Biophysical Society Thematic Meeting | Tutzing 2026

Single-Molecule FRET: The Next 30 Years

Tuesday Speaker Abstracts

METHODOLOGIES FOR FLUORESCENCE BASED STRUCTURAL BIOLOGY OF COMPLEX SYSTEMS Jelle Hendrix 1 ; 1 Hasselt University, Hasselt, Belgium Biomolecular function is inherently dynamic, and resolving how structural fluctuations shape activity requires experimental approaches capable of capturing motion across molecular, spatial, and temporal scales. We employ single-molecule FRET (smFRET) to detect rapid conformational changes within individual molecules, even when state switching occurs during millisecond bursts, providing direct access to kinetic information. A major focus is the development of physiologically relevant model environments. Using GUVs, supported bilayers, and pore-spanning membranes, combined with two-photon polymerization, we fabricate custom 3D microstructures compatible with confocal and multiphoton microscopes. Microfluidic devices enable controlled flow, droplet manipulation, and long-duration smFRET, supported by array-detector FCS and 3D flow-velocity mapping. Finally, resistive microheating and lock-in detection allow precise thermal perturbations for extracting kinetic parameters, while solvatochromic dyes (e.g., ACDAN) report on local environments through spectral and lifetime phasors. Together, these methods form a versatile, microscope-integrated toolkit for quantifying biomolecular dynamics in tunable microenvironments.

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