New DNA-based localisation microscopy approaches to study protein-protein interactions and nanoscale protein placement in situ

Alex Clowsley; William T. Kaufhold; Tobias Lutz; Anna Meletiou; Lorenzo Di Michele; Christian Soeller

doi:10.1117/12.2584755

4 October 2020 New DNA-based localisation microscopy approaches to study protein-protein interactions and nanoscale protein placement in situ

Alex Clowsley, William T. Kaufhold, Tobias Lutz, Anna Meletiou, Lorenzo Di Michele, Christian Soeller

Author Affiliations +

Proceedings Volume 11575, Biophotonics and Biomedical Microscopy; 1157507 (2020) https://doi.org/10.1117/12.2584755
Event: SPIE Photonex and Vacuum Expo, 2020, Online Only

Abstract

It is crucial to visualize biomolecules and their molecular-interaction complexes directly within cells, to show precisely where these interactions occur and thus improve our understanding of cellular regulation. In this presentation I will present new approaches to image proteins and protein-protein interactions that are based on synthetic DNA interactions and allow localisation with ~10 nm precision. Two schemes will be presented, called PD-PAINT (proximity-dependent PAINT) and Repeat DNA-PAINT which are improvements of the localisation microscopy technique called DNA-PAINT. I will describe the basis of these approaches and demonstrate their application to biophysical questions in cardiac biophysics. We show that these schemes can be straightforwardly integrated in a multiplexed super-resolution imaging protocol and benefit from advantages of DNA-based super-resolution localization microscopy, such as high specificity, high resolution and the ability to image quantitatively.

Conference Presentation

Citation Download Citation

Alex Clowsley, William T. Kaufhold, Tobias Lutz, Anna Meletiou, Lorenzo Di Michele, and Christian Soeller "New DNA-based localisation microscopy approaches to study protein-protein interactions and nanoscale protein placement in situ", Proc. SPIE 11575, Biophotonics and Biomedical Microscopy, 1157507 (4 October 2020); https://doi.org/10.1117/12.2584755

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