An automated single-molecule FRET platform for high-content, multiwell plate screening of biomolecular conformations and dynamics

Research output: Contribution to journalResearch articleContributedpeer-review


Single-molecule FRET (smFRET) has become a versatile tool for probing the structure and functional dynamics of biomolecular systems, and is extensively used to address questions ranging from biomolecular folding to drug discovery. Confocal smFRET measurements are amongst the widely used smFRET assays and are typically performed in a single-well format. Thus, sampling of many experimental parameters is laborious and time consuming. To address this challenge, we extend here the capabilities of confocal smFRET beyond single-well measurements by integrating a multiwell plate functionality to allow for continuous and automated smFRET measurements. We demonstrate the broad applicability of the multiwell plate assay towards DNA hairpin dynamics, protein folding, competitive and cooperative protein–DNA interactions, and drug-discovery, revealing insights that would be very difficult to achieve with conventional single-well format measurements. For the adaptation into existing instrumentations, we provide a detailed guide and open-source acquisition and analysis software.


Original languageEnglish
Article number6511
Number of pages12
JournalNature communications
Issue number1
Publication statusPublished - Dec 2023

External IDs

PubMed 37845199
ORCID /0000-0002-6209-2364/work/150328622
ORCID /0000-0002-2213-2763/work/150329031
ORCID /0000-0003-2125-4045/work/150329793
ORCID /0000-0002-4657-9092/work/150329890



  • Fluorescence Resonance Energy Transfer, Molecular Conformation, DNA, Software, Protein Folding

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