酵母细胞器的4D共聚焦成像
4D Confocal Imaging of Yeast Organelles.
作者信息
Day Kasey J, Papanikou Effrosyni, Glick Benjamin S
机构信息
Department of Molecular Genetics and Cell Biology, University of Chicago, 920 East 58th St., Chicago, IL, 60637, USA.
出版信息
Methods Mol Biol. 2016;1496:1-11. doi: 10.1007/978-1-4939-6463-5_1.
Abstract
Yeast cells are well suited to visualizing organelles by 4D confocal microscopy. Typically, one or more cellular compartments are labeled with a fluorescent protein or dye, and a stack of confocal sections spanning the entire cell volume is captured every few seconds. Under appropriate conditions, organelle dynamics can be observed for many minutes with only limited photobleaching. Images are captured at a relatively low signal-to-noise ratio and are subsequently processed to generate movies that can be analyzed and quantified. Here, we describe methods for acquiring and processing 4D data using conventional scanning confocal microscopy.
摘要
酵母细胞非常适合通过四维共聚焦显微镜观察细胞器。通常，一个或多个细胞区室用荧光蛋白或染料标记，每隔几秒就采集一叠跨越整个细胞体积的共聚焦切片。在适当条件下，仅需有限的光漂白就能观察细胞器动态达数分钟之久。图像以相对较低的信噪比采集，随后进行处理以生成可分析和定量的影片。在这里，我们描述了使用传统扫描共聚焦显微镜获取和处理四维数据的方法。
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