Plant and Microbial Biology Department, North Carolina State University, Raleigh, NC, United States.
Electrical and Computer Engineering Department, North Carolina State University, Raleigh, NC, United States.
Methods Cell Biol. 2020;160:405-418. doi: 10.1016/bs.mcb.2020.04.004. Epub 2020 May 30.
Imaging technologies have been used to understand plant genetic and developmental processes, from the dynamics of gene expression to tissue and organ morphogenesis. Although the field has advanced incredibly in recent years, gaps remain in identifying fine and dynamic spatiotemporal intervals of target processes, such as changes to gene expression in response to abiotic stresses. Lightsheet microscopy is a valuable tool for such studies due to its ability to perform long-term imaging at fine intervals of time and at low photo-toxicity of live vertically oriented seedlings. In this chapter, we describe a detailed method for preparing and imaging Arabidopsis thaliana seedlings for lightsheet microscopy via a Multi-Sample Imaging Growth Chamber (MAGIC), which allows simultaneous imaging of at least four samples. This method opens new avenues for acquiring imaging data at a high temporal resolution, which can be eventually probed to identify key regulatory time points and any spatial dependencies of target developmental processes.
成像技术已被用于理解植物遗传和发育过程，从基因表达的动态到组织和器官形态发生。尽管近年来该领域取得了令人难以置信的进展，但在识别目标过程的精细和动态时空间隔方面仍存在差距，例如基因表达对非生物胁迫的响应变化。光片显微镜由于能够在低光毒性下以精细的时间间隔对垂直取向的活体幼苗进行长期成像，因此是此类研究的一种有价值的工具。在本章中，我们描述了一种通过多样本成像生长室（MAGIC）准备和对拟南芥幼苗进行光片显微镜成像的详细方法，该方法允许同时对至少四个样本进行成像。这种方法为以高时间分辨率获取成像数据开辟了新途径，最终可以探测到关键的调控时间点和目标发育过程的任何空间依赖性。
