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Sub-Nanometer Resolution of Protein-DNA Interactions Enabled by pMINFLUX and Graphene

Oct 15, 2024
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About This Webinar
Superresolution microscopy has overcome photon-limited resolution with the combination of single-molecule localization and structured illumination such as by MINFLUX microscopy. Tinnefeld showcases pulsed-interleaved MINFLUX (pMINFLUX) which simplifys the switch between the excitation donuts and he additionally provides fluorescence lifetime information. MINFLUX can be combined with lifetime imaging for superresolved FRET and for co-tracking of single molecules in the nanometer range.

Combined with graphene energy transfer and DNA PAINT, isotropic resolution before 2 nm is shown on DNA origami model structures. But resolving multiple coordinates through localization methods requires the system to be static over the time period of the measurement, typically ranging from seconds to hours. For superresolution to contribute to dynamic structured biology, the challenge of obtaining robust insights amidst translational or rotational movements undermines certain benefits fluorescence provides for operating at physiologically relevant conditions. Graphene energy transfer with vertical nucleic acids (GETvNA), a new fluorescence-based method, provides Angstrom resolution to study biomolecular complexes involving nucleic acids under biologically relevant conditions. Besides the axial information provided by graphene energy transfer, GETvNA exploits the vertical orientation spontaneously adopted by double-stranded DNA segments bound to graphene by single-stranded DNA overhangs. Tinnefeld shares how it is possible with GETvNA to bend angles of distorted DNA structures and visualize protein diffusion with single basepair stepping.

*** This presentation premiered during the 2024 BioPhotonics Conference. For more information on Photonics Media conferences and summits, visit events.photonics.com

About the presenter

Philip TinnefeldPhilip Tinnefeld is professor for physical chemistry at The Ludwig Maximilian University of Munich. For more than 15 years, he has led an interdisciplinary research group in single-molecule spectroscopy, super-resolution microscopy, DNA nanotechnology, and biosensing with a focus on method development. Tinnefeld has contributed to breakthroughs in super-resolution microscopy with the techniques dSTORM, DNA-PAINT and pMINFLUX. Recently, he combined single-molecule detection with DNA nanotechnology to develop self-assembled functional devices. In addition to more than 190 peer-reviewed publications, he is involved in 10 patent applications and is initiator of GATTAquant GmbH.
MicroscopyBiophotonicssuperresolution microscopygraphene
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