Doumane Mehdi, Lionnet Claire, Bayle Vincent, Jaillais Yvon, Caillaud Marie-Cécile
Laboratoire Reproduction et Développement des Plantes, Univ Lyon, ENS de Lyon, Université Claude Bernard Lyon 1, CNRS, INRA, Lyon, France.
Lyon Bio-Image, Plateau Technique d'Imagerie/Microscopie (PLATIM), UMS3444/US8 BioSciences Gerland-Lyon Sud, ENS de Lyon, Lyon, France.
Bio Protoc. 2017 Apr 20;7(8). doi: 10.21769/BioProtoc.2245.
Here we describe a protocol that enables to automatically perform time-lapse imaging of growing root tips for several hours. Plants roots expressing fluorescent proteins or stained with dyes are imaged while they grow using automatic movement of the microscope stage that compensates for root growth and allows to follow a given region of the root over time. The protocol makes possible the image acquisition of multiple growing root tips, therefore increasing the number of recorded mitotic events in a given experiment. The protocol also allows the visualization of more than one fluorescent protein or dye simultaneously, using multiple channel acquisition. We particularly focus on imaging of cytokinesis in root tip meristem, but this protocol is also suitable to follow root hair growth, pollen tube growth, and other regions of root over time, in various plant species. It may as well be amendable to automatically track non-plant structures with an apical growth.
在此，我们描述了一种协议，该协议能够对生长中的根尖进行数小时的延时成像。表达荧光蛋白或用染料染色的植物根在生长过程中成像，通过显微镜载物台的自动移动来补偿根的生长，并允许随时间跟踪根的给定区域。该协议使对多个生长中的根尖进行图像采集成为可能，从而增加了给定实验中记录的有丝分裂事件的数量。该协议还允许使用多通道采集同时可视化多种荧光蛋白或染料。我们特别关注根尖分生组织中胞质分裂的成像，但该协议也适用于随时间跟踪各种植物物种的根毛生长、花粉管生长以及根的其他区域。它也可能适用于自动跟踪具有顶端生长的非植物结构。
