计并与白光图像叠加后得到的图像进行保存，并保存测量得到的荧光区域几何信息。Step S6: The storage module saves the fluorescent image and the white light image processed by the pre-processing module, and counts the number of photons of the fluorescent image processed by the analysis module, that is, after automatic segmentation, pseudo-color addition, and automatic segmentation, and compares it with the white light image. The image obtained after image superimposition is saved, and the geometric information of the measured fluorescence area is saved.
本发明的有益效果是：建立了一套结合冰冻切片机及CCD相机的分子影像成像验证系统及验证方法。该系统及方法能够完成对小动物等待检测物体的横截面切片测量，在设定参数后可自动获取清晰的小动物切片荧光数据，完成白光图像采集、荧光图像采集及与白光图像叠加、荧光图像自动分割、分割区域光子数统计、荧光区域几何信息测量等处理，大大简化了操作步骤和操作流程，为分子影像成像结果提供金标准。本发明系统结构合理，功能显著，操作方便，可广泛应用于光学分子成像领域，具有广阔的市场前景。The beneficial effect of the invention is that a set of molecular image imaging verification system and verification method combined with a cryostat and a CCD camera is established. The system and method can complete the measurement of cross-sectional slices of small animals waiting to be detected, and can automatically obtain clear fluorescence data of small animal slices after setting parameters, and complete white light image acquisition, fluorescence image acquisition and superimposition with white light images, and fluorescence image acquisition. Automatic segmentation, statistics of photon counts in segmented areas, and geometric information measurement of fluorescent areas greatly simplifies the operation steps and procedures, providing the gold standard for molecular imaging results. The system of the invention has reasonable structure, significant functions and convenient operation, can be widely used in the field of optical molecular imaging, and has broad market prospects.
附图说明Description of drawings
图1为本发明分子影像成像验证系统的示意图；1 is a schematic diagram of a molecular imaging verification system of the present invention;
图2为本发明分子影像成像验证方法的流程图。Fig. 2 is a flow chart of the molecular imaging verification method of the present invention.
具体实施方式Detailed ways
下面通过附图和实施例，对本发明的技术方案做进一步的详细描述。The technical solutions of the present invention will be described in further detail below with reference to the accompanying drawings and embodiments.
图1为本发明分子影像成像验证系统的示意图，如图所示，本发明的系统包括图像采集部分与图像处理部分；图像采集部分包括冰冻切片机1、相机2、相机镜头3、采集装置支架4、相机支架5、相机滑动装置6、发射滤光片支架7、多个发射滤光片8、光纤9、激发光源10。Fig. 1 is the schematic diagram of the molecular image imaging verification system of the present invention, as shown in the figure, the system of the present invention includes an image acquisition part and an image processing part; the image acquisition part includes a cryostat 1, a camera 2, a camera lens 3, and an acquisition device bracket 4. Camera support 5 , camera sliding device 6 , emission filter support 7 , multiple emission filters 8 , optical fiber 9 , excitation light source 10 .
冰冻切片机1的柜体同采集装置支架4连接，采集装置支架4同相机滑动装置6连接，相机滑动装置6同相机支架5连接，相机支架5同相机2连接；相机2的转接口同相机镜头3的转接口连接，相机镜头3的进光口同发射滤光片支架7连接，发射滤光片8内嵌于发射滤光片支架7的卡槽中；激发光源10出口连接光纤9的一端，光纤9的另一端指向被观测物体。The cabinet of the cryostat 1 is connected with the acquisition device support 4, the acquisition device support 4 is connected with the camera sliding device 6, the camera sliding device 6 is connected with the camera support 5, and the camera support 5 is connected with the camera 2; the adapter port of the camera 2 is connected with the camera The adapter of the lens 3 is connected, the light inlet of the camera lens 3 is connected with the emission filter holder 7, and the emission filter 8 is embedded in the draw-in slot of the emission filter holder 7; One end, the other end of the optical fiber 9 points to the object to be observed.
图像处理部分中包括图像处理系统，与图像采集部分的相机2连接，接收白光图像及激发荧光图像，并对荧光图像自动分割、伪彩色添加、自动分割区域的光子数统计处理、自动分割区域的几何信息测量，将荧光图像同白光图像叠加后获得激发荧光成像区域的真实几何位置。相机1是能探测到微弱的荧光信号的光电探测器CCD相机。The image processing part includes an image processing system, connected with the camera 2 of the image acquisition part, receiving white light images and excited fluorescence images, and automatically segmenting the fluorescence images, adding false colors, statistical processing of photon numbers in automatically segmented areas, and automatically segmented areas. Geometric information measurement, after superimposing the fluorescence image with the white light image, the real geometric position of the excited fluorescence imaging area is obtained. Camera 1 is a photodetector CCD camera that can detect weak fluorescent signals.
