Characterization of a New Series of Fluorescent Probes for Imaging Membrane Order |
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| Authors: | Joanna M. Kwiatek Dylan M. Owen Ahmed Abu-Siniyeh Ping Yan Leslie M. Loew Katharina Gaus |
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| Affiliation: | 1. Centre for Vascular Research and Australian Centre for Nanomedicine, University of New South Wales, Sydney, Australia.; 2. Center for Cell Analysis and Modelling, University of Connecticut Health Center, Farmington, Connecticut, United States of America.; Institut Curie, France, |
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| Abstract: | Visualization and quantification of lipid order is an important tool in membrane biophysics and cell biology, but the availability of environmentally sensitive fluorescent membrane probes is limited. Here, we present the characterization of the novel fluorescent dyes PY3304, PY3174 and PY3184, whose fluorescence properties are sensitive to membrane lipid order. In artificial bilayers, the fluorescence emission spectra are red-shifted between the liquid-ordered and liquid-disordered phases. Using ratiometric imaging we demonstrate that the degree of membrane order can be quantitatively determined in artificial liposomes as well as live cells and intact, live zebrafish embryos. Finally, we show that the fluorescence lifetime of the dyes is also dependent on bilayer order. These probes expand the current palate of lipid order-sensing fluorophores affording greater flexibility in the excitation/emission wavelengths and possibly new opportunities in membrane biology. |
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