生物体内部结构三维显微成像技术研究
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摘要
生物体内部结构三维显微成像技术(Three-Dimensional Reconstruction of
Microscopic Images of Biological Internal Structure，简称3DRMI)的基本思想是将生物体进行序列化切片，得到二维序列图像，然后利用计算机对这些二维序列图像进行三维重构和显示，使生物体内部结构三维图像以及各断层面图像得到真实的再现。3DRMI技术涉及光学、机械、电子和计算机等多种技术，是多学科交叉融合的成果。该技术可广泛应用于生物学、生物医学及其相关领域。利用3DRMI技术，可以细致、完整、系统地获得生物体内部结构的数据信息，为后续研究提供依据。由于该技术的实用性及广阔应用前景，国内外的一些相关机构在进行研究，但从总体上来看，此项技术仍处于起步阶段。本文详细论述了3DRMI技术的基本原理和实现方法，介绍了已构建完成的3DRMI系统，从系统的角度对3DRMI技术进行深入的分析，其中着重研究了序列图像三维重构的算法及其实现，并给出了3DRMI实验研究及其结果。
本论文对硕士阶段的研究工作进行了总结，全文共分七章。第一章绪论，简要回顾几种内部结构成像技术的发展历史，并分析其发展现状及趋势；介绍本文的主要研究目标和研究内容。第二章3DRMI技术原理和方法，概述3DRMI成像系统的基本思想和总体框架；阐述了3DRMI技术涉及的切片技术、采集技术、三维重构技术的基本概念和基础知识。第三章序列图像的采集技术研究，介绍了显微成像系统、光照系统；设计了图像采集系统及其软硬件。第四章样品切片制备技术研究，内容包括序列切片系统机械装置设计，步进电机驱动的软硬件以及生物体样品制备技术研究。第五章序列图像的三维重构，首先对序列图像三维重构方法进行概述；其次，对二维图像处理软件进行研制；再次，对序列图像的三维几何重构的算法、程序模块以及序列图像三维重构程序进行研究和设计；最后介绍序列图像三维重构其他方法及其发展趋势。第六章生物体内部结构三维显微成像技术实验研究，是整个系统的实验部分，介绍了3DRMI实验系统；给出了模拟序列切片图像以及实际的简单序列切片图像的三维重构结果。第七章结论与展望，讨论了目前己构建的3DRMI系统存在的不足并提出了相应的改进设想。
Three-dimensional reconstruction of microscopic images (3DRMI) of internal structure of biological specimens is one of the most important research fields in biological engineering. It has achieved great successes in the world over last decade. However,in our country,the research of this technology is on the primary state. The technology of 3DRMI involves optics,mechanics,electronics and computer science. The procedure of 3DRMI includes the steps as follows:firstly,make section preparation of the biological sample;secondly,capture the serial images of the section plan and save them in sequence;thirdly,process the serial images;finally,reconstruct three-dimensional model of the serial images. Using 3DRMI system,we can get the data and three-dimensional inner or outer structure of any organ. Therefore,the research of 3DRMI is of great value. This thesis discusses the basic principle and the method of 3DRMI technology,introduces the 3DRMI system that we have built,and focuses on the research of the three-dimensi
onal reconstruction method of serial slice images. In the final part,we carry out the experiment on the 3DRMI system and present the result.
The thesis is organized as follows:
Chapter 1 introduces the history and development of several internal imaging methods. This chapter ends with the depiction of the primary research objectives of the thesis.
In Chapter 2,we discuss the essential principle and feature-based design of 3DRMI imaging system. The basic concepts and methods of the slice,capture and reconstruction technology closing to the 3DRMI are expounded.
Collection-technology on capturing a serial of slice image is introduced in Chapter 3. This part includes microscopic imaging system,lighting system,the hardware and software of capture system.
The prepare-technique of the serial sample slice is discussed in Chapter 4,
including the design of the slice machine,the software and hardware of the step-motor driving system. The technique of preparing the biological sample is discussed in this chapter.
Chapter 5 presents the three-dimensional reconstruction method of a serial slice images. This chapter is organized as following:firstly,introduce several 3D reconstruction methods;secondly,design the software of two-dimensional image processing;thirdly,analyze the algorithms of the three-dimensional reconstruction in detail and discuss the design and implementation of our serial image-reconstruction software system;finally,present the other reconstruction method and its development.
Chapter 6 introduces the 3DRMI experiment system and provides the corresponding experiment results. In this chapter,we simulate the 3D reconstruction of three serial images and give the reconstruction model of three serial actual slices images.
