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Molecule counts in complex oligomers with single-molecule localization microscopy

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Bullerjahn,  Jakob T.
Department of Theoretical Biophysics, Max Planck Institute of Biophysics, Max Planck Society;

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Hummer,  Gerhard
Department of Theoretical Biophysics, Max Planck Institute of Biophysics, Max Planck Society;

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Citation

Baldering, T. N., Bullerjahn, J. T., Hummer, G., Heilemann, M., & Malkusch, S. (2019). Molecule counts in complex oligomers with single-molecule localization microscopy. Journal of Physics D: Applied Physics, 52(47): 474002. doi:10.1088/1361-6463/ab3b65.


Cite as: http://hdl.handle.net/21.11116/0000-0004-C094-B
Abstract
Single-molecule localization microscopy resolves nano-scale protein clusters in cells, and in addition can extract protein copy numbers from within these clusters. A powerful approach for such molecular counting is the analysis of fluorophore blinking using stochastic model functions. Here, we develop a theoretical model for quantitative analysis of PALM data that accounts for the detection efficiency. By this, we are able to extract populations of different oligomers reliably and in complex mixtures. We demonstrate this approach analyzing simulated PALM data of a photoactivatable fluorescent protein. We generate simulations of blinking data of oligomers and of mixtures of oligomers, and show robust oligomer identification. In addition, we demonstrate this approach for experimental PALM data.