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About the Authors:
Joanna M. Kwiatek
Contributed equally to this work with: Joanna M. Kwiatek, Dylan M. Owen
Affiliation: Centre for Vascular Research and Australian Centre for Nanomedicine, University of New South Wales, Sydney, Australia
Dylan M. Owen
Contributed equally to this work with: Joanna M. Kwiatek, Dylan M. Owen
* E-mail: [email protected] (DMO), [email protected] (LML), [email protected] (KG)
Affiliation: Centre for Vascular Research and Australian Centre for Nanomedicine, University of New South Wales, Sydney, Australia
Ahmed Abu-Siniyeh
Affiliation: Centre for Vascular Research and Australian Centre for Nanomedicine, University of New South Wales, Sydney, Australia
Ping Yan
Affiliation: Center for Cell Analysis and Modelling, University of Connecticut Health Center, Farmington, Connecticut, United States of America
Leslie M. Loew
* E-mail: [email protected] (DMO), [email protected] (LML), [email protected] (KG)
Affiliation: Center for Cell Analysis and Modelling, University of Connecticut Health Center, Farmington, Connecticut, United States of America
Katharina Gaus
* E-mail: [email protected] (DMO), [email protected] (LML), [email protected] (KG)
Affiliation: Centre for Vascular Research and Australian Centre for Nanomedicine, University of New South Wales, Sydney, Australia
Introduction
Lipids and proteins are not homogenously distributed in the plasma membrane of eukaryotic cells giving raise to biochemically and biophysically distinct domains. In artificial bilayers, the membrane can separate into immiscible liquid-ordered and liquid-disordered phases that differ in the degree of lipid packing. Highly ordered domains, known as lipid rafts, have been postulated to also exist in the cell membrane with implications for protein distributions and diffusion [1], [2], [3], [4]. Numerous cellular processes including receptor signaling, for example at the immunological synapse of T cells [5], [6], and membrane trafficking [7], for example in polarized epithelial cells 8,9] are thought to be influenced by membrane order. Lipid domains with a high membrane order have also been shown to play a role in a number of pathologies [10], [11].
Fluorescence microscopy is the tool-of-choice to investigate membrane lipid domains [12], [13] as it can equally be applied to model and cell membranes and avoids fixation, which can introduce artifacts in lipid and protein organization [14]. The properties of lipid domains can be investigated by analyzing the localization and dynamics of membrane proteins and lipids [15], however, most of these approaches rely on prior knowledge of the preference of the probe for partitioning into the...