The final chapter summarizes the main results and makes conclusions of the thesis. Some future research work is also described.
Microscopic Images of Biological Internal Structure，简称3DRMI)的基本思想是将生物体进行序列化切片，得到二维序列图像，然后利用计算机对这些二维序列图像进行三维重构和显示，使生物体内部结构三维图像以及各断层面图像得到真实的再现。3DRMI技术涉及光学、机械、电子和计算机等多种技术，是多学科交叉融合的成果。该技术可广泛应用于生物学、生物医学及其相关领域。利用3DRMI技术，可以细致、完整、系统地获得生物体内部结构的数据信息，为后续研究提供依据。由于该技术的实用性及广阔应用前景，国内外的一些相关机构在进行研究，但从总体上来看，此项技术仍处于起步阶段。本文详细论述了3DRMI技术的基本原理和实现方法，介绍了已构建完成的3DRMI系统，从系统的角度对3DRMI技术进行深入的分析，其中着重研究了序列图像三维重构的算法及其实现，并给出了3DRMI实验研究及其结果。
本论文对硕士阶段的研究工作进行了总结，全文共分七章。第一章绪论，简要回顾几种内部结构成像技术的发展历史，并分析其发展现状及趋势；介绍本文的主要研究目标和研究内容。第二章3DRMI技术原理和方法，概述3DRMI成像系统的基本思想和总体框架；阐述了3DRMI技术涉及的切片技术、采集技术、三维重构技术的基本概念和基础知识。第三章序列图像的采集技术研究，介绍了显微成像系统、光照系统；设计了图像采集系统及其软硬件。第四章样品切片制备技术研究，内容包括序列切片系统机械装置设计，步进电机驱动的软硬件以及生物体样品制备技术研究。第五章序列图像的三维重构，首先对序列图像三维重构方法进行概述；其次，对二维图像处理软件进行研制；再次，对序列图像的三维几何重构的算法、程序模块以及序列图像三维重构程序进行研究和设计；最后介绍序列图像三维重构其他方法及其发展趋势。第六章生物体内部结构三维显微成像技术实验研究，是整个系统的实验部分，介绍了3DRMI实验系统；给出了模拟序列切片图像以及实际的简单序列切片图像的三维重构结果。第七章结论与展望，讨论了目前己构建的3DRMI系统存在的不足并提出了相应的改进设想。
Three-dimensional reconstruction of microscopic images (3DRMI) of internal structure of biological specimens is one of the most important research fields in biological engineering. It has achieved great successes in the world over last decade. However,in our country,the research of this technology is on the primary state. The technology of 3DRMI involves optics,mechanics,electronics and computer science. The procedure of 3DRMI includes the steps as follows:firstly,make section preparation of the biological sample;secondly,capture the serial images of the section plan and save them in sequence;thirdly,process the serial images;finally,reconstruct three-dimensional model of the serial images. Using 3DRMI system,we can get the data and three-dimensional inner or outer structure of any organ. Therefore,the research of 3DRMI is of great value. This thesis discusses the basic principle and the method of 3DRMI technology,introduces the 3DRMI system that we have built,and focuses on the research of the three-dimensi
onal reconstruction method of serial slice images. In the final part,we carry out the experiment on the 3DRMI system and present the result.
The thesis is organized as follows:
Chapter 1 introduces the history and development of several internal imaging methods. This chapter ends with the depiction of the primary research objectives of the thesis.
In Chapter 2,we discuss the essential principle and feature-based design of 3DRMI imaging system. The basic concepts and methods of the slice,capture and reconstruction technology closing to the 3DRMI are expounded.
Collection-technology on capturing a serial of slice image is introduced in Chapter 3. This part includes microscopic imaging system,lighting system,the hardware and software of capture system.
The prepare-technique of the serial sample slice is discussed in Chapter 4,
including the design of the slice machine,the software and hardware of the step-motor driving system. The technique of preparing the biological sample is discussed in this chapter.
Chapter 5 presents the three-dimensional reconstruction method of a serial slice images. This chapter is organized as following:firstly,introduce several 3D reconstruction methods;secondly,design the software of two-dimensional image processing;thirdly,analyze the algorithms of the three-dimensional reconstruction in detail and discuss the design and implementation of our serial image-reconstruction software system;finally,present the other reconstruction method and its development.
Chapter 6 introduces the 3DRMI experiment system and provides the corresponding experiment results. In this chapter,we simulate the 3D reconstruction of three serial images and give the reconstruction model of three serial actual slices images.
