Dual Color tau-STED Super Resolution Microscopy in Arabidopsis Root Tip
Abstract
Super-resolution microscopy has transformed our ability to visualize subcellular structures, but its application in plant biology remains challenging due to the optical complexity of plant tissues. Here, we present a detailed protocol for tau-STED microscopy (Leica Microsystems), which combines stimulated emission depletion (STED) with fluorescence lifetime imaging (FLIM) to achieve nanoscale resolution while minimizing phototoxicity. This method leverages time-correlated single-photon counting (TCSPC) to separate fluorescence signals based on their lifetimes, enhancing signal specificity and enabling the visualization of elusive subcellular compartments in Arabidopsis thaliana root tips. The protocol covers sample preparation, fluorophore selection, microscope configuration, image acquisition, and data analysis, providing a step-by-step guide to optimize tau-STED imaging for plant cell biology. By addressing the unique challenges of plant tissue imaging, such as autofluorescence, refractive index mismatches, and light scattering, this approach facilitates super-resolution imaging of intracellular structures, including the plant endoplasmic reticulum-Golgi intermediate compartment (ERGIC). This protocol is designed to be accessible to researchers with basic microscopy experience and offers a robust framework for exploring subcellular dynamics in plants with unprecedented detail. Key features • tau-STED integrates STED signals with fluorescence lifetime via phasor analysis at confocal speeds, enabling low-noise super-resolution imaging. • Morphometry analysis workflow at super resolution.
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BibTeX
@article{fougere2026dual,
title = {{Dual Color tau-STED Super Resolution Microscopy in Arabidopsis Root Tip}},
author = {Louise Fougère and Christel Poujol and Yohann Boutté and Magali Grison},
journal = {Bio-protocol},
year = {2026},
month = aug,
doi = {10.21769/bioprotoc.5774},
url = {https://hal.science/hal-05843896v1},
note = {CC BY-NC 4.0},
}
RIS
TY - JOUR
TI - Dual Color tau-STED Super Resolution Microscopy in Arabidopsis Root Tip
AU - Louise Fougère
AU - Christel Poujol
AU - Yohann Boutté
AU - Magali Grison
PY - 2026
DA - 2026/08/05
JO - Bio-protocol
DO - 10.21769/bioprotoc.5774
UR - https://hal.science/hal-05843896v1
AB - Super-resolution microscopy has transformed our ability to visualize subcellular structures, but its application in plant biology remains challenging due to the optical complexity of plant tissues. Here, we present a detailed protocol for tau-STED microscopy (Leica Microsystems), which combines stimulated emission depletion (STED) with fluorescence lifetime imaging (FLIM) to achieve nanoscale resolution while minimizing phototoxicity. This method leverages time-correlated single-photon counting (TCSPC) to separate fluorescence signals based on their lifetimes, enhancing signal specificity and enabling the visualization of elusive subcellular compartments in Arabidopsis thaliana root tips. The protocol covers sample preparation, fluorophore selection, microscope configuration, image acquisition, and data analysis, providing a step-by-step guide to optimize tau-STED imaging for plant cell biology. By addressing the unique challenges of plant tissue imaging, such as autofluorescence, refractive index mismatches, and light scattering, this approach facilitates super-resolution imaging of intracellular structures, including the plant endoplasmic reticulum-Golgi intermediate compartment (ERGIC). This protocol is designed to be accessible to researchers with basic microscopy experience and offers a robust framework for exploring subcellular dynamics in plants with unprecedented detail. Key features • tau-STED integrates STED signals with fluorescence lifetime via phasor analysis at confocal speeds, enabling low-noise super-resolution imaging. • Morphometry analysis workflow at super resolution.
SN - 2331-8325
ER -