Single-shot two-dimensional nano-size mapping of fluorescent molecules by ultrafast polarization anisotropy imaging

Peng Wang, Yogeshwar Nath Mishra, Florian J. Bauer, Murthy S. Gudipati and Lihong V. Wang ()
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Peng Wang: Mail Code 138-78
Yogeshwar Nath Mishra: Mail Code 138-78
Florian J. Bauer: Universität Erlangen-Nürnberg
Murthy S. Gudipati: 4800 Oak Grove Drive
Lihong V. Wang: Mail Code 138-78

Nature Communications, 2025, vol. 16, issue 1, 1-14

Abstract: Abstract Molecular size plays a crucial role in determining the physical, chemical, and biological properties of substances. However, traditional fluorescence polarization anisotropy methods struggle to capture fast transient processes or provide plane-specific details. To overcome these limitations, we introduce Compressed Ultrafast Planar Polarization Anisotropy Imaging (CUP2AI). This approach combines femtosecond laser-sheet illumination, molecular rotational diffusivity, and compressed sensing to enable real-time, non-invasive, wide-field anisotropy measurements in both liquid and gas phases. CUP2AI captures 2D molecular size mapping in a single acquisition, granting unprecedented insights into dynamic events across various excitation modes (i.e. both one- and two-photon) and environmental conditions. It enables mapping of molecular volume (500 ų–80,000 ų) and hydrodynamic diameter (10 Å–50 Å) based on anisotropy lifetimes. We imaged fluorescein-conjugated dextran in water and polycyclic aromatic hydrocarbons in flames. CUP2AI holds transformative potential for applications ranging from molecular biology and drug design to nanoparticle formation.

Date: 2025
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DOI: 10.1038/s41467-025-60072-1

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