Article abstract


Nature Methods 6, 905 - 909 (2009)
Published online: 15 November 2009 | doi:10.1038/nmeth.1400

Cell stimulation with optically manipulated microsources

Holger Kress1, Jin-Gyu Park1, Cecile O Mejean1, Jason D Forster1, Jason Park2, Spencer S Walse3,8, Yong Zhang4, Dianqing Wu4, Orion D Weiner5, Tarek M Fahmy2,3 & Eric R Dufresne1,3,6,7


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 mum 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.

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  1. Department of Mechanical Engineering, Yale University, New Haven, Connecticut, USA.
  2. Department of Biomedical Engineering, Yale University, New Haven, Connecticut, USA.
  3. Department of Chemical Engineering, Yale University, New Haven, Connecticut, USA.
  4. Vascular Biology and Therapeutics Program and Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut, USA.
  5. Department of Biochemistry, University of California, San Francisco, San Francisco, California, USA.
  6. Department of Physics, Yale University New Haven, Connecticut, USA.
  7. Department of Cell Biology, Yale University New Haven, Connecticut, USA.
  8. Present address: US Department of Agriculture, Parlier, California, USA.

Correspondence to: Holger Kress1 e-mail: holger.kress@yale.edu

Correspondence to: Eric R Dufresne1,3,6,7 e-mail: eric.dufresne@yale.edu



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