Rasnik Ivan, French Todd, Jacobson Ken, Berland Keith
Physics Department, Emory University, Atlanta, Georgia, USA.
Methods Cell Biol. 2013;114:211-41. doi: 10.1016/B978-0-12-407761-4.00010-5.
This chapter introduces to electronic cameras, discusses the various parameters considered for evaluating their performance, and describes some of the key features of different camera formats. The chapter also presents the basic understanding of functioning of the electronic cameras and how these properties can be exploited to optimize image quality under low-light conditions. Although there are many types of cameras available for microscopy, the most reliable type is the charge-coupled device (CCD) camera, which remains preferred for high-performance systems. If time resolution and frame rate are of no concern, slow-scan CCDs certainly offer the best available performance, both in terms of the signal-to-noise ratio and their spatial resolution. Slow-scan cameras are thus the first choice for experiments using fixed specimens such as measurements using immune fluorescence and fluorescence in situ hybridization. However, if video rate imaging is required, one need not evaluate slow-scan CCD cameras. A very basic video CCD may suffice if samples are heavily labeled or are not perturbed by high intensity illumination. When video rate imaging is required for very dim specimens, the electron multiplying CCD camera is probably the most appropriate at this technological stage. Intensified CCDs provide a unique tool for applications in which high-speed gating is required. The variable integration time video cameras are very attractive options if one needs to acquire images at video rate acquisition, as well as with longer integration times for less bright samples. This flexibility can facilitate many diverse applications with highly varied light levels.
本章介绍电子相机,讨论评估其性能时考虑的各种参数,并描述不同相机格式的一些关键特性。本章还介绍了电子相机功能的基本理解,以及如何利用这些特性在低光条件下优化图像质量。尽管有许多类型的相机可用于显微镜检查,但最可靠的类型是电荷耦合器件(CCD)相机,在高性能系统中它仍然是首选。如果时间分辨率和帧率无关紧要,慢扫描CCD在信噪比和空间分辨率方面肯定能提供最佳性能。因此,慢扫描相机是使用固定标本的实验(如免疫荧光测量和荧光原位杂交)的首选。然而,如果需要视频速率成像,则无需评估慢扫描CCD相机。如果样品标记强烈或不受高强度照明干扰,一个非常基本的视频CCD可能就足够了。当非常暗的标本需要视频速率成像时,在这个技术阶段电子倍增CCD相机可能是最合适的。增强型CCD为需要高速选通的应用提供了一种独特的工具。如果需要以视频速率采集图像,同时对于不太亮的样品需要更长的积分时间,可变积分时间摄像机是非常有吸引力的选择。这种灵活性可以促进许多具有高度不同光照水平的多样化应用。