Three-dimensional tracking of organ development in live plants based on plasma membrane dyes at single-cell resolution

Abstract

Plant developmental biology necessitates precise three-dimensional (3D) tracking of dynamic processes in live plants, and the 3D imaging technique in developmental bioimaging requires suitable fluorophores to achieve single-cell resolution imaging. Herein, we have designed a series of plasma membrane fluorescent dyes with a number of excellent properties and established a single-cell resolution imaging tool based on these dyes for three-dimensional imaging of various tissues and organs in living plants. The designed plasma membrane fluorescent dyes not only have the advantages of rapid wash-free staining, highly specific targeting, high brightness and high contrast imaging, ultralong imaging time and low biotoxicity, but also effectively avoid the autofluorescence interference of chlorophyll in cells, allowing for the development of a three-dimensional imaging approach of living plant organs with single-cell resolution. The three-dimensional histological structures of various organs of adult Arabidopsis thaliana, including roots, leaves, flowers, and fruits, were successfully reconstructed with single-cell resolution using this model plant. Furthermore, the 3D imaging method was employed to track the dynamic changes in tissue and organ morphology at the single-cell level during key plant developmental processes, including seed germination, root development, leaf growth, and anther development.

Keywords: Fluorescent dyes; Live imaging; Single-cell resolution; Three-dimensional imaging; Three-dimensional tracking.