Quantitative Stimulated Emission Depletion (STED) Microscopy with DNA-Fluorophore Labels.

ABSTRACT: Stimulated emission depletion (STED) microscopy enables super-resolution imaging of complex biological samples in 3D, in large volumes, and live. However, molecular quantification with STED has remained underexplored. Here, we present a straightforward approach for quantitative STED that enables molecule counting. For this purpose, we designed DNA-fluorophore labels that enable signal amplification and allow for reliable intensity-based quantitative imaging. We demonstrate accurate molecule counting on DNA origami. Furthermore, we visualized and quantified EGF receptor monomers and dimers in cells. In summary, we introduce a robust, fast, and easy-to-implement tool for quantitative STED microscopy with single-protein resolution.

SUBMITTER: Kessler LF 

PROVIDER: S-EPMC12918720 | biostudies-literature | 2026 Feb

REPOSITORIES: biostudies-literature