title: Dynamics of intracellular processes in live-cell systems unveiled by fluorescence correlation microscopy.
abstract: Fluorescence fluctuation-based methods are non-invasive microscopy tools especially suited for the study of dynamical aspects of biological processes. These methods examine spontaneous intensity fluctuations produced by fluorescent molecules moving through the small, femtoliter-sized observation volume defined in confocal and multiphoton microscopes. The quantitative analysis of the intensity trace provides information on the processes producing the fluctuations that include diffusion, binding interactions, chemical reactions and photophysical phenomena. In this review, we present the basic principles of the most widespread fluctuation-based methods, discuss their implementation in standard confocal microscopes and briefly revise some examples of their applications to address relevant questions in living cells. The ultimate goal of these methods in the Cell Biology field is to observe biomolecules as they move, interact with targets and perform their biological action in the natural context. © 2016 IUBMB Life, 69(1):8-15, 2017.
authors.affiliation: Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina, IQUIBICEN, UBA-CONICET.
authors.forename: Nicolás
authors.initials: N
authors.lastname: González Bardeci
authors.affiliation: Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina, IQUIBICEN, UBA-CONICET.
authors.forename: Juan Francisco
authors.initials: JF
authors.lastname: Angiolini
authors.affiliation: Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina, IQUIBICEN, UBA-CONICET.
authors.forename: María Cecilia
authors.initials: MC
authors.lastname: De Rossi
authors.affiliation: IFIBA, UBA-CONICET, Argentina.
authors.forename: Luciana
authors.initials: L
authors.lastname: Bruno
authors.affiliation: Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina, IQUIBICEN, UBA-CONICET.
authors.forename: Valeria
authors.initials: V
authors.lastname: Levi
mesh_terms: D001692:Biological Transport; D002477:Cells / Q000648:ultrastructure*; D003593:Cytoplasm / Q000648:ultrastructure*; D005453:Fluorescence; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000379:methods*
keywords: autocorrelation; cross-correlation; diffusion; fluorescence correlation spectroscopy; fluorescence microscopy; intracellular dynamics