Stimulated emission depletion (STED) microscopy is a powerful super-resolution microscopy technique that has achieved significant results in breaking the resolution limit of conventional confocal microscopes. In recent years, STED technology has continued to evolve, in particular with the aim of reducing the intensity of depletion lasers. In this workshop, we will use a STED microscope that has been combined with a fluorescence lifetime equipment, a combo known as lifetime STED microscopy. A phasor approach is used to improve the photon screening based on their lifetime and so to improve the resolution with less amount of depletion light. We applied this lifetime STED technique to plant tissue (Arabidopsis thaliana root tip) with the aim of imaging the organization of the Golgi apparatus at super-resolution and more precisely challenge the hypothetical presence of an Endoplasmic Reticulum (ER)-Golgi Intermediate Compartment (ERGIC) in plant cells. Stunningly, lifetime STED microscopy revealed the presence of ERGIC by clearly differentiating a thin tubulo-vesicular structure from the background noise. Moreover, we successfully performed Lifetime STED in two-colors (for Alexa 594 and ATTO647N) and visualized that this tubulo-vesicular network is mostly independent from either the medial-cisternae of the Golgi apparatus or the ER-exit sites (ERES), although some associations are clearly visible with these structures. Together, the lifetime STED approach allowed us to unravel a structure that remained previously unidentified in plants and thus represents a major step forward the understanding of Golgi organization in plant cells.