Abstract：
In this chapter, I present a method for the ex vivo cultivation and live imaging of Drosophila imaginal disc explants using low concentrations of the steroid hormone 20-hydroxyecdysone (20E). This method has been optimized for analyzing cellular dynamics during wing disc growth and leverages recent insights from in vivo experiments demonstrating that 20E is required for growth and patterning of the imaginal tissues. Using this protocol, we directly observe wing disc proliferation at a rapid rate for at least 13 h during live imaging. The orientation of tissue growth is also consistent with that inferred from indirect in vivo techniques. Thus, this method provides an improved way of studying dynamic cellular processes and tissue movements during imaginal disc development. I first describe the preparation of the growth medium and the dissection, and then I include a protocol for mounting and live imaging of the explants.
摘要：
在这一章中,我提供了一种使用低浓度的类固醇激素20-羟基蜕皮激素（20E）对果蝇视盘外植体进行离体培养和实时成像的方法。该方法已经过优化，用于分析翼盘生长过程中的细胞动力学，并利用了来自体内实验的最新见解，证明20E是假想组织的生长和图案化所必需的。使用这个协议，在实时成像过程中，我们直接观察翼盘的快速增殖至少13小时。组织生长的方向也与从间接体内技术推断的方向一致。因此，这种方法为研究视盘发育过程中的动态细胞过程和组织运动提供了一种改进的方法。我首先描述了生长培养基的制备和解剖，然后我包括一个安装和实时成像的外植体的协议。
