We report the development of HoT autoblinking DNA probes for quantitative
superresolution imaging of chromatin in living cells and tissues. Combining high cell
permeability, spontaneous blinking and superior photophysical performance, HoT
probes enable wash-free, additive-free single molecule localization microscopy
without genetic manipulation. They achieve localization precision around 20 nm by
2D STORM and down to ~3 nm in 2D MINFLUX, allowing visualization of
nanoscale chromatin architecture, DNA fiber dynamics, and nucleosomescale
organization.We demonstrate their utility for long-term real-time visualization of
chromatin architecture and dynamics, 3D nanoscale chromatin, quantitative chromatin
compaction measurement in tissue sections, combined OligoSTORM imaging of
individual genomic loci as well as 3D MINFLUX. HoTs enable quantitative
chromatin analysis in diverse models—from cultured cells and stem cells to zebrafish
retinal sections and human cancer tissue specimens—offering a powerful tool for both
basic chromatin biology and clinical diagnostics.This was a collaborative work with
the lab of Hongyan Sun (City U).
Wang et al. Molecular Cell, 2026
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