title: Method for co-cluster analysis in multichannel single-molecule localisation data.
abstract: We demonstrate a combined univariate and bivariate Getis and Franklin's local point pattern analysis method to investigate the co-clustering of membrane proteins in two-dimensional single-molecule localisation data. This method assesses the degree of clustering of each molecule relative to its own species and relative to a second species. Using simulated data, we show that this approach can quantify the degree of cluster overlap in multichannel point patterns. The method is validated using photo-activated localisation microscopy and direct stochastic optical reconstruction microscopy data of the proteins Lck and CD45 at the T cell immunological synapse. Analysing co-clustering in this manner is generalizable to higher numbers of fluorescent species and to three-dimensional or live cell data sets.
authors.affiliation: Centre for Vascular Research and Australian Centre for NanoMedicine, University of New South Wales, Sydney, Australia.
authors.forename: Jérémie
authors.initials: J
authors.lastname: Rossy
authors.forename: Edward
authors.initials: E
authors.lastname: Cohen
authors.forename: Katharina
authors.initials: K
authors.lastname: Gaus
authors.forename: Dylan M
authors.initials: DM
authors.lastname: Owen
mesh_terms: D006801:Humans; D007091:Image Processing, Computer-Assisted; D054992:Immunological Synapses / Q000378:metabolism*; D019169:Jurkat Cells; D017493:Leukocyte Common Antigens / Q000032:analysis / Q000378:metabolism; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000032:analysis / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism
chemical_list: D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D017493:Leukocyte Common Antigens