Dynamics of intracellular processes in live-cell systems unveiled by fluorescence correlation microscopy

NICOLÁS GONZÁLEZ BARDECI, JUAN FRANCISCO ANGIOLINI, MARÍA CECILIA DE ROSSI, LUCIANA BRUNO AND VALERIA LEVI

IUBMB LIFE

JOHN WILEY & SONS INC

Año: 2017 vol. 69 p. 8 - 15

1521-6543

Fluorescence fluctuation-based methods are non-invasive microscopy toolsespecially suited for the study of dynamical aspects of biological processes. Thesemethods examine spontaneous intensity fluctuations produced by fluorescentmolecules moving through the small, femtoliter-sized observation volume definedin confocal and multiphoton microscopes. The quantitative analysis of theintensity trace provides information on the processes producing thefluctuations that include diffusion, binding interactions, chemical reactionsand photophysical phenomena. In this review, we present the basic principles ofthe most widespread fluctuation-based methods, discuss their implementation instandard confocal microscopes and briefly revise some examples of theirapplications to address relevant questions in living cells. The ultimate goalof these methods in the Cell Biology field is to observe biomolecules as theymove, interact with targets and perform their biological action in the naturalcontext.