title: Simultaneous multi-species tracking in live cells with quantum dot conjugates.
abstract: Quantum dots are available in a range of spectrally separated emission colors and with a range of water-stabilizing surface coatings that offers great flexibility for enabling bio-specificity. In this study, we have taken advantage of this flexibility to demonstrate that it is possible to perform a simultaneous investigation of the lateral dynamics in the plasma membrane of i) the transmembrane epidermal growth factor receptor, ii) the glucosylphospatidylinositol-anchored protein CD59, and iii) ganglioside GM1-cholera toxin subunit B clusters in a single cell. We show that a large number of the trajectories are longer than 50 steps, which we by simulations show to be sufficient for robust single trajectory analysis. This analysis shows that the populations of the diffusion coefficients are heterogeneously distributed for all three species, but differ between the different species. We further show that the heterogeneity is decreased upon treating the cells with methyl-β-cyclodextrin.
authors.affiliation: MEMPHYS - Center for Biomembrane Physics and Danish Molecular Biomedical Imaging Center (DaMBIC), University of Southern Denmark, Odense M, Denmark.
authors.forename: Mathias P
authors.initials: MP
authors.lastname: Clausen
authors.affiliation: MEMPHYS - Center for Biomembrane Physics and Danish Molecular Biomedical Imaging Center (DaMBIC), University of Southern Denmark, Odense M, Denmark.
authors.forename: Eva C
authors.initials: EC
authors.lastname: Arnspang
authors.affiliation: Molecular Biosensor and Imaging Center (MBIC), Mellon Institute, Carnegie Mellon University, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Byron
authors.initials: B
authors.lastname: Ballou
authors.affiliation: Lineberger Comprehensive Cancer Center and Department of Cell and Developmental Biology, Howard Hughes Medical Institute, University of North Carolina, Chapel Hill, North Carolina, United States of America.
authors.forename: James E
authors.initials: JE
authors.lastname: Bear
authors.affiliation: MEMPHYS - Center for Biomembrane Physics and Danish Molecular Biomedical Imaging Center (DaMBIC), University of Southern Denmark, Odense M, Denmark.
authors.forename: B Christoffer
authors.initials: BC
authors.lastname: Lagerholm
mesh_terms: D000818:Animals; D018957:CD59 Antigens / Q000032:analysis*; D002470:Cell Survival; D058948:Cell Tracking / Q000379:methods*; D003198:Computer Simulation; D004058:Diffusion; D066246:ErbB Receptors / Q000032:analysis*; D005677:G(M1) Ganglioside / Q000032:analysis*; D021621:Imaging, Three-Dimensional; D008623:Mercaptoethanol / Q000494:pharmacology; D051379:Mice; D009010:Monte Carlo Method; D045663:Quantum Dots / Q000378:metabolism*; D015203:Reproducibility of Results; D013194:Staining and Labeling; D019809:Streptavidin / Q000378:metabolism
chemical_list: D018957:CD59 Antigens; D005677:G(M1) Ganglioside; D008623:Mercaptoethanol; D019809:Streptavidin; D066246:ErbB Receptors