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Dynamics of intracellular processes in live‐cell systems unveiled by fluorescence correlation microscopy
Author(s) -
González Bardeci Nicolás,
Angiolini Juan Francisco,
De Rossi María Cecilia,
Bruno Luciana,
Levi Valeria
Publication year - 2017
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1002/iub.1589
Subject(s) - biomolecule , context (archaeology) , confocal , biological system , microscope , fluorescence microscope , confocal microscopy , living cell , fluorescence correlation spectroscopy , microscopy , fluorescence , nanotechnology , biophysics , live cell imaging , dynamics (music) , chemistry , physics , biology , materials science , optics , cell , biochemistry , paleontology , acoustics
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