Wilson Tony
Cold Spring Harb Protoc. 2010 Nov 1;2010(11):pdb.top88. doi: 10.1101/pdb.top88.
The popularity of the confocal microscope in life science laboratories around the world is undoubtedly due to its ability to permit volume objects to be imaged and to be rendered in three dimensions. It is important to realize that the confocal microscope itself does not produce three-dimensional images. Indeed, it does the opposite. The critical property that the confocal microscope possesses, which the conventional microscope does not, is its ability to image efficiently (and in-focus) only those regions of a volume specimen that lie within a thin section in the focal region of the microscope. In other words, it is able to reject (i.e., vastly attenuate) light originating from out-of-focus regions of the specimen. To image a three-dimensional volume of a thick specimen, it is necessary to take a whole series of such thin optical sections as the specimen is moved axially through the focal region. Once this through-focus series of optically sectioned images has been recorded, it is a matter of computer processing to decide how the three-dimensional information is to be presented. There are many methods for producing optical sections, of which the confocal optical system is just one. This article reviews these methods and describes a number of convenient methods of implementation that can lead to, among other things, real-time image formation.
共聚焦显微镜在世界各地生命科学实验室中的普及，无疑归功于其能够对立体物体进行成像并呈现三维图像的能力。必须认识到，共聚焦显微镜本身并不会产生三维图像。事实上，恰恰相反。共聚焦显微镜所具备而传统显微镜所没有的关键特性，是其仅能高效地（且聚焦清晰地）对位于显微镜焦区内薄切片中的立体标本区域进行成像的能力。换句话说，它能够抑制（即大幅衰减）来自标本失焦区域的光。要对厚标本的三维体积进行成像，当标本沿轴向穿过焦区时，有必要采集一整套这样的薄光学切片。一旦记录下这一通过聚焦的光学切片图像序列，如何呈现三维信息就只是计算机处理的问题了。产生光学切片的方法有很多种，共聚焦光学系统只是其中之一。本文将对这些方法进行综述，并描述一些便捷的实现方法，这些方法尤其能够实现实时图像形成。