具体的如图所示，图像处理系统包括：前处理模块21、分析模块22和存储模块23。Specifically, as shown in the figure, the image processing system includes: a pre-processing module 21 , an analysis module 22 and a storage module 23 .
前处理模块21与图像采集部分的相机2的数据输出端口连接，对接收到的荧光图像进行荧光强度均匀校正处理与自体荧光干扰去除处理，而对接收到的白光图像不做处理。The pre-processing module 21 is connected to the data output port of the camera 2 of the image acquisition part, and performs fluorescence intensity uniformity correction processing and autofluorescence interference removal processing on the received fluorescence images, but does not process the received white light images.
分析模块22与前处理模块21连接，对前处理模块21发送的荧光图像依次进行自动分割、伪彩色添加、自动分割区域的光子数统计、自动分割区域的几何信息测量、与白光图像叠加的操作，并显示荧光图像经过自动分割、伪彩色添加、自动分割区域的光子数统计并与白光图像叠加后得到的图像。The analysis module 22 is connected to the pre-processing module 21, and sequentially performs automatic segmentation, pseudo-color addition, photon number statistics of the automatically segmented area, geometric information measurement of the automatically segmented area, and superimposition with the white light image on the fluorescence image sent by the pre-processing module 21 , and display the image obtained after the fluorescence image is automatically segmented, pseudo-color added, photon number statistics of the automatically segmented area, and superimposed with the white light image.
存储模块23与前处理模块21和分析模块22连接，对前处理模块21处理后的荧光图像和白光图像进行保存，并对分析模块22处理后的，即在荧光图像经过自动分割、伪彩色添加、自动分割区域的光子数统计并与白光图像叠加后得到的图像进行保存。The storage module 23 is connected to the pre-processing module 21 and the analysis module 22, and stores the fluorescence image and the white light image processed by the pre-processing module 21, and processes the fluorescence image processed by the analysis module 22, that is, after the fluorescence image is automatically segmented and added with false color , The number of photons in the automatically segmented area is counted and the image obtained after superimposing it with the white light image is saved.
图2为本发明分子影像成像验证方法的流程图，如图所示，本方法具体包括如下步骤：Fig. 2 is a flowchart of the molecular imaging verification method of the present invention, as shown in the figure, the method specifically includes the following steps:
步骤S1：通过控制相机滑动装置及相机支架的位置，使相机镜头的测量区域准确对应被检测切片，相机镜头视野完整包括被检测切片横截面区域，实现相机对被检测切片的清晰成像；Step S1: By controlling the position of the camera sliding device and the camera bracket, the measurement area of the camera lens accurately corresponds to the detected slice, and the field of view of the camera lens completely includes the cross-sectional area of the detected slice, so that the camera can clearly image the detected slice;
具体的，就是软件及硬件初始化操作。将相机2温度锁定到-70℃，以降低图像噪声；Specifically, it is software and hardware initialization operations. Lock the temperature of camera 2 to -70°C to reduce image noise;
首先将待检测小动物注射荧光探针，然后进行冰冻，待冰冻完成后取出放到冰冻切片机内；Firstly, the small animal to be detected is injected with a fluorescent probe, then frozen, and taken out and placed in a cryostat after the freezing is completed;
通过调节相机支架5的角度以及相机滑动装置6，调节相机到正对切片的位置。调节相机镜头的光圈，实现切片的清晰成像。相机2与被检测切片间的距离调节为20cm。By adjusting the angle of the camera support 5 and the camera sliding device 6, adjust the camera to the position facing the slice. Adjust the aperture of the camera lens to achieve clear imaging of slices. The distance between the camera 2 and the detected slice was adjusted to 20cm.
