高分辨率X射线成像技术与应用研究
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摘要
随着高亮度的同步辐射光源的发展和纳米加工技术的飞速进步,基于波带片的高分辨率X射线成像技术在近十几年来发展极为迅速,其空间分辨率已达到15-50纳米。高分辨率同步辐射X射线成像技术具有穿透能力强、成像衬度来源丰富、无需复杂的制样和真空环境、三维成像能力等优势,弥补了其它显微术的不足(如光学显微镜和电镜),在生物医学、环境科学、材料科学和工业上有着广泛的应用前景。国家同步辐射实验室成功建造了一条高分辨率全场透射式X射线成像线站,空间分辨率达到50纳米。在此实验平台基础上,本论文主要开展了以下几个方面的工作：
1.高分辨率X射线成像线站的建设、测试及其成像原理研究
介绍了高分辨率X射线成像光束线和系统的光学设计。测试得到了光束线的能量分辨率、光子通量和实验站的成像空间分辨率等重要实际性能参数,测试结果表明高分辨率X射线成像线站的性能达到了设计指标。对高分辨率全场透射式X射线成像技术的工作原理进行了系统的研究。结合成像线站的设计,对系统的光路传播进行了分析,给出了光路传播方程的数学描述,简化了光路模拟过程。同时通过成像实验、理论分析和计算机模拟,分析成像系统的空间分辨与光学元件、照明光源、噪音和CCD探测器之间的关系。为将来发展新成像方法提供了理论基础。
2.高分辨率X射线Zernike相位衬度成像
衬度和分辨率是X射线成像系统的两个重要参数。提高系统的衬度,特别是对于生物样品或轻元素样品来说,具有非常重要的意义。因为轻元素折射率中的吸收项太小,X射线吸收衬度成像不能提供足够的衬度。而轻元素折射率中的相位项比吸收项大三到四个量级,利用相位衬度成像将能够提供比吸收衬度成像高得多的衬度。本工作在高分辨率X射线成像系统上实现了Zernike相位衬度成像实验方法,通过一系列样品分析了该方法的适用范围,并分析了halo现象。
3.高分辨率X射线三维成像技术和大视场三维成像技术研究
结合计算机辅助断层成像技术(computerized tomography,简称CT),发展了高分辨率X射线三维成像技术,具备了观测厚样品内部纳米三维结构的能力。然而在提高成像系统的分辨率同时也降低了视场。大多数样品在具备纳米结构特征的同时,其尺寸大多超过系统的视场(15微米)。这使得发展一种方法,既能得到样品的纳米三维结构信息,又能成像足够大的样品以得到有效的实验数据,颇为重要。结合系统的性能参数,通过电机扫描对样品等间隔角度进行大视场成像,利用迭代算法和标准滤波反投影算法进行三维重构,发展了大视场三维成像技术。实验结果表明该方法达到了预期要求。
4.定量计算方法
发展了对X射线三维数据进行定量计算的方法。根据Beer定律,X射线穿过物体后,物体对入射X射线的吸收效应是线性的。采集到得投影图的光强分布是物体吸收系数在X射线传播方向上积分(或投影)的信息。只要采集到完备的三维数据,重构后就可以定量地解出样品的吸收系数的三维分布。通过对断层数据进行取直方图、分析归类和统计计算等步骤,可以对样品的三维数据进行定量计算。
5.纳米材料的三维结构表征研究
设计、建模和表征始终是纳米材料科学设计流程中联系最紧密的三个环节。其中设计和建模最终指导制造,而表征(如成像)则是提供数据,证实设计和建模环节的可行性,或为优化设计提供依据。可见表征手段的重要性。本工作首次将高分辨率X射线三维成像技术成功应用到纳米材料科学中。通过对‘几何明星’凹陷Escher型硫化铜十四面体等纳米材料和燃料电池等纳米化器件进行了三维成像,得到了独特的样品三维信息。
6.酵母细胞的三维结构表征研究
新的成像方法,极大推进了我们对细胞的结构和功能的理解。每种成像方法都有其优缺点,没有一个成像方法能得到细胞的全部信息。本工作结合重金属染色的方法,在X射线能量为5.4 keV的条件下,成功得到了酵母细胞的三维结构。结合同步辐射高分辨率X射线成像技术和X射线荧光技术,成功得到了富硒酵母细胞中硒纳米球颗粒的形貌和三维分布。
7.小鼠骨小梁的纳米三维结构表征研究
骨头的三维结构和矿化度是其两个重要参数,与骨头的力学性能和骨质疏松症等疾病有着极为密切的关系。然而目前的大多数探测手段都局限于微米级的三维结构。本工作利用高分辨率X射线三维成像技术成功表征了小鼠骨小梁的纳米三维结构。得到了骨小梁中的腔隙和骨小管的三维结构和分布,并通过定量计算分析得到了骨小梁的矿化度。这为研究骨头的相关物理性能和骨质疏松症等疾病的病变原理提供了新的有力研究手段。
Due to the availability of synchrotron radiation source and the development of nano-fabrication technique, Fresnel zone-plates (FZPs) based X-ray microscopy has developed rapidly over the past decade, which offers spatial resolution in the 15-50 nm range. It complements other imaging techniques, such as optical and electron microscopy, by offering a unique set of capabilities including large penetration depth, high exposure efficiency, elemental and chemical specificity, magnetization sensitivity, as well as in-situ imaging with applied fields, overcoatings, and wet environments. The outstanding feature of this technique highlights its potential in a broad variety of fields, including life sciences, environmental science, materials science and industry. National Synchrotron Radiation Laboratory (NSRL) has successfully constructed a high resolution full-field, transmission x-ray microscope (TXM) beamline and endstation with a resolution of 50 nm. The main work and innovations are described as the following:
1. Construction and test of high resolution x-ray microscope beamline and system, and theory of image formation
This section introduces the optical design of the high-resolution x-ray microscope beamline and system. After the successful construction of beamline and system, we get the key parameters such as the energy resolution, photon flux and spatial resolution through a series of tests. The test results show that the performance of high-resolution x-ray microscope beamline and system achieves the design targets. The theory of image formation of the high-resolution x-ray imaging system is also presented in this section. The optical propagation equation is described here, which simplifys the computer simulation. Based on both theoretical and experimental studies, we study the relationship between spatial resolution and optics, illumination, noise, CCD detector. The results would provide fundamental principles for developing new imaging method.
2. High-resolution x-ray Zernike phase-contrast imaging
Contrast and resolution are two important parameters of the X-ray miscroscopy. It has an extremely important significance to improve the contrast, especially for biological samples or samples consisting of light elements. X-ray absorption-contrast imaging can not provide sufficient contrast, because the imaginary part of refractive index of light elements is too small. However, the real part of refractive index of light elements is three or four orders magnitude than the imaginary part. Thus phase contrast imaging would provide much higher contrast than absorption imaging for light elements. We study the Zernike phase-contrast imaging mechanism, which were also achieved on the high-resolution X-ray microscope. The halo effect was also investigated.
3. High-resolution x-ray tomography and large field of view (FOV) tomography
Combining computerized tomography (CT), we have developed high-resolution x-ray tomography, which enable us to observe the three-dimensional structure of sample at nanometer level. While increasing the the resolution of microscope, we sacrifice the field of view. However, the sizes of most samples with characteristics nano-structures beyond the field of view (15μm). Here we develop a large field of view (FOV) tomography, which can get the 3D structure of large enough volume with the same resolution. The reconstruction was performed using both the iterative algorithms and standard filtered-back-projection algorithm. The experimental results show that the method achieves the expected demand.
4. Quantitative analysis of tomographic data
According to Beer's Law, the x-ray absorption effect of the object is linear after it passes through objects. The intensity distribution of projection is the integration of absorption coefficient along the x-ray propagation direction. The three-dimensional distribution of absorption coefficient of the sample can be solved if a complete tomographic data is taken. Quantitative analysis can be performed on the tomographic data after procedures such as histogram, setting threshold and statistical calculation.
5. X-ray tomography of the nanomaterials
Design, modeling, and characterization technologies together are intimate components of the design cycle in technology development. Design and modeling are closely intertwined, ultimately guiding fabrication. Characterization technologies, imaging and measurement, provide the data that validate or drive revision of both designs and models. Characterization technologies are crucial. We get unique 3D information of nanomaterials and nano-enabled devices such as concaved cuboctahedron copper sulfide crystal and solid oxide fuel cell.
6. X-ray tomography of the yeast cells
New imaging methods have greatly advanced our understanding of cell structure and function. However, each of these imaging methods has its pros and cons. No single imaging method proves to be the perfect solution. Combining simple chemical treatment with absorption contrast imaging at 5.4 keV, the ultrastructural details of S. pombe were well delineated. Combining high resolution X-ray tomography with X-ray fluorescent probe, the shape and 3D distribution of elemental selenium nano particles were detected in selenium enriched yeast.
7. X-ray tomo graphy of the trabecula bone
Three-dimensional structure and mineralization of bone are two important parameters, which are closely related to its mechanical parameters and symptoms such as osteoporosis. However, most of the current detection methods are limited to micron-level 3D structure. Here high-resolution x-ray microscopy has been used to characterize the nano 3D structure of trabecular bone. The 3D distribution of lacuna and canaliculi system in trabecular bone has been gotten. The mineralization degree of trabecular bone has also been gotten through quantitative analysis. It would be a powerful tool for studying the physical properties of bone and symptoms such as osteoporosis.
1.高分辨率X射线成像线站的建设、测试及其成像原理研究
介绍了高分辨率X射线成像光束线和系统的光学设计。测试得到了光束线的能量分辨率、光子通量和实验站的成像空间分辨率等重要实际性能参数,测试结果表明高分辨率X射线成像线站的性能达到了设计指标。对高分辨率全场透射式X射线成像技术的工作原理进行了系统的研究。结合成像线站的设计,对系统的光路传播进行了分析,给出了光路传播方程的数学描述,简化了光路模拟过程。同时通过成像实验、理论分析和计算机模拟,分析成像系统的空间分辨与光学元件、照明光源、噪音和CCD探测器之间的关系。为将来发展新成像方法提供了理论基础。
2.高分辨率X射线Zernike相位衬度成像
衬度和分辨率是X射线成像系统的两个重要参数。提高系统的衬度,特别是对于生物样品或轻元素样品来说,具有非常重要的意义。因为轻元素折射率中的吸收项太小,X射线吸收衬度成像不能提供足够的衬度。而轻元素折射率中的相位项比吸收项大三到四个量级,利用相位衬度成像将能够提供比吸收衬度成像高得多的衬度。本工作在高分辨率X射线成像系统上实现了Zernike相位衬度成像实验方法,通过一系列样品分析了该方法的适用范围,并分析了halo现象。
3.高分辨率X射线三维成像技术和大视场三维成像技术研究
结合计算机辅助断层成像技术(computerized tomography,简称CT),发展了高分辨率X射线三维成像技术,具备了观测厚样品内部纳米三维结构的能力。然而在提高成像系统的分辨率同时也降低了视场。大多数样品在具备纳米结构特征的同时,其尺寸大多超过系统的视场(15微米)。这使得发展一种方法,既能得到样品的纳米三维结构信息,又能成像足够大的样品以得到有效的实验数据,颇为重要。结合系统的性能参数,通过电机扫描对样品等间隔角度进行大视场成像,利用迭代算法和标准滤波反投影算法进行三维重构,发展了大视场三维成像技术。实验结果表明该方法达到了预期要求。
4.定量计算方法
发展了对X射线三维数据进行定量计算的方法。根据Beer定律,X射线穿过物体后,物体对入射X射线的吸收效应是线性的。采集到得投影图的光强分布是物体吸收系数在X射线传播方向上积分(或投影)的信息。只要采集到完备的三维数据,重构后就可以定量地解出样品的吸收系数的三维分布。通过对断层数据进行取直方图、分析归类和统计计算等步骤,可以对样品的三维数据进行定量计算。
5.纳米材料的三维结构表征研究
设计、建模和表征始终是纳米材料科学设计流程中联系最紧密的三个环节。其中设计和建模最终指导制造,而表征(如成像)则是提供数据,证实设计和建模环节的可行性,或为优化设计提供依据。可见表征手段的重要性。本工作首次将高分辨率X射线三维成像技术成功应用到纳米材料科学中。通过对‘几何明星’凹陷Escher型硫化铜十四面体等纳米材料和燃料电池等纳米化器件进行了三维成像,得到了独特的样品三维信息。
6.酵母细胞的三维结构表征研究
新的成像方法,极大推进了我们对细胞的结构和功能的理解。每种成像方法都有其优缺点,没有一个成像方法能得到细胞的全部信息。本工作结合重金属染色的方法,在X射线能量为5.4 keV的条件下,成功得到了酵母细胞的三维结构。结合同步辐射高分辨率X射线成像技术和X射线荧光技术,成功得到了富硒酵母细胞中硒纳米球颗粒的形貌和三维分布。
7.小鼠骨小梁的纳米三维结构表征研究
骨头的三维结构和矿化度是其两个重要参数,与骨头的力学性能和骨质疏松症等疾病有着极为密切的关系。然而目前的大多数探测手段都局限于微米级的三维结构。本工作利用高分辨率X射线三维成像技术成功表征了小鼠骨小梁的纳米三维结构。得到了骨小梁中的腔隙和骨小管的三维结构和分布,并通过定量计算分析得到了骨小梁的矿化度。这为研究骨头的相关物理性能和骨质疏松症等疾病的病变原理提供了新的有力研究手段。
Due to the availability of synchrotron radiation source and the development of nano-fabrication technique, Fresnel zone-plates (FZPs) based X-ray microscopy has developed rapidly over the past decade, which offers spatial resolution in the 15-50 nm range. It complements other imaging techniques, such as optical and electron microscopy, by offering a unique set of capabilities including large penetration depth, high exposure efficiency, elemental and chemical specificity, magnetization sensitivity, as well as in-situ imaging with applied fields, overcoatings, and wet environments. The outstanding feature of this technique highlights its potential in a broad variety of fields, including life sciences, environmental science, materials science and industry. National Synchrotron Radiation Laboratory (NSRL) has successfully constructed a high resolution full-field, transmission x-ray microscope (TXM) beamline and endstation with a resolution of 50 nm. The main work and innovations are described as the following:
1. Construction and test of high resolution x-ray microscope beamline and system, and theory of image formation
This section introduces the optical design of the high-resolution x-ray microscope beamline and system. After the successful construction of beamline and system, we get the key parameters such as the energy resolution, photon flux and spatial resolution through a series of tests. The test results show that the performance of high-resolution x-ray microscope beamline and system achieves the design targets. The theory of image formation of the high-resolution x-ray imaging system is also presented in this section. The optical propagation equation is described here, which simplifys the computer simulation. Based on both theoretical and experimental studies, we study the relationship between spatial resolution and optics, illumination, noise, CCD detector. The results would provide fundamental principles for developing new imaging method.
2. High-resolution x-ray Zernike phase-contrast imaging
Contrast and resolution are two important parameters of the X-ray miscroscopy. It has an extremely important significance to improve the contrast, especially for biological samples or samples consisting of light elements. X-ray absorption-contrast imaging can not provide sufficient contrast, because the imaginary part of refractive index of light elements is too small. However, the real part of refractive index of light elements is three or four orders magnitude than the imaginary part. Thus phase contrast imaging would provide much higher contrast than absorption imaging for light elements. We study the Zernike phase-contrast imaging mechanism, which were also achieved on the high-resolution X-ray microscope. The halo effect was also investigated.
3. High-resolution x-ray tomography and large field of view (FOV) tomography
Combining computerized tomography (CT), we have developed high-resolution x-ray tomography, which enable us to observe the three-dimensional structure of sample at nanometer level. While increasing the the resolution of microscope, we sacrifice the field of view. However, the sizes of most samples with characteristics nano-structures beyond the field of view (15μm). Here we develop a large field of view (FOV) tomography, which can get the 3D structure of large enough volume with the same resolution. The reconstruction was performed using both the iterative algorithms and standard filtered-back-projection algorithm. The experimental results show that the method achieves the expected demand.
4. Quantitative analysis of tomographic data
According to Beer's Law, the x-ray absorption effect of the object is linear after it passes through objects. The intensity distribution of projection is the integration of absorption coefficient along the x-ray propagation direction. The three-dimensional distribution of absorption coefficient of the sample can be solved if a complete tomographic data is taken. Quantitative analysis can be performed on the tomographic data after procedures such as histogram, setting threshold and statistical calculation.
5. X-ray tomography of the nanomaterials
Design, modeling, and characterization technologies together are intimate components of the design cycle in technology development. Design and modeling are closely intertwined, ultimately guiding fabrication. Characterization technologies, imaging and measurement, provide the data that validate or drive revision of both designs and models. Characterization technologies are crucial. We get unique 3D information of nanomaterials and nano-enabled devices such as concaved cuboctahedron copper sulfide crystal and solid oxide fuel cell.
6. X-ray tomography of the yeast cells
New imaging methods have greatly advanced our understanding of cell structure and function. However, each of these imaging methods has its pros and cons. No single imaging method proves to be the perfect solution. Combining simple chemical treatment with absorption contrast imaging at 5.4 keV, the ultrastructural details of S. pombe were well delineated. Combining high resolution X-ray tomography with X-ray fluorescent probe, the shape and 3D distribution of elemental selenium nano particles were detected in selenium enriched yeast.
7. X-ray tomo graphy of the trabecula bone
Three-dimensional structure and mineralization of bone are two important parameters, which are closely related to its mechanical parameters and symptoms such as osteoporosis. However, most of the current detection methods are limited to micron-level 3D structure. Here high-resolution x-ray microscopy has been used to characterize the nano 3D structure of trabecular bone. The 3D distribution of lacuna and canaliculi system in trabecular bone has been gotten. The mineralization degree of trabecular bone has also been gotten through quantitative analysis. It would be a powerful tool for studying the physical properties of bone and symptoms such as osteoporosis.
引文
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