Quantifying protein densities on cell membranes using super-resolution optical fluctuation imaging

0482642 - ÚFCH JH 2018 RIV GB eng J - Journal Article
Lukeš, T. - Glatzová, Daniela - Kvíčalová, Zuzana - Levet, F. - Benda, Aleš - Letschert, S. - Sauer, M. - Brdička, Tomáš - Lasser, T. - Cebecauer, Marek
Quantifying protein densities on cell membranes using super-resolution optical fluctuation imaging.
Nature Communications. Roč. 8, č. 1 (2017), č. článku 1731. E-ISSN 2041-1723
R&D Projects: GA ČR GA15-06989S
Institutional support: RVO:61388955 ; RVO:68378050
Keywords : quantifying protein densities * membranes * single-molecule localization microscopy
OECD category: Biochemistry and molecular biology
Impact factor: 12.353, year: 2017
Method of publishing: Open access

Quantitative approaches for characterizing molecular organization of cell membrane molecules under physiological and pathological conditions profit from recently developed super-resolution imaging techniques. Current tools employ statistical algorithms to determine clusters of molecules based on single-molecule localization microscopy (SMLM) data. These approaches are limited by the ability of SMLM techniques to identify and localize molecules in densely populated areas and experimental conditions of sample preparation and image acquisition. We have developed a robust, model-free, quantitative clustering analysis to determine the distribution of membrane molecules that excels in densely labeled areas and is tolerant to various experimental conditions, i.e. multiple-blinking or high blinking rates. The method is based on a TIRF microscope followed by a super-resolution optical fluctuation imaging (SOFI) analysis. The effectiveness and robustness of the method is validated using simulated and experimental data investigating nanoscale distribution of CD4 glycoprotein mutants in the plasma membrane of T cells.
Permanent Link: http://hdl.handle.net/11104/0278057