Advances in Synthetic Fluorescent Probe Labeling for Live-Cell Imaging in Plants

Abstract

Fluorescent probes are a powerful tool for visualizing cellular and subcellular structures, their dynamics, and cellular molecules in living cells, and enables us to monitor cellular processes in a spatio-temporal manner within complex and crowded systems. In addition to popular fluorescent proteins, a wide variety of small-molecule dyes have been synthesized through close association with the interdisciplinary field of chemistry and biology, ranging from those suitable for labeling cellular compartments such as organelles to those for labeling intracellular biochemical and biophysical processes and signaling. In recent years, self-labeling technologies including the SNAP-tag system have allowed us to attach these dyes to cellular domains or specific proteins, and are beginning to be employed in plant studies. In this mini review, we will discuss the current range of synthetic fluorescent probes that have been exploited for live cell imaging and the recent advances in the application that enable genetical tagging of synthetic probes in plant research.

A versatile CRISPR-based system for lineage tracing in living plants

“…Still, they require tissue fixation, which is incompatible with observation in living organisms. This limit is overcome by using fluorescent proteins (FPs), with manifold advantages (Yagi et al 2021).…”

“…Still, they require tissue fixation, which is incompatible with observation in living organisms. This limit is overcome by using fluorescent proteins (FPs), with manifold advantages (Yagi et al 2021). FPs are encoded by relatively short genes, making transgene design easier, and they can be fused as tags to other proteins, often without functional interference.…”

Long-term spatiotemporal and highly specific imaging of the plasma membrane of diverse plant cells using a near-infrared AIE probe

“…Biologists generally track and monitor the endocytosis process in living cells using specific fluorescent proteins on the plasma membrane, but fluorescent proteins have also shown several disadvantages such as cumbersome operation and low photostability. The fluorescent proteins currently known are nearly always accompanied by a significant decrease in brightness during long-term fluorescence imaging, 4,5 incomplete fluorophore maturation of fluorescent proteins frequently impairs signal readouts and their large size also restricts functionality for site-specific labeling. 6 In addition to fluorescent proteins, scientists working in chemistry and biology and have developed a variety of small molecular dyes which can be used as markers of diverse subcellular organelles and track various biochemical and biophysical processes of cells.…”

A versatile CRISPR‐based system for lineage tracing in living plants

“…Still, they require tissue fixation, which is incompatible with observation in living organisms (see for example Dolan et al., 1994; Scheres et al., 1994). This limit is overcome by using fluorescent proteins (FPs), with manifold advantages (Yagi et al., 2021). FPs are encoded by relatively short genes, making transgene design easier, and they can be fused as tags to other proteins, often without functional interference.…”