The final chapter summarizes the main results and makes conclusions of the thesis. Some future research work is also described.
引文
[1] 苟量，王绪本，曹辉，X射线成像技术的发展现状和趋势，成都理工学院学报，Vol．29(2)，2002；
[2] 贺国珠，李显耀，毕会英，核磁共振技术应用的基本原理，物理通报，Vol．12(7)，1997；
[3] 巫北海，CT与MRI进展，第三军医大学学报，Vol．24(6)，2002：627-630；
[4] H.D William, The World Medical Equipment, Vol.12(10), 1999;
[5] Brink JA. Technical aspects of helical (spiral) CT [J], Radiol Clin North Am, Vol.33(4), 1995;
[6] 沈彬 李加 张彦军 李宁，新型光学成像技术及其在生物医学中的应用，激光杂志，Vol．20(2)，1999；
[7] 特集，最近的显微镜，精密工学会杂志，Vol．57(7)，1991；
[8] Sjostrand FS, J Ultrastrust, Res, Vol.2, 1958;
[9] 邓利群，快速制作常规石蜡切片，湖北民族学院学报(医学版)，Vol．18(4)，2001；
[10] 曹敏年，一种用PLC控制步进电机的方法，上海电器技术，Vol4(17)，1996；
[11] 范正翘 刘进 程胜，单片机控制的步进电动机综合微步驱动系统，微特电机，Vol3．，1996；
[12] 郁道银 谈恒英，光学工程，机械工业出版社，5月，1999；
[13] 李晓彤，几何光学与光学设计，浙江大学出版社，7月，1997；
[14] Noriniko Nakano, et al., Digital Still Camera System for Megapixel CCD, IEEE Transaction on Consumer Electronics, 44(3), 1998;
[15] 陆勤编译，[美]Popular Photograghy，数字相机地实拍评价，照相机，Vol6；1998；
[16] 石教英 蔡文立著，科学计算可视化算法与系统，科学出版社，9，1996；
[17] Cline, H., Lorensen, W., et al., Two Algorithms for the 3D Reconstruction of Tomograms, Medical Physics, Vol. 15(3), 1988;
[18] Farin, G.E,Curves and Surface From Computer Aided Geometric Design, Academic Press, 1990;
[19] Levoy, M., Display of surfaces from Volume Data, IEEE CG&A, Vol.8(5), 1990;
[20] Fruhauf, M., Combining Volume Rendering with Line and Surface Rendering, Eurogriphics'91, 1991;
[21] Shinagawa, Y., Kunii, T.L., Constructing a Reeb Graph Automatically from Cross Section, IEEE CG&A, Vol. 11,1991;
[22] Christiansen, H.N., Sederberg, T.W., Construction of Complex contours Line Definition into Polygonal Element Mosaics, Computer Graphics, Vo1.12(3), 1978;
[23] 沈兰荪，图像编码与异步传输。北京：人民邮电出版社，1998；
[24] 林靖 陈辉堂 蒋平 王月娟，用多媒体视频卡实现实时数字图像采集，测控技术，Vol．18(2)，1999；
[25] 王永刚，卫保国，基于数码相机的图像采集系统，测控技术，Vol．19 No．5，2000
[26] Norihiko Nakano, et al., Digital Still Camera System for Megapixel CCD. IEEE transaction on Consumer Electronics, Vol.44(3), 1998;
[27] Mendis S K, et al. Progress in CMOS active pixel image sensors. SPIE Proceedings, 2172,1995;
[28] 陆勤编译，[美]Popular Photography，数字相机的实拍评价，照相机，Vol．6，1998；
[29] 谭玉柱 侯晓轩，三相反应式步进电机驱动接口电路设计及应用，中小型电机，27(1)2000；
[30] 陈瑛等，在微机上实现脑结构三维重构技术，中国医学影像技术，1999，15(12)；
[31] 王今著，生物组织显微镜结构三维重建的灰度阴影立体图对显示技术，生物物理学报，2(3)17，1990；
[32] Heffeman PB, Robb RA. A new method of shaded surface display of biological and medical image. IEEE Trans on Medical Imaging, MI-4 25, 1985;
[33] Udapa JK. Interactive segmentation and boundary surface formation for 3D digital images. Comput Graph Imag Proc, 18(15)213, 1982;
[34] 李淑梁，陈洁．三维CT图像的显示与重建．清华大学学报，26(5)10，1986；
[35] 张义，李时光，杨恬．生物组织显微切片图像计算机三维重建的截面重建技术，中国生物医学工程学报，11(1)17，1992；
[36] 陈qing，多目标离散断层图像序列的三维重建算法研究，重庆大学硕士学位论文，1995；
[37] 吕凤军，数字图像处理编程入门，清华大学出版社，1999．9；
[38] 束越新．《颜色光学基础理论》．济南：山东科学技术出版社，47-64，1981．
