Bayguinov Peter O, Oakley Dennis M, Shih Chien-Cheng, Geanon Daniel J, Joens Matthew S, Fitzpatrick James A J
Center for Cellular Imaging, Washington University in St. Louis, St. Louis, Missouri.
Departments of Cell Biology & Physiology and Neuroscience, Washington University School of Medicine, St. Louis, Missouri.
Curr Protoc Cytom. 2018 Jul;85(1):e39. doi: 10.1002/cpcy.39. Epub 2018 Jun 20.
Since its commercialization in the late 1980's, confocal laser scanning microscopy (CLSM) has since become one of the most prevalent fluorescence microscopy techniques for three-dimensional structural studies of biological cells and tissues. The flexibility of the approach has enabled its application in a diverse array of studies, from the fast imaging of dynamic processes in living cells, to meticulous morphological analyses of tissues, and co-localization of protein expression patterns. In this chapter, we introduce the principles of confocal microscopy and discuss how the approach has become a mainstay in the biological sciences. We describe the components of a CLSM system and assess how modern implementations of the approach have further expanded the use of the technique. Finally, we briefly outline some practical considerations to take into account when acquiring data using a CLSM system. © 2018 by John Wiley & Sons, Inc.
自20世纪80年代末商业化以来,共聚焦激光扫描显微镜(CLSM)已成为用于生物细胞和组织三维结构研究的最普遍的荧光显微镜技术之一。该方法的灵活性使其能够应用于各种各样的研究中,从活细胞动态过程的快速成像,到组织的细致形态分析,以及蛋白质表达模式的共定位。在本章中,我们介绍共聚焦显微镜的原理,并讨论该方法如何成为生物科学的支柱。我们描述CLSM系统的组件,并评估该方法的现代应用如何进一步扩展了该技术的用途。最后,我们简要概述在使用CLSM系统获取数据时需要考虑的一些实际问题。© 2018 John Wiley & Sons, Inc.