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Updated: Jul 14, 2026
Spectroscopic Super-resolution Imaging of DNA Molecules using Intrinsic Contrast
Published on: March 6, 2026
Single-pair fluorescence resonance energy transfer of nucleosomes in free diffusion: optimizing stability and
Alexander Gansen1, Florian Hauger, Katalin Tóth
1Division of Biophysics of Macromolecules (B040), German Cancer Research Center, D-69120 Heidelberg, Germany.
Abstract:
We applied fluorescence detection methods on the single-molecule level to study structural variations and dynamic processes occurring within nucleosomes. Four fluorescent nucleosome constructs were made by attaching donor and acceptor fluorophores to different positions of two nucleosome positioning sequences and reconstituting nucleosomes by salt dialysis. The photochemical and biochemical stability of nucleosomes under single-molecule conditions was optimized by adding inert protein and free radical capturing additives, allowing us to define the best experimental conditions for single-molecule spectroscopy on highly diluted solutions of nucleosome complexes. We could demonstrate for the first time the resolution of conformational subpopulations of nucleosomes by single-pair fluorescence resonance energy transfer in a freely diffusing system and could show the effect of thermally induced nucleosome repositioning.
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