[39] 刘维一等，用空间图像融合技术进行三维重建的研究，光学学报，20(3)，2000；
[40] Christiansen, H.N., Sederberg, T.W., Construction of Complex contours Line Definition into Polygonal Element Mosaics, Computer Graphics, 12(3), 1978;
[41] Fchis, H., Kendem, Z., et al.,Optimal Surface Reconstruction from planar Contours, CACM, Vol. 20, 1979;
[42] Christiansen, H.N., Sederberg, T.W., Construction of Complex Contours Line Definition into Polygonal Element Mosaics, Computer Graphics, Vol. 12(3), 1978;
[43] Cook, L.T., Dwyer, S., et al., A Three Dimensional Display System for Diagnostic Imaging Applications, IEEE CG&A, Vol. 3(4), 1983;
[44] Gananpathy, S., Dennely, T.G., A new General Triangulation Method for Planar Contours, Computer Graphics, Vol. 16(3), 1982;
[45] Ekoule, A.B., Peyrin, F.C., et al., A Triangulation Algorithm from Arbitrary Shaped Multiple Planar Contours, ACM Trans. On Graphics, Vol. 10, 1991;
[46] Jaffery, S., Dutta, K., Digital Reconstruction Methods for 3D Image Visualization, SPIE, Vol.507, 1984;
[47] Sabella, P., A rendering Algorithm for Visualization 3D Scalar Fields, Computer Graphics, Vol. 22(4), 1988;
[48] Drebin, R.L., Carpenter R., et al., Volume Rendering, Computer Graphics, Vol. 18(3), 1984;
[49] Levoy, M., Display of Surfaces from Volume Data, IEEE CG&A, Vol. 8(5), 1988;
[50] Levoy, M., A Hybrid Ray Trace For Rendering Polygons and Volume Data, IEEE CG&A, Vol. 10(2), 1990;
[51] Goodsell, D., Mian, S., et al., Rendering Volumetric Data in Molecular Systems, J. Mol. Graphics, Vol. 7(3), 1989;
[52] von Walsum, T., Hin, A., et al., Efficient Hybrid Rendering of Volume Data And Polygons, Proc. 2nd Eurographics Workshop on ViSC, 1991;
[53] Fruhauf, M., Combining Volume Rendering with Line and Surface Rendering, Eurogriphics'91, 1991;
[2] 贺国珠，李显耀，毕会英，核磁共振技术应用的基本原理，物理通报，Vol．12(7)，1997；
[3] 巫北海，CT与MRI进展，第三军医大学学报，Vol．24(6)，2002：627-630；
[4] H.D William, The World Medical Equipment, Vol.12(10), 1999;
[5] Brink JA. Technical aspects of helical (spiral) CT [J], Radiol Clin North Am, Vol.33(4), 1995;
[6] 沈彬 李加 张彦军 李宁，新型光学成像技术及其在生物医学中的应用，激光杂志，Vol．20(2)，1999；
[7] 特集，最近的显微镜，精密工学会杂志，Vol．57(7)，1991；
[8] Sjostrand FS, J Ultrastrust, Res, Vol.2, 1958;
[9] 邓利群，快速制作常规石蜡切片，湖北民族学院学报(医学版)，Vol．18(4)，2001；
[10] 曹敏年，一种用PLC控制步进电机的方法，上海电器技术，Vol4(17)，1996；
[11] 范正翘 刘进 程胜，单片机控制的步进电动机综合微步驱动系统，微特电机，Vol3．，1996；
[12] 郁道银 谈恒英，光学工程，机械工业出版社，5月，1999；
[13] 李晓彤，几何光学与光学设计，浙江大学出版社，7月，1997；
[14] Noriniko Nakano, et al., Digital Still Camera System for Megapixel CCD, IEEE Transaction on Consumer Electronics, 44(3), 1998;
[15] 陆勤编译，[美]Popular Photograghy，数字相机地实拍评价，照相机，Vol6；1998；
[16] 石教英 蔡文立著，科学计算可视化算法与系统，科学出版社，9，1996；
[17] Cline, H., Lorensen, W., et al., Two Algorithms for the 3D Reconstruction of Tomograms, Medical Physics, Vol. 15(3), 1988;
[18] Farin, G.E,Curves and Surface From Computer Aided Geometric Design, Academic Press, 1990;
