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  Molecule counts in complex oligomers with single-molecule localization microscopy

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.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0004-C094-B Version Permalink: http://hdl.handle.net/21.11116/0000-0004-C095-A
Genre: Journal Article

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 Creators:
Baldering, Tim N.1, Author
Bullerjahn, Jakob T.2, Author              
Hummer, Gerhard2, Author              
Heilemann, Mike1, Author
Malkusch, Sebastian1, Author
Affiliations:
1Institute of Physical and Theoretical Chemistry, Goethe-University Frankfurt, Frankfurt am Main, Germany, ou_persistent22              
2Department of Theoretical Biophysics, Max Planck Institute of Biophysics, Max Planck Society, ou_2068292              

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Free keywords: fluorescent proteins, molecular counting, photoactivated-localization microscopy, single-molecule localization microscopy
 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.

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Language(s): eng - English
 Dates: 2019-06-082019-08-152019-09-062019-11-20
 Publication Status: Published in print
 Pages: 14
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1088/1361-6463/ab3b65
BibTex Citekey: baldering_molecule_2019
 Degree: -

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Title: Journal of Physics D: Applied Physics
  Abbreviation : J. Phys. D: Appl. Phys.
Source Genre: Journal
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Publ. Info: Bristol : IOP Publishing
Pages: - Volume / Issue: 52 (47) Sequence Number: 474002 Start / End Page: - Identifier: ISSN: 0022-3727
CoNE: https://pure.mpg.de/cone/journals/resource/0022-3727