Advances in image correlation spectroscopy: measuring number densities, aggregation states, and dynamics of fluorescently labeled macromolecules in cells.
A brief historical outline of fluorescence fluctuation correlation techniques is presented, followed by an in-depth review of the theory and development of image correlation techniques, including: image correlation spectroscopy (ICS), temporal ICS (TICS), image cross-correlation spectroscopy (ICCS), spatiotemporal ICS (STICS), k-space ICS (kICS), raster ICS (RICS), and particle ICS (PICS). These techniques can be applied to analyze image series acquired on commercially available laser scanning or total internal reflection fluorescence microscopes, and are used to determine the number density, aggregation state, diffusion coefficient, velocity, and interaction fraction of fluorescently labeled molecules or particles. A comprehensive review of the application of ICS techniques to a number of systems, including cell adhesion, membrane receptor aggregation and dynamics, virus particle fusion, and fluorophore photophysics, is presented.
Kolin
David L
DL
Department of Chemistry, McGill University, Montreal, QC, Canada H3A 2K6.
Wiseman
Paul W
PW
D000185:Actinin / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D007091:Image Processing, Computer-Assisted; D051379:Mice; D008856:Microscopy, Fluorescence; D051419:Paxillin / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013057:Spectrum Analysis / Q000379:methods*; D013997:Time Factors; D014162:Transfection
D005456:Fluorescent Dyes; D051419:Paxillin; D011993:Recombinant Fusion Proteins; D000185:Actinin; D049452:Green Fluorescent Proteins