Dual-color DNA-PAINT single-particle tracking enables extended studies of membrane protein interactions

DNA-PAINT based single-particle tracking (DNA-PAINT-SPT) has recently significantly enhanced observation times in in vitro SPT experiments by overcoming the constraints of fluorophore photobleaching. However, with the reported implementation, only a single target can be imaged and the technique cann...

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Bibliographic Details
Published in:Nature communications 2023-07, Vol.14 (1), p.4345-4345, Article 4345
Main Authors: Niederauer, Christian, Nguyen, Chikim, Wang-Henderson, Miles, Stein, Johannes, Strauss, Sebastian, Cumberworth, Alexander, Stehr, Florian, Jungmann, Ralf, Schwille, Petra, Ganzinger, Kristina A.
Format: Article
Language:English
Subjects:
631/1647/245/2225
631/1647/328/2239
631/57/2265
Cell Membrane - metabolism
Cell membranes
Color
Deoxyribonucleic acid
Dimerization
DNA
DNA - metabolism
Dyes
Experiments
Humanities and Social Sciences
Labeling
Ligands
Localization
Membrane proteins
Membrane Proteins - metabolism
Membranes
multidisciplinary
Particle tracking
Photobleaching
Photochemical reactions
Protein interaction
Proteins
Science
Science (multidisciplinary)
Single Molecule Imaging - methods
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Summary:DNA-PAINT based single-particle tracking (DNA-PAINT-SPT) has recently significantly enhanced observation times in in vitro SPT experiments by overcoming the constraints of fluorophore photobleaching. However, with the reported implementation, only a single target can be imaged and the technique cannot be applied straight to live cell imaging. Here we report on leveraging this technique from a proof-of-principle implementation to a useful tool for the SPT community by introducing simultaneous live cell dual-color DNA-PAINT-SPT for quantifying protein dimerization and tracking proteins in living cell membranes, demonstrating its improved performance over single-dye SPT. Single-particle tracking (SPT) has revolutionised studies of protein interactions but is often limited by photobleaching. Here, the authors evolve DNA-PAINT-SPT to enable simultaneous dual-colour detection for the quantification of protein dimerization and live cell membrane protein tracking.
ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-023-40065-8