Macro optical projection tomography for large scale 3D imaging of plant structures and gene activity.
Macro optical projection tomography for large scale 3D imaging of plant structures and gene activity.
复制标题
DOI：
10.1093/jxb/erw452
复制
发表时间：
2017-01-01
影响因子：
6.9
通讯作者：
Coen ES
中科院分区：
文献类型：
--
作者：
Lee KJI;Calder GM;Hindle CR;Newman JL;Robinson SN;Avondo JJHY;Coen ES
Macro OPT可捕捉高达60 mm × 45 mm的植物3D形态，实现植物-昆虫相互作用和基因活性的可视化。光学投影断层扫描（OPT）是一种用于可视化植物和动物基因活性的成熟方法。然而，传统的OPT的限制是，试样的上限尺寸限制排除其应用到更大的结构。为了解决这个问题，我们构建了一个宏版本，称为宏OPT（M-OPT）。我们将M-OPT应用于生长中的整个植物的基因活性的3D实时成像，并可视化高达60 mm高和45 mm深的大型光学透明植物和昆虫标本的结构形态。我们还展示了如何M-OPT可用于固定组织内的3D基因表达结构域成像，并在生长的年轻拟南芥全株克隆中可视化3D基因活性。M-OPT的另一个应用是可视化植物-昆虫相互作用。因此，M-OPT提供了一个有效的3D成像平台，可以在宏观尺度上研究基因活性，植物内部结构和植物-昆虫相互作用。
Macro OPT captures 3D morphologies of plants up to 60 mm by 45 mm, enabling visualisation of plant-insect interactions and gene activity. Optical projection tomography (OPT) is a well-established method for visualising gene activity in plants and animals. However, a limitation of conventional OPT is that the specimen upper size limit precludes its application to larger structures. To address this problem we constructed a macro version called Macro OPT (M-OPT). We apply M-OPT to 3D live imaging of gene activity in growing whole plants and to visualise structural morphology in large optically cleared plant and insect specimens up to 60 mm tall and 45 mm deep. We also show how M-OPT can be used to image gene expression domains in 3D within fixed tissue and to visualise gene activity in 3D in clones of growing young whole Arabidopsis plants. A further application of M-OPT is to visualise plant-insect interactions. Thus M-OPT provides an effective 3D imaging platform that allows the study of gene activity, internal plant structures and plant-insect interactions at a macroscopic scale.