[19] Levoy, M., Display of surfaces from Volume Data, IEEE CG&A, Vol.8(5), 1990;
[20] Fruhauf, M., Combining Volume Rendering with Line and Surface Rendering, Eurogriphics'91, 1991;
[21] Shinagawa, Y., Kunii, T.L., Constructing a Reeb Graph Automatically from Cross Section, IEEE CG&A, Vol. 11,1991;
[22] Christiansen, H.N., Sederberg, T.W., Construction of Complex contours Line Definition into Polygonal Element Mosaics, Computer Graphics, Vo1.12(3), 1978;
[23] 沈兰荪，图像编码与异步传输。北京：人民邮电出版社，1998；
[24] 林靖 陈辉堂 蒋平 王月娟，用多媒体视频卡实现实时数字图像采集，测控技术，Vol．18(2)，1999；
[25] 王永刚，卫保国，基于数码相机的图像采集系统，测控技术，Vol．19 No．5，2000
[26] Norihiko Nakano, et al., Digital Still Camera System for Megapixel CCD. IEEE transaction on Consumer Electronics, Vol.44(3), 1998;
[27] Mendis S K, et al. Progress in CMOS active pixel image sensors. SPIE Proceedings, 2172,1995;
[28] 陆勤编译，[美]Popular Photography，数字相机的实拍评价，照相机，Vol．6，1998；
[29] 谭玉柱 侯晓轩，三相反应式步进电机驱动接口电路设计及应用，中小型电机，27(1)2000；
[30] 陈瑛等，在微机上实现脑结构三维重构技术，中国医学影像技术，1999，15(12)；
[31] 王今著，生物组织显微镜结构三维重建的灰度阴影立体图对显示技术，生物物理学报，2(3)17，1990；
[32] Heffeman PB, Robb RA. A new method of shaded surface display of biological and medical image. IEEE Trans on Medical Imaging, MI-4 25, 1985;
[33] Udapa JK. Interactive segmentation and boundary surface formation for 3D digital images. Comput Graph Imag Proc, 18(15)213, 1982;
[34] 李淑梁，陈洁．三维CT图像的显示与重建．清华大学学报，26(5)10，1986；
[35] 张义，李时光，杨恬．生物组织显微切片图像计算机三维重建的截面重建技术，中国生物医学工程学报，11(1)17，1992；
[36] 陈qing，多目标离散断层图像序列的三维重建算法研究，重庆大学硕士学位论文，1995；
[37] 吕凤军，数字图像处理编程入门，清华大学出版社，1999．9；
[38] 束越新．《颜色光学基础理论》．济南：山东科学技术出版社，47-64，1981．
[39] 刘维一等，用空间图像融合技术进行三维重建的研究，光学学报，20(3)，2000；
[40] Christiansen, H.N., Sederberg, T.W., Construction of Complex contours Line Definition into Polygonal Element Mosaics, Computer Graphics, 12(3), 1978;
[41] Fchis, H., Kendem, Z., et al.,Optimal Surface Reconstruction from planar Contours, CACM, Vol. 20, 1979;
[42] Christiansen, H.N., Sederberg, T.W., Construction of Complex Contours Line Definition into Polygonal Element Mosaics, Computer Graphics, Vol. 12(3), 1978;
[43] Cook, L.T., Dwyer, S., et al., A Three Dimensional Display System for Diagnostic Imaging Applications, IEEE CG&A, Vol. 3(4), 1983;
[44] Gananpathy, S., Dennely, T.G., A new General Triangulation Method for Planar Contours, Computer Graphics, Vol. 16(3), 1982;
[45] Ekoule, A.B., Peyrin, F.C., et al., A Triangulation Algorithm from Arbitrary Shaped Multiple Planar Contours, ACM Trans. On Graphics, Vol. 10, 1991;
[46] Jaffery, S., Dutta, K., Digital Reconstruction Methods for 3D Image Visualization, SPIE, Vol.507, 1984;
[47] Sabella, P., A rendering Algorithm for Visualization 3D Scalar Fields, Computer Graphics, Vol. 22(4), 1988;
[48] Drebin, R.L., Carpenter R., et al., Volume Rendering, Computer Graphics, Vol. 18(3), 1984;
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