Protocol to fluorescently stain vacuoles in Arabidopsis root cells
Year: 2024
Type: article
Abstract: Plant vacuoles are essential organelles that respond to developmental and environmental signals. Here, we present a protocol for staining the vacuole lumen in Arabidopsis root cells, enabling precise visualization of vacuolar dynamics. We describe steps for preparing plant material and staining with commonly used fluorescent dyes. We then detail procedures for visualizing vacuoles in the blue, green, and red emission spectra, allowing for their combined use with a variety of compatible fluorescent-tagged protein markers. For complete details on the use and execution of this protocol, please refer to Dubrovsky et al., 1 Fricker, 2 Bassil et al., 3 Grzam et al., 4 and Stefano et al. 5 • Analysis of vacuole morphology in Arabidopsis root cells • Protocol to stain vacuoles and combine with fluorescent protein detection • Reconstruction of 3D vacuoles and quantifications of volume and morphology Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Plant vacuoles are essential organelles that respond to developmental and environmental signals. Here, we present a protocol for staining the vacuole lumen in Arabidopsis root cells, enabling precise visualization of vacuolar dynamics. We describe steps for preparing plant material and staining with commonly used fluorescent dyes. We then detail procedures for visualizing vacuoles in the blue, green, and red emission spectra, allowing for their combined use with a variety of compatible fluorescent-tagged protein markers.
Source: STAR Protocols
Authors: Wylie Kristopher Tiernan, Graciela Veronica Castro, Kurtis Shipman, Cecilia Rodriguez-Furlan
Institution: Washington State University
Language: English
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,500
OpenAlex ID: W4405665261