Sequential replicas for in vivo imaging of growing organ surfaces

Sequential replica method facilitates in vivo imaging of plant surface and provides data sufficient for detailed computation of geometry and growth. It enables obtaining a series of high-resolution images visualizing details of the examined surface. Series of molds, made in dental polymer, representing the examined surface are used to obtain casts in epoxy resin, which are in turn observed by scanning electron microscopy, while the structure itself remains intact. Images obtained from casts can be further used for data extraction, comprising 3D reconstruction and computation of local geometry and cell growth parameters. The sequential replica method is a universal method and can be applied to image complex shapes of a range of structures, like meristems, flowers, stems, leaves, or various types of trichomes. Different plant species growing in various conditions can be studied.