Weber Michael, Mickoleit Michaela, Huisken Jan
Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Dresden, Germany.
Methods Cell Biol. 2014;123:193-215. doi: 10.1016/B978-0-12-420138-5.00011-2.
This chapter introduces the concept of light sheet microscopy along with practical advice on how to design and build such an instrument. Selective plane illumination microscopy is presented as an alternative to confocal microscopy due to several superior features such as high-speed full-frame acquisition, minimal phototoxicity, and multiview sample rotation. Based on our experience over the last 10 years, we summarize the key concepts in light sheet microscopy, typical implementations, and successful applications. In particular, sample mounting for long time-lapse imaging and the resulting challenges in data processing are discussed in detail.
本章介绍了光片显微镜的概念,并提供了有关如何设计和构建此类仪器的实用建议。由于具有高速全帧采集、最小光毒性和多视图样本旋转等几个优越特性,选择性平面照明显微镜被作为共聚焦显微镜的一种替代方法呈现。基于我们过去十年的经验,我们总结了光片显微镜的关键概念、典型实现方式和成功应用。特别是,详细讨论了用于长时间延时成像的样本安装以及由此产生的数据处理挑战。