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Cellular Cartography…Mapping Protein Dynamics in Living Cells with novel Image Correlation Techniques
Author(s) -
Wiseman Paul W.
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.12.3
Subject(s) - digital image correlation , integrin , fluorescence correlation spectroscopy , biological system , microbiology and biotechnology , biophysics , extracellular matrix , image (mathematics) , chemistry , biology , physics , computer science , cell , artificial intelligence , biochemistry , fluorescence , quantum mechanics , optics
Image correlation methods provide a new window of analysis for measurement of protein‐protein interactions and macromolecular transport properties from fluorescence images of living cells. We recently introduced spatio‐temporal image correlation spectroscopy (STICS) which measures vectors of protein flux in cells based on the calculation of a spatial correlation function as a function of time from an image time series. Here we will describe the application of STICS for measuring transport maps of adhesion related macromolecules such as integrin, alpha‐actinin, paxillin, talin, etc. within, or associated with the basal membrane in living fibroblast and CHO cells. These measurements have allowed us to propose a model for the molecular clutch that regulates connections between the extracellular matrix, integrins in the membrane and the cytoskeleton during cell protrusion and migration. We will also highlight recent advances we have made with a new form of reciprocal space ICS, called kICS, that allows us to measure unbiased transport coefficients and clustering of fluorescently labeled membrane proteins even if there is complex photophysics (such as nanoparticle emission blinking) of the probe.