Abstract
Quantitative measurement of diffusive and directional processes of intracellular structures is not only critical in understanding cell mechanics and functions, but also has many applications, such as investigation of cellular responses to therapeutic agents. We introduce a label-free optical technique that allows non-perturbative characterization of localized intracellular dynamics. The method combines a field-based dynamic light scattering analysis with a confocal interferometric microscope to provide a statistical measure of the diffusive and directional motion of scattering structures inside a microscopic probe volume. To demonstrate the potential of this technique, we examined the localized intracellular dynamics in human epithelial ovarian cancer cells. We observed the distinctive temporal regimes of intracellular dynamics, which transitions from random to directional processes on a timescale of ~0.01 sec. In addition, we observed disrupted directional processes on the timescale of 1~5 sec by the application of a microtubule polymerization inhibitor, Colchicine, and ATP depletion.
Highlights
Cells are highly dynamic systems that continuously undergo internal reconfiguration through random and/or coordinated molecular and mechanical responses [1]
We show the validity of F-Dynamic light scattering (DLS) analysis based on the measurement of emulsion particles, and demonstrate field-based dynamic light scattering (F-DLS) as a tool for characterizing localized intracellular dynamics inside human ovarian cancer cells (OVCAR-5s)
This study demonstrates the potential of the F-DLS technique in characterizing mechanical properties of dynamic systems, which could impact a broad range of biomedical applications
Summary
Cells are highly dynamic systems that continuously undergo internal reconfiguration through random and/or coordinated molecular and mechanical responses [1]. Intracellular dynamics such as the transport of molecules and cargos, along with cytoskeleton rearrangements, are fundamental processes that support a broad range of functions such as cell migration and division. These intracellular processes are derived from, and often influence physiological conditions of the cells. Quantitative measurement of intracellular processes would aid in building a better understanding of the underlying mechanisms of cellular states and functions
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