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

Authors:

Laurell F. Kessler, Yunqing Li; Ashwin Balakrishnan, Mike Heilemann

Keywords:

stimulated emission depletion; STED; quantitative microscopy; molecular counting; EGFR; DNA origami

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.

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