分子超光谱成像的生物组织定量检测与方法研究
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摘要
超光谱成像技术同时提供生物组织样本图谱两方面的信息,对检测目标可进行定性、定量和定位的描述,实现对某些病理变化的早期诊断,相比较于其他医学成像技术,具有独特的优势。本文结合超光谱成像技术在生物医学上的应用,设计了分子超光谱成像系统。由于该系统可以对样本进行无损伤、无接触检测,因而可用于生物切片或活体组织的定量检测和评估。本论文中,对自行研制的分子超光谱成像系统的成像机理和生物医学应用进行了深入综合地研究,取得了如下几点具有创新的成果：
1.在第一代推扫式显微高光谱成像光谱仪的基础上,改进了系统的分光方式,由推扫式成像改为基于AOTF(acousto-optic tunable filters)分光方式的成像。相比较与第一代成像系统,新一代成像系统具有快速电调谐、无活动部件、透光率比较高,成像质量好和可实时成像等特点。对系统的关键部件进行了研究,分析了系统的整体技术指标和性能,为分子超光谱数据的预处理和定量化提供依据。
2.结合系统各部件的性能和技术指标,开发系统软件,使软件能有效配合系统硬件采集超光谱图像数据。采集方式有多种,包括固定波段非积分方式连续采集、积分方式单帧采集、积分方式可变波段连续采集和非积分方式可变波段连续采集等,可根据实际应用进行灵活的选用。系统软件界面友好,参数配置灵活。
3.在分析分子超光谱成像系统成像的光学模型和数据模型的基础上,结合分子超光谱成像系统的性能,对生物组织的分子超光谱图像数据进行预处理。提出利用参考空白图像对生物组织的超光谱图像进行空间维和光谱维联合辐射校正,以消除了系统光源的影响,在ENVI和IDL软件平台上实现该算法；对数据进行归一化处理,使该系统测量的生物组织样本数据具有可比性。
4.对分子超光谱成像系统应用于坐骨神经中运动神经和感觉神经的分类进行了探索。在进行神经损伤修复时,只有将远端和近端相匹配的神经纤维对应地进行镜面吻合,才能保证神经修复的效果。使用分子超光谱成像系统多次采集神经切片样本,分别从神经纤维的外膜、整个神经纤维、神经纤维的内核、神经纤维的外环四个不同的类型区域寻找特征光谱,分析其光谱特性及分类结果,探索用超光谱成像技术应用于运动神经和感觉神经分类的可行性。
5.在第一代推扫式显微高光谱成像仪对糖尿病视网膜病的研究进行探索的基础上,将分子超光谱成像系统应用于EPO(促红细胞生长素)和AAV2-EPO(腺相关病毒2型介导的促红细胞生长素)对实验室大鼠早期糖尿病视网膜病变的疗效研究中。采集了正常对照组N、糖尿病组D、糖尿病EPO治疗组E0、糖尿病AAV2-EPO治疗组(高剂量)E1、糖尿病AAV2-EPO治疗组(中剂量)E2、糖尿病AAV2-EPO治疗组(低剂量)E3的大鼠视网膜组织切片的分子超光谱图像,使用建立的模型对获得的数据进行了预处理,不仅从图像和光谱的角度定性分析了EPO和AAV2-EPO对大鼠糖网病具有一定的疗效,还从外核层厚度计算、光谱的相对误差比较、光谱相似性比较三个方面定量分析EPO和AAV2-EPO对大鼠糖网病的治疗效果。
The hyperspectral imaging technology can provide information of image and spectra about biological tissue samples simultaneously, and give qualitative, quantitative and positioning description aim at the detection target, and achieve early diagnosis of certain pathological changes. Compared to other medical imaging technology, it has unique advantages. In this paper, a molecular hyperspectral imaging (MHSI) system was designed with hyperspectral imaging technology. Since the system can detect sample without damage and contact detection, it has been used for biological detection and quantitative assessment of living tissue. By depth comprehensive study the self-developed MHSI system about molecular hyperspectral imaging principle and biomedical imaging applications, the following innovative results were acquired:
1. On the basis of the first generation of microscopic pushbroom hyperspectral imaging system, a novel MHSI system based on AOTF was presented. Compared with the first generation of imaging systems, the second generation has superior performance with fast electrical tuning, no moving parts, relatively high transmission rate, good image quality and real-time imaging. By studying the key components, we analyzed the system's overall specifications and performance in order to provide the basis for preprocessing and quantification of molecular hyperspectral imaging data.
