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The lateral organization of molecules in the cellular plasma membrane plays an important role in cellular signaling. A critical parameter for membrane molecular organization is how the membrane lipids are packed. Polarity-sensitive dyes are powerful tools to characterize such lipid membrane order, employing, for example, confocal and two-photon microscopy. The investigation of potential nanodomains, however, requires the use of superresolution microscopy. Here, we test the performance of the polarity-sensitive membrane dyes Di-4-ANEPPDHQ, Di-4-AN(F)EPPTEA, and NR12S in superresolution stimulated emission depletion microscopy. Measurements on cell-derived membrane vesicles, in the plasma membrane of live cells, and on single virus particles, show the high potential of these dyes for probing nanoscale membrane heterogeneity.

Original publication




Journal article


Biophys J

Publication Date





1321 - 1330


Animals, Benzoxazines, CHO Cells, Cell Membrane, Cricetulus, Cytoplasmic Vesicles, Fluorescent Dyes, HEK293 Cells, HIV, Humans, Microscopy, Molecular Structure, Phosphatidylethanolamines, Pyridinium Compounds, Quaternary Ammonium Compounds, Spectrum Analysis, Virion, Voltage-Sensitive Dye Imaging