TY  - JOUR
A1  - Lukeš, Tomáš
A1  - Glatzová, Daniela
A1  - Kvíčalová, Zuzana
A1  - Levet, Florian
A1  - Benda, Aleš
A1  - Letschert, Sebastian
A1  - Sauer, Markus
A1  - Brdička, Tomáš
A1  - Lasser, Theo
A1  - Cebecauer, Marek
T1  - Quantifying protein densities on cell membranes using super-resolution optical fluctuation imaging
JF  - Nature Communications
N2  - 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.
KW  - biology
KW  - fluorescence imaging
KW  - imaging the immune system
KW  - super-resolution microscopy
Y1  - 2017
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-172993
VL  - 8
ER  -