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<div class="csl-entry">Gaugutz, A., Velas, L., Georgiou, E., Xing, Y., Moser, S., Maloberti Julian, Jesacher, A., Howorka, S., & Schütz, G. (2025, July 3). <i>3D Single Molecule Tracking of Membrane-Bound Proteins and DNA Nanoprobes using Defocused Imaging</i> [Poster Presentation]. 15th European Biophysics Congress (EBSA 2025), Rom, Italy. http://hdl.handle.net/20.500.12708/217197</div>
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dc.identifier.uri
http://hdl.handle.net/20.500.12708/217197
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dc.description.abstract
Protein movement and biomolecular interactions at cellular interfaces are strongly influenced by the nanoscale topography of biological membranes. Tracking individual membrane proteins within this complex cellular 3D environment at high spatial resolution is essential to understand biological mechanisms. While standard single molecule localization microscopy (SMLM) provides high localisation precision in lateral dimensions, its axial performance is limited. To achieve high localisation precision also in axial dimension, we present a 3D SMLM approach which uses defocused imaging to track single molecules in all three dimensions with an axial resolution down to 11 nm, as demonstrated with two fluorophore-tagged DNA nanoprobes on supported lipid bilayers. We also follow the 3D motion of T cell receptors in live cells, capturing their diffusion characteristics in activating and resting conditions. Our results lay the foundation for high-resolution 3D tracking of molecules, enabling new insights into the spatial organisation and dynamics of cellular processes.
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dc.language.iso
en
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dc.subject
DNA Origami
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dc.subject
Single Molecules
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dc.subject
Fluorescence Microscopy
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dc.title
3D Single Molecule Tracking of Membrane-Bound Proteins and DNA Nanoprobes using Defocused Imaging
