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Cell stimulation with optically manipulated microsources.

ABSTRACT: Molecular gradients are important for various biological processes including the polarization of tissues and cells during embryogenesis and chemotaxis. Investigations of these phenomena require control over the chemical microenvironment of cells. We present a technique to set up molecular concentration patterns that are chemically, spatially and temporally flexible. Our strategy uses optically manipulated microsources, which steadily release molecules. Our technique enables the control of molecular concentrations over length scales down to about 1 microm and timescales from fractions of a second to an hour. We demonstrate this technique by manipulating the motility of single human neutrophils. We induced directed cell polarization and migration with microsources loaded with the chemoattractant formyl-methionine-leucine-phenylalanine. Furthermore, we triggered highly localized retraction of lamellipodia and redirection of polarization and migration with microsources releasing cytochalasin D, an inhibitor of actin polymerization.

SUBMITTER: Kress H

PROVIDER: S-EPMC2813772 | biostudies-literature | 2009 Dec

REPOSITORIES: biostudies-literature

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Cell stimulation with optically manipulated microsources.

Kress Holger H Park Jin-Gyu JG Mejean Cecile O CO Forster Jason D JD Park Jason J Walse Spencer S SS Zhang Yong Y Wu Dianqing D Weiner Orion D OD Fahmy Tarek M TM Dufresne Eric R ER

Nature methods 20091115 12

Molecular gradients are important for various biological processes including the polarization of tissues and cells during embryogenesis and chemotaxis. Investigations of these phenomena require control over the chemical microenvironment of cells. We present a technique to set up molecular concentration patterns that are chemically, spatially and temporally flexible. Our strategy uses optically manipulated microsources, which steadily release molecules. Our technique enables the control of molecu ...[more]

PMID: 19915561

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Project description:Cell membrane capacitance and conductance are key pieces of intrinsic information correlated with the cellular dielectric parameters and morphology of the plasma membrane; these parameters have been used as electrophysiological biomarkers to characterize cellular phenotype and state, and they have many associated clinical applications. Here, we present our work on the non-invasive determination of cell membrane capacitance and conductance by an optically activated microfluidics chip. The model for determining the cell membrane capacitance and conductance was established by a single layer of the shell-core polarization model. Three-dimensional finite-element analyses of the positive and negative optically induced dielectrophoresis forces generated by the projected light arrays of spots were performed, thus providing a theoretical validation of the feasibility of this approach. Then, the crossover frequency spectra for four typical types of cells (Raji cells, MCF-7 cells, HEK293 cells, and K562 cells) were experimentally investigated by using a micro-vision based motion-tracking technique. The different responses of these cells to the positive and negative ODEP forces were studied under four different liquid conductivities by automatic observation and tracking of the cellular trajectory and texture during the cells' translation. The cell membrane capacitance and conductance were determined from the curve-fitted spectra, which were 11.1 ± 0.9 mF/m2 and 782 ± 32 S/m2, respectively, for Raji cells, 11.5 ± 0.8 mF/m2 and 114 ± 28 S/m2 for MCF-7 cells, 9.0 ± 0.9 mF/m2 and 187 ± 22 S/m2 for HEK293 cells, and 10.2 ± 0.7 mF/m2 and 879 ± 24 S/m2 for K562 cells. Furthermore, as an application of this technique, the membrane capacitances of MCF-7 cells treated with four different concentrations of drugs were acquired. This technique introduces a determination of cell membrane capacitance and conductance that yields statistically significant data while allowing information from individual cells to be obtained in a non-invasive manner.

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Cell stimulation with optically manipulated microsources.

Publications

Cell stimulation with optically manipulated microsources.

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