Shen Wei, Shen Yun, Lam Yun Wah, Chan Danny
Department of Biology and Chemistry, City University of Hong Kong, Kowloon Tong, Hong Kong.
School of Biomedical Sciences, The University of Hong Kong, Pokfulam, Hong Kong.
Methods Mol Biol. 2018;1774:507-518. doi: 10.1007/978-1-4939-7802-1_22.
Planarian regeneration involves a complex series of cellular events, precisely choreographed in space and time. Time-lapse imaging can provide powerful insights into tissue dynamics, as variously demonstrated in other model systems. However, time-lapse imaging of planarians has proven to be a challenge. Especially the requisite immobilization of the animals over extended periods of time is difficult, owing to their photophobic behavior and soft body architecture. Here, we describe a new embedding method using 2% (w/v) low melting agarose, and demonstrate that this method can effectively immobilize animals as long as 7 days. In combination with cell-permeable fluorescent dyes, this immobilization method allows for the time-lapse imaging of planaria during regeneration and other physiological processes.
涡虫再生涉及一系列复杂的细胞事件,这些事件在空间和时间上精确编排。延时成像可以深入了解组织动态,其他模型系统也有各种相关证明。然而,涡虫的延时成像已被证明是一项挑战。特别是由于它们的避光行为和柔软的身体结构,长时间固定这些动物很困难。在这里,我们描述了一种使用2%(w/v)低熔点琼脂糖的新包埋方法,并证明该方法可以有效固定动物长达7天。结合细胞可渗透的荧光染料,这种固定方法能够对涡虫再生及其他生理过程进行延时成像。