Abstract：
In this chapter, approaches to the image analysis of the choreography of the plant endoplasmic reticulum (ER) labeled with fluorescent fusion proteins (\"stars,\" if you wish) are presented. The approaches include the analyses of those parts of the ER that are attached through membrane contact sites to moving or non-moving partners (other \"stars\"). Image analysis is also used to understand the nature of the tubular polygonal network, the hallmark of this organelle, and how the polygons change over time due to tubule sliding or motion. Furthermore, the remodeling polygons of the ER interact with regions of fundamentally different topologies, the ER cisternae, and image analysis can be used to separate the tubules from the cisternae. ER cisternae, like polygons and tubules, can be motile or stationary. To study which parts are attached to non-moving partners, such as domains of the ER that form membrane contact sites with the plasma membrane/cell wall, an image analysis approach called persistency mapping has been used. To study the domains of the ER that move rapidly and stream through the cell, image analysis of optic flow has been used. However, optic flow approaches confuse the movement of the ER itself with the movement of proteins within the ER. As an overall measure of ER dynamics, optic flow approaches are of value, but their limitation as to what exactly is \"flowing\" needs to be specified. Finally, there are important imaging approaches that directly address the movement of fluorescent proteins within the ER lumen or in the membrane of the ER. Of these, fluorescence recovery after photobleaching (FRAP), inverse FRAP (iFRAP), and single particle tracking approaches are described.
摘要：
在这一章中,用荧光融合蛋白标记的植物内质网（ER）的编舞图像分析方法（“恒星，\“如果你愿意的话)被提出来。这些方法包括分析通过膜接触位点附着到移动或非移动伴侣（其他“星星”）的ER部分。图像分析还用于了解管状多边形网络的性质，这个细胞器的标志,以及由于细管滑动或运动，多边形如何随时间变化。此外，ER的重塑多边形与根本不同拓扑的区域相互作用，水箱，和图像分析可用于从池中分离小管。水箱，比如多边形和小管，可以是活动的或静止的。为了研究哪些部分附着在不移动的伙伴身上，例如与质膜/细胞壁形成膜接触位点的ER结构域，已经使用了一种称为持久性映射的图像分析方法。为了研究快速移动并流经细胞的ER域，光流的图像分析已被使用。然而,光流方法将ER本身的运动与ER内蛋白质的运动混淆。作为ER动态的总体度量，光流方法是有价值的，但他们的限制究竟是什么“流动”需要指定。最后,有重要的成像方法可以直接解决荧光蛋白在ER腔内或ER膜内的运动。其中,光漂白后的荧光恢复（FRAP），反FRAP(iFRAP),描述了单粒子跟踪方法。
