A microfluidic device for reversible environmental changes around single cells using optical tweezers for cell selection and positioning
Artikel i vetenskaplig tidskrift, 2010

Cells naturally exist in a dynamic chemical environment, and therefore it is necessary to study cell behaviour under dynamic stimulation conditions in order to understand the signalling transduction pathways regulating the cellular response. However, until recently, experiments looking at the cellular response to chemical stimuli have mainly been performed by adding a stress substance to a population of cells and thus only varying the magnitude of the stress. In this paper we demonstrate an experimental method enabling acquisition of data on the behaviour of single cells upon reversible environmental perturbations, where microfluidics is combined with optical tweezers and fluorescence microscopy. The cells are individually selected and positioned in the measurement region on the bottom surface of the microfluidic device using optical tweezers. The optical tweezers thus enable precise control of the cell density as well as the total number of cells within the measurement region. Consequently, the number of cells in each experiment can be optimized while clusters of cells, that render subsequent image analysis more difficult, can be avoided. The microfluidic device is modelled and demonstrated to enable reliable changes between two different media in less than 2 s. The experimental method is tested by following the cycling of GFP-tagged proteins (Mig1 and Msn2, respectively) between the cytosol and the nucleus in Saccharomyces cerevisiae upon changes in glucose availability.

Författare

Emma Eriksson

Göