Dresden 2020 – wissenschaftliches Programm
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BP 35.1: Vortrag
Donnerstag, 19. März 2020, 15:00–15:15, HÜL 386
Molecule counts in complex oligomers with single-molecule localization microscopy — Tim Niklas Baldering1, •Jakob Tómas Bullerjahn2, Gerhard Hummer2, Mike Heilemann1, and Sebastian Malkusch1 — 1Institute of Physical and Theoretical Chemistry, Goethe-University Frankfurt, Frankfurt am Main, Germany — 2Department of Theoretical Biophysics, Max Planck Institute of Biophysics, Frankfurt am Main, Germany
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 photoactivated localization microscopy (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.