DNA-PAINT single-particle tracking (DNA-PAINT-SPT) enables extended single-molecule studies of membrane protein interactions

Niederauer, C.; Nguyen, C.; Wang-Henderson, M.; Stein, J.; Strauss, S.; Cumberworth, A.; Stehr, F.; Jungmann, R.; Schwille, P.; Ganzinger, K. A.

doi:10.1101/2022.08.25.503948

bioRxiv · 10.1101/2022.08.25.503948

DNA-PAINT single-particle tracking (DNA-PAINT-SPT) enables extended single-molecule studies of membrane protein interactions

Niederauer, C.·Nguyen, C.·Wang-Henderson, M.·Stein, J.·Strauss, S.·Cumberworth, A.·Stehr, F.·Jungmann, R.·Schwille, P.·Ganzinger, K. A.

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Abstract

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-colour DNA-PAINT-SPT for quantifying protein dimerisation and tracking proteins in living cell membranes, demonstrating its improved performance over single-dye SPT.

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Niederauer, C., Nguyen, C., Wang-Henderson, M., Stein, J., Strauss, S., Cumberworth, A., Stehr, F., Jungmann, R., Schwille, P., Ganzinger, K. A.. 2022-08-25. DNA-PAINT single-particle tracking (DNA-PAINT-SPT) enables extended single-molecule studies of membrane protein interactions. https://doi.org/10.1101/2022.08.25.503948

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