# Dynamic Binder Exchange Improves Protein Labeling Efficiency in DNA‐PAINT up to 15‐Fold

## TL;DR

A new method called DyBE improves DNA-PAINT microscopy by 15 times, allowing clearer imaging of protein structures at the nanoscale.

## Contribution

DyBE introduces a dual-kinetic scheme using transient binder-target interactions to boost labeling efficiency in super-resolution imaging.

## Key findings

- DyBE increases labeling efficiency up to 15-fold in DNA-PAINT microscopy.
- DyBE enables detection of HER2 homodimers and EGFR-HER2 heterodimers at single-protein resolution.
- The method expands the range of usable binders for spatial proteomics and drug studies.

## Abstract

Dynamic Binder Exchange (DyBE) enhances DNA‐PAINT (Point Accumulation for Imaging in Nanoscale Topography) super‐resolution microscopy by exploiting transient, reversible binder‐target interactions. DyBE uses DNA‐conjugated binders such as nanobodies as both targeting and docking moieties, integrating their characteristic higher off‐rates with DNA‐PAINT blinking to efficiently sample target sites. This dual‐kinetic scheme increases labeling efficiency up to 15‐fold, enabling sensitive detection of targets previously inaccessible due to limitations of high‐off‐rate binders. Using DyBE, the study reveals pre‐existing HER2 homodimers and ligand‐induced EGFR‐HER2 heterodimers at single‐protein resolution with high fidelity. DyBE expands the usable binder repertoire, advancing spatial proteomics and enabling mechanistic drug studies of receptor organization and signaling.

Dynamic Binder Exchange (DyBE) leverages transient binder–target interactions to enhance labeling efficiency by up to 15‐fold in super‐resolution microscopy. DyBE maps nanoscale receptor tyrosine kinase (RTK) assemblies, including HER2 homodimers and EGFR–HER2 heterodimers, advancing single‐protein spatial proteomics.