Deconvolution microscopy
Deconvolution microscopy
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DOI：
10.1007/b102215
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发表时间：
2005-01-01
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影响因子：
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通讯作者：
Sibarita, JB
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文献类型：
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作者：
Sibarita, JB
自 1983 年推出以来，反卷积显微镜已成为一种关键的图像处理工具，用于在三维和亚分辨率尺度上可视化固定和活体标本的细胞结构。过去 20 年见证了许多基于反卷积显微镜的不同应用的发展，包括各种光学设置和反卷积算法。本章旨在从理论方面到实际解决方案总结并详细描述该技术的主要特征。首先将解释三维光学切片显微镜中的图像形成原理。由于反卷积显微镜本质上提供了一种克服光学显微镜局限性的方法，因此本章的第二部分致力于通过显微镜生成图像的理论和实验描述。将详细介绍确定点扩散函数的方法，这是表征任何光学系统的关键步骤，也是图像反卷积的关键初步步骤。将讨论所面临的挑战以及精确确定该函数的各种可能性。将讨论所有可能的像差来源和图像退化过程。在本章的第三部分，我们将介绍采集设置以及采集和反卷积过程之间的一致性要求。将详细介绍固定细胞和活细胞观察的典型设置，以及优化速度和减少伪影的关键功能。在本章的第四部分和最后一部分中，我们将从理论上描述光学显微镜领域常用的各种恢复算法，并将提供使用一些商用软件包获得的结果。最后，我们将考虑未来解决方案（目前正在开发）的前景，旨在更轻松地处理快速多维活细胞显微镜生成的大量数据。本章专为标准细胞生物学家以及硬件和软件工程师和开发人员使用而设计，旨在为反卷积显微镜的广泛而强大的领域提供清晰的解释。
Since its introduction in 1983, deconvolution microscopy has become a key imageprocessing tool for visualizing the cellular structures of fixed and living specimens in three dimensions and at subresolution scale. The last 20 years have seen the development of many different applications based on deconvolution microscopy, including a wide variety of optical setup and deconvolution algorithms. This chapter aims to summarize and to describe in detail the major features of this technology, from theoretical aspects to practical solutions. It will begin by explaining the principle of image formation in three-dimensional optical sectioning microscopy. As deconvolution microscopy provides, in essence, a means of overcoming the limits of optical microscopy, the second part of this chapter is dedicated to the theoretical and experimental description of image generation through a microscope. Methods will be detailed for the determination of point spread function, as a crucial step for the characterization of any optical system and a key preliminary step for image deconvolution. The challenges faced and the various possibilities for determining this function precisely will be discussed. All possible sources of aberrations and image degradation processes will be discussed. In the third part of this chapter, we will introduce the acquisition setup and requirements for compliance between acquisition and deconvolution processes. Typical setups for fixed and living cell observation will be detailed, with key features for optimizing speed and reducing artifacts. in the fourth and last part of this chapter, we win describe, in theoretical terms, the various restoration algorithms commonly used in the field of optical microscopy and will provide results obtained with some of the commercially available packages. We shall conclude by considering the prospects for future solutions (currently under development) aiming to handle more easily the huge amounts of data generated by rapid multi-dimensional living cell microscopy. Designed for use by standard cell biologists and hardware and software engineers and developers, this chapter has been written to provide a clear explanation of the wide-reaching and powerful domain of deconvolution microscopy.