步骤S2：开启冰冻切片机白光灯，控制相机2获取白光图像。白光图像反映被检测切片的横截面信息；Step S2: turn on the white light lamp of the cryostat, and control the camera 2 to acquire white light images. The white light image reflects the cross-sectional information of the detected slice;
步骤S3：将发射滤光片8嵌入发射滤光片支架7，并连接发射滤光片支架7和相机镜头3；开启激发光源10，同时关闭白光灯；控制相机2获取荧光图像；关闭激发光源10；荧光图像反应被检测切片包含的荧光光源的分布信息；Step S3: Insert the emission filter 8 into the emission filter holder 7, and connect the emission filter holder 7 and the camera lens 3; turn on the excitation light source 10, and turn off the white light at the same time; control the camera 2 to obtain a fluorescent image; turn off the excitation light source 10. The fluorescent image reflects the distribution information of the fluorescent light source contained in the detected slice;
步骤S4:荧光图像和白光图像传送到图像处理部分的前处理模块21；前处理模块21对荧光图像进行强度校正操作和自体荧光去除处理操作；Step S4: the fluorescence image and the white light image are sent to the pre-processing module 21 of the image processing part; the pre-processing module 21 performs an intensity correction operation and an autofluorescence removal processing operation on the fluorescence image;
步骤S5：分析模块22对前处理模块21发送的荧光图像依次进行自动分割、伪彩色添加、自动分割区域的光子数统计、自动分割区域的几何信息测量、与白光图像叠加等一系列操作，并显示荧光图像经过自动分割、伪彩色添加、自动分割区域的光子数统计并与白光图像叠加后得到的图像；Step S5: The analysis module 22 sequentially performs a series of operations on the fluorescent image sent by the pre-processing module 21, such as automatic segmentation, pseudo-color addition, photon number statistics of the automatically segmented area, geometric information measurement of the automatically segmented area, and superimposition with the white light image, and Display the image obtained after automatic segmentation, pseudo-color addition, photon number statistics of the automatically segmented area, and superimposition with the white light image of the fluorescence image;
步骤S6：存储模块23对前处理模块21处理后的荧光图像和白光图像进行保存，并对分析模块22处理后的，即在荧光图像经过自动分割、伪彩色添加、自动分割区域的光子数统计并与白光图像叠加后得到的图像进行保存，并保存测量得到的荧光区域几何信息。将相机2温度升高到25℃，硬件去初始化操作。Step S6: the storage module 23 saves the fluorescent image and the white light image processed by the pre-processing module 21, and counts the number of photons in the fluorescent image processed by the analysis module 22, that is, after automatic segmentation, pseudo-color addition, and automatic segmentation And save the image obtained after being superimposed with the white light image, and save the geometric information of the fluorescent area obtained from the measurement. Raise the temperature of camera 2 to 25°C, and de-initialize the hardware.
一次验证过程中，各步骤中人工控制极少，图像处理部分各模块所执行的所有操作及顺序均在软件中设置完毕。用户在连接系统完成后可不用关心整个验证过程的具体细节操作，便可以得到所需要的切片几何信息。此操作一体化设计大大方便了用户。In a verification process, there is very little manual control in each step, and all operations and sequences performed by each module in the image processing part are set in the software. After the connection system is completed, the user does not need to care about the specific details of the entire verification process, and can obtain the required slice geometry information. The integrated design of this operation greatly facilitates the user.
本发明的有益效果是：建立了一套结合冰冻切片机及CCD相机的分子影像成像验证系统及验证方法。该系统及方法能够完成对小动物等待检测物体的横截面切片测量，在设定参数后可自动获取清晰的小动物切片荧光数据，完成白光图像采集、荧光图像采集及与白光图像叠加、荧光图像自动分割、分割区域光子数统计、荧光区域几何信息测量等处理，大大简化了操作步骤和操作流程，为分子影像成像结果提供金标准。本发明系统结构合理，功能显著，操作方便，可广泛应用于光学分子成像领域，具有广阔的市场前景。The beneficial effect of the invention is that a set of molecular image imaging verification system and verification method combined with a cryostat and a CCD camera is established. The system and method can complete the measurement of cross-sectional slices of small animals waiting to be detected, and can automatically obtain clear fluorescence data of small animal slices after setting parameters, and complete white light image acquisition, fluorescence image acquisition and superimposition with white light images, and fluorescence image acquisition. Automatic segmentation, statistics of photon counts in segmented areas, and geometric information measurement of fluorescent areas greatly simplifies the operation steps and procedures, providing the gold standard for molecular imaging results. The system of the invention has reasonable structure, significant functions and convenient operation, can be widely used in the field of optical molecular imaging, and has broad market prospects.
以上所述的具体实施方式，对本发明的目的、技术方案和有益效果进行了进一步详细说明，所应理解的是，以上所述仅为本发明的具体实施方式而已，并不用于限定本发明的保护范围，凡在本发明的精神和原则之内，所做的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。The specific embodiments described above have further described the purpose, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above descriptions are only specific embodiments of the present invention and are not intended to limit the scope of the present invention. Protection scope, within the spirit and principles of the present invention, any modification, equivalent replacement, improvement, etc., shall be included in the protection scope of the present invention.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109799214A (en) * | 2018-12-14 | 2019-05-24 | 南京巨鲨显示科技有限公司 | A kind of optical test box |
| CN111413342A (en) * | 2020-04-20 | 2020-07-14 | 华中科技大学 | Microscopic imaging system for frozen section |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113516649B (en) * | 2021-07-28 | 2024-02-02 | 亿嘉和科技股份有限公司 | Cabinet surface detection method based on super-pixel segmentation |
| CN114136937A (en) * | 2021-11-16 | 2022-03-04 | 黑龙江省微甄光电科技有限责任公司 | Multifunctional miniature fluorescent microscopic imaging device and imaging method thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5422730A (en) * | 1994-03-25 | 1995-06-06 | Barlow; Clyde H. | Automated optical detection of tissue perfusion by microspheres |
| US20050182321A1 (en) * | 2002-03-12 | 2005-08-18 | Beth Israel Deaconess Medical Center | Medical imaging systems |
| US20080090198A1 (en) * | 2006-10-13 | 2008-04-17 | Rongguang Liang | Apparatus for caries detection |
| CN102106723A (en) * | 2011-03-17 | 2011-06-29 | 中国科学院自动化研究所 | Fluorescence molecule imaging device |
| CN102151122A (en) * | 2011-03-17 | 2011-08-17 | 中国科学院自动化研究所 | Laser fluorescent molecular imaging system and an instant fluorescent imaging method |
| CN202351599U (en) * | 2011-11-01 | 2012-07-25 | 浙江吉利汽车研究院有限公司 | Mounting bracket |
| CN103385696A (en) * | 2013-07-24 | 2013-11-13 | 中国科学院自动化研究所 | Fluorescence excitation real-time imaging system and method |
| CN103743714A (en) * | 2014-01-14 | 2014-04-23 | 苏州大猫单分子仪器研发有限公司 | Inclined wide-field optical section scanning imaging microscope system and imaging method thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101766476B (en) * | 2009-07-08 | 2011-05-11 | 中国科学院自动化研究所 | Auto-fluorescence molecule imaging system |
| CN102590155A (en) * | 2012-01-16 | 2012-07-18 | 华中科技大学 | Tissue slice scanning and imaging device |
| US9164015B2 (en) * | 2012-06-29 | 2015-10-20 | General Electric Company | Systems and methods for processing and imaging of biological samples |
-
2014
- 2014-09-19 CN CN201410482618.6A patent/CN104181142A/en active Pending
- 2014-09-22 WO PCT/CN2014/087072 patent/WO2016041211A1/en active Application Filing
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5422730A (en) * | 1994-03-25 | 1995-06-06 | Barlow; Clyde H. | Automated optical detection of tissue perfusion by microspheres |
| US20050182321A1 (en) * | 2002-03-12 | 2005-08-18 | Beth Israel Deaconess Medical Center | Medical imaging systems |
| US20080090198A1 (en) * | 2006-10-13 | 2008-04-17 | Rongguang Liang | Apparatus for caries detection |
| CN102106723A (en) * | 2011-03-17 | 2011-06-29 | 中国科学院自动化研究所 | Fluorescence molecule imaging device |
| CN102151122A (en) * | 2011-03-17 | 2011-08-17 | 中国科学院自动化研究所 | Laser fluorescent molecular imaging system and an instant fluorescent imaging method |
| CN202351599U (en) * | 2011-11-01 | 2012-07-25 | 浙江吉利汽车研究院有限公司 | Mounting bracket |
| CN103385696A (en) * | 2013-07-24 | 2013-11-13 | 中国科学院自动化研究所 | Fluorescence excitation real-time imaging system and method |
| CN103743714A (en) * | 2014-01-14 | 2014-04-23 | 苏州大猫单分子仪器研发有限公司 | Inclined wide-field optical section scanning imaging microscope system and imaging method thereof |
Non-Patent Citations (4)
| Title |
|---|
| 史毓阶: "《汉英医学装备科学仪器分类词典》", 31 March 2000, 中国医药科技出版社 * |
| 李原等: "《计算机辅助几何设计技术与应用》", 31 January 2007, 西北工业大学出版社 * |
| 王云双等: "《临床免疫学检验》", 31 March 2009, 军事医学科学出版社 * |
| 陈智毅等: "《分子影像学：基础与应用》", 31 January 2013, 广东高等教育出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109799214A (en) * | 2018-12-14 | 2019-05-24 | 南京巨鲨显示科技有限公司 | A kind of optical test box |
| CN111413342A (en) * | 2020-04-20 | 2020-07-14 | 华中科技大学 | Microscopic imaging system for frozen section |
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|---|---|
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