2. Combination of the system performance and technical indicators of various components, we developed system software. The system software can effectively run with the system hardware and capture the molecular hyperspectral imaging data by different ways, including fixed-band continuous acquisition of non-integral approach, single-frame capture mode of integral approach, continuous variable-band approach collection of integral approach and continuous variable-band collection of non-integration approach, etc. These ways can be flexibly chosen. System software has friendly interface to flexibly configurate parameters.
3. In the analysis of optical model and data model of MHSI system, the molecular hyperspectral imaging data of biological issue are preprocessed referring performance of MHSI system. A new gray correction coefficient algorithm with spatial dimension and spectral dimension was provided by using reference blank images of biological tissue. The algorithm can effectively eliminate the influence of system light. It was achieved in ENVI and IDL software platform. The data was normalized so that it was comparable with the biological tissue sample data.
4. We are taking vigorous action to explore a new way of classify motor and sensory nerve by MPHI system. During nerve repair, only the distal and proximal nerve fibers are matched with corresponding to the mirror, in order to ensure the effect of nerve repair. We captured the nerve biopsy samples by MHSI system, and found four different types of the spectral characteristics of motor and sensory nerve, respectively from the outer membrane of nerve fibers, the nerve fibers, the core of nerve fibers, the outer ring of nerve fiber. We also analysed their spectral characteristics and classification and explored feasibility to classification of motor and sensory nerve by MHSI system.
5. Base on the research of diabetic retinopathy(DR) by the microscopic pushbroom hyperspectral imaging system(MPHI), the relative work wa still go on to evaluate the protective effects of erythropoietin(EPO) and Adeno-Associated Virus Type 2 erythropoietin (AAV2-EPO) on early diabetic retinopathy in rats byMHSI system. Molecular hyperspectral imaging data are collected and preprocessed from six groups:normal(N),diabetic(D),EPO-treated(E0),AAV2-EPO(low-dose)-treated(E1), AAV2-EPO(middle-dose)-treated(E2) and AAV2-EPO(high-dose)-treated(E3). Not only by qualitative analysis of images and spectra curves, but also by quantitative analysis of the thickness of ONL, the comparision of relative errors and the comparision of the spectral similarity, the results suggest EPO and AAV2-EPO may prevent cell apoptosis and has protective effect on rat DR at a certain level.
1.在第一代推扫式显微高光谱成像光谱仪的基础上,改进了系统的分光方式,由推扫式成像改为基于AOTF(acousto-optic tunable filters)分光方式的成像。相比较与第一代成像系统,新一代成像系统具有快速电调谐、无活动部件、透光率比较高,成像质量好和可实时成像等特点。对系统的关键部件进行了研究,分析了系统的整体技术指标和性能,为分子超光谱数据的预处理和定量化提供依据。
2.结合系统各部件的性能和技术指标,开发系统软件,使软件能有效配合系统硬件采集超光谱图像数据。采集方式有多种,包括固定波段非积分方式连续采集、积分方式单帧采集、积分方式可变波段连续采集和非积分方式可变波段连续采集等,可根据实际应用进行灵活的选用。系统软件界面友好,参数配置灵活。
3.在分析分子超光谱成像系统成像的光学模型和数据模型的基础上,结合分子超光谱成像系统的性能,对生物组织的分子超光谱图像数据进行预处理。提出利用参考空白图像对生物组织的超光谱图像进行空间维和光谱维联合辐射校正,以消除了系统光源的影响,在ENVI和IDL软件平台上实现该算法；对数据进行归一化处理,使该系统测量的生物组织样本数据具有可比性。
4.对分子超光谱成像系统应用于坐骨神经中运动神经和感觉神经的分类进行了探索。在进行神经损伤修复时,只有将远端和近端相匹配的神经纤维对应地进行镜面吻合,才能保证神经修复的效果。使用分子超光谱成像系统多次采集神经切片样本,分别从神经纤维的外膜、整个神经纤维、神经纤维的内核、神经纤维的外环四个不同的类型区域寻找特征光谱,分析其光谱特性及分类结果,探索用超光谱成像技术应用于运动神经和感觉神经分类的可行性。
5.在第一代推扫式显微高光谱成像仪对糖尿病视网膜病的研究进行探索的基础上,将分子超光谱成像系统应用于EPO(促红细胞生长素)和AAV2-EPO(腺相关病毒2型介导的促红细胞生长素)对实验室大鼠早期糖尿病视网膜病变的疗效研究中。采集了正常对照组N、糖尿病组D、糖尿病EPO治疗组E0、糖尿病AAV2-EPO治疗组(高剂量)E1、糖尿病AAV2-EPO治疗组(中剂量)E2、糖尿病AAV2-EPO治疗组(低剂量)E3的大鼠视网膜组织切片的分子超光谱图像,使用建立的模型对获得的数据进行了预处理,不仅从图像和光谱的角度定性分析了EPO和AAV2-EPO对大鼠糖网病具有一定的疗效,还从外核层厚度计算、光谱的相对误差比较、光谱相似性比较三个方面定量分析EPO和AAV2-EPO对大鼠糖网病的治疗效果。
The hyperspectral imaging technology can provide information of image and spectra about biological tissue samples simultaneously, and give qualitative, quantitative and positioning description aim at the detection target, and achieve early diagnosis of certain pathological changes. Compared to other medical imaging technology, it has unique advantages. In this paper, a molecular hyperspectral imaging (MHSI) system was designed with hyperspectral imaging technology. Since the system can detect sample without damage and contact detection, it has been used for biological detection and quantitative assessment of living tissue. By depth comprehensive study the self-developed MHSI system about molecular hyperspectral imaging principle and biomedical imaging applications, the following innovative results were acquired:
1. On the basis of the first generation of microscopic pushbroom hyperspectral imaging system, a novel MHSI system based on AOTF was presented. Compared with the first generation of imaging systems, the second generation has superior performance with fast electrical tuning, no moving parts, relatively high transmission rate, good image quality and real-time imaging. By studying the key components, we analyzed the system's overall specifications and performance in order to provide the basis for preprocessing and quantification of molecular hyperspectral imaging data.
2. Combination of the system performance and technical indicators of various components, we developed system software. The system software can effectively run with the system hardware and capture the molecular hyperspectral imaging data by different ways, including fixed-band continuous acquisition of non-integral approach, single-frame capture mode of integral approach, continuous variable-band approach collection of integral approach and continuous variable-band collection of non-integration approach, etc. These ways can be flexibly chosen. System software has friendly interface to flexibly configurate parameters.
3. In the analysis of optical model and data model of MHSI system, the molecular hyperspectral imaging data of biological issue are preprocessed referring performance of MHSI system. A new gray correction coefficient algorithm with spatial dimension and spectral dimension was provided by using reference blank images of biological tissue. The algorithm can effectively eliminate the influence of system light. It was achieved in ENVI and IDL software platform. The data was normalized so that it was comparable with the biological tissue sample data.
4. We are taking vigorous action to explore a new way of classify motor and sensory nerve by MPHI system. During nerve repair, only the distal and proximal nerve fibers are matched with corresponding to the mirror, in order to ensure the effect of nerve repair. We captured the nerve biopsy samples by MHSI system, and found four different types of the spectral characteristics of motor and sensory nerve, respectively from the outer membrane of nerve fibers, the nerve fibers, the core of nerve fibers, the outer ring of nerve fiber. We also analysed their spectral characteristics and classification and explored feasibility to classification of motor and sensory nerve by MHSI system.
5. Base on the research of diabetic retinopathy(DR) by the microscopic pushbroom hyperspectral imaging system(MPHI), the relative work wa still go on to evaluate the protective effects of erythropoietin(EPO) and Adeno-Associated Virus Type 2 erythropoietin (AAV2-EPO) on early diabetic retinopathy in rats byMHSI system. Molecular hyperspectral imaging data are collected and preprocessed from six groups:normal(N),diabetic(D),EPO-treated(E0),AAV2-EPO(low-dose)-treated(E1), AAV2-EPO(middle-dose)-treated(E2) and AAV2-EPO(high-dose)-treated(E3). Not only by qualitative analysis of images and spectra curves, but also by quantitative analysis of the thickness of ONL, the comparision of relative errors and the comparision of the spectral similarity, the results suggest EPO and AAV2-EPO may prevent cell apoptosis and has protective effect on rat DR at a certain level.
引文
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