Steady-state cross-correlations for live two-colour super-resolution localization data sets

Cross-correlation of super-resolution images gathered from point localizations allows for robust quantification of protein co-distributions in chemically fixed cells. Here this is extended to dynamic systems through an analysis that quantifies the steady-state cross-correlation between spectrally distinguishable probes. This methodology is used to quantify the co-distribution of several mobile membrane proteins in both vesicles and live cells, including Lyn kinase and the B-cell receptor during antigen stimulation.

[1]  C. Bustamante,et al.  Counting single photoactivatable fluorescent molecules by photoactivated localization microscopy (PALM) , 2012, Proceedings of the National Academy of Sciences.

[2]  S. Semrau,et al.  Quantification of biological interactions with particle image cross-correlation spectroscopy (PICCS). , 2011, Biophysical journal.

[3]  J. Cambier,et al.  Phosphorylated immunoreceptor signaling motifs (ITAMs) exhibit unique abilities to bind and activate Lyn and Syk tyrosine kinases. , 1995, Journal of immunology.

[4]  Watt W. Webb,et al.  Temporally resolved interactions between antigen-stimulated IgE receptors and Lyn kinase on living cells , 2005, The Journal of cell biology.

[5]  J. D. Dal Porto,et al.  B cell antigen receptor signaling 101. , 2004, Molecular immunology.

[6]  Benjamin B. Machta,et al.  Correlation Functions Quantify Super-Resolution Images and Estimate Apparent Clustering Due to Over-Counting , 2011, PloS one.

[7]  Santiago Costantino,et al.  A guide to accurate fluorescence microscopy colocalization measurements. , 2006, Biophysical journal.

[8]  S. Pierce,et al.  The tipping points in the initiation of B cell signalling: how small changes make big differences , 2010, Nature Reviews Immunology.

[9]  M. Heilemann,et al.  Carbocyanine dyes as efficient reversible single-molecule optical switch. , 2005, Journal of the American Chemical Society.

[10]  P. Schwille,et al.  Partitioning, diffusion, and ligand binding of raft lipid analogs in model and cellular plasma membranes. , 2012, Biochimica et biophysica acta.

[11]  Benjamin B Machta,et al.  Liquid general anesthetics lower critical temperatures in plasma membrane vesicles. , 2013, Biophysical journal.

[12]  A. Burkhardt,et al.  Anti-immunoglobulin stimulation of B lymphocytes activates src-related protein-tyrosine kinases. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Michael J Rust,et al.  Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM) , 2006, Nature Methods.

[14]  P. Sengupta,et al.  Quantitative nanoscale analysis of IgE-FcεRI clustering and coupling to early signaling proteins. , 2012, The journal of physical chemistry. B.

[15]  J. Lippincott-Schwartz,et al.  Imaging Intracellular Fluorescent Proteins at Nanometer Resolution , 2006, Science.

[16]  P. Annibale,et al.  Quantitative Photo Activated Localization Microscopy: Unraveling the Effects of Photoblinking , 2011, PloS one.

[17]  J. Spudich,et al.  Single molecule high-resolution colocalization of Cy3 and Cy5 attached to macromolecules measures intramolecular distances through time. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Michael D. Mason,et al.  Ultra-high resolution imaging by fluorescence photoactivation localization microscopy. , 2006, Biophysical journal.

[19]  S. Semrau,et al.  Particle image correlation spectroscopy (PICS): retrieving nanometer-scale correlations from high-density single-molecule position data. , 2007, Biophysical journal.

[20]  W E Moerner,et al.  Quantitative multicolor subdiffraction imaging of bacterial protein ultrastructures in three dimensions. , 2013, Nano letters.

[21]  B. Sefton,et al.  Protein tyrosine phosphorylation is induced in murine B lymphocytes in response to stimulation with anti‐immunoglobulin. , 1990, The EMBO journal.

[22]  X. Zhuang,et al.  Evaluation of Fluorophores for Optimal Performance in Localization-Based Super-Resolution Imaging , 2012 .

[23]  Yongdeng Zhang,et al.  Rational design of true monomeric and bright photoactivatable fluorescent proteins , 2012, Nature Methods.

[24]  Enrico Gratton,et al.  Fast Spatiotemporal Correlation Spectroscopy to Determine Protein Lateral Diffusion Laws in Live Cell Membranes , 2014 .

[25]  Carla Coltharp,et al.  Accurate Construction of Photoactivated Localization Microscopy (PALM) Images for Quantitative Measurements , 2012, PloS one.

[26]  Prabuddha Sengupta,et al.  Probing protein heterogeneity in the plasma membrane using PALM and pair correlation analysis , 2011, Nature Methods.

[27]  Robert G. Parton,et al.  Direct visualization of Ras proteins in spatially distinct cell surface microdomains , 2003, The Journal of cell biology.

[28]  B. Baird,et al.  Cross-correlation analysis of inner-leaflet-anchored green fluorescent protein co-redistributed with IgE receptors and outer leaflet lipid raft components. , 2001, Biophysical journal.

[29]  X. Zhuang,et al.  Fast three-dimensional super-resolution imaging of live cells , 2011, Nature Methods.

[30]  B. Baird,et al.  Distinct stages of stimulated FcεRI receptor clustering and immobilization are identified through superresolution imaging. , 2013, Biophysical journal.

[31]  Tobias M. P. Hartwich,et al.  Video-rate nanoscopy using sCMOS camera- specific single-molecule localization algorithms , 2013 .

[32]  Suliana Manley,et al.  Functional nanoscale organization of signaling molecules downstream of the T cell antigen receptor. , 2011, Immunity.

[33]  A. Clayton,et al.  Fixation alters fluorescence lifetime and anisotropy of cells expressing EYFP-tagged serotonin1A receptor. , 2011, Biochemical and biophysical research communications.

[34]  Steve Pressé,et al.  Stochastic approach to the molecular counting problem in superresolution microscopy , 2014, Proceedings of the National Academy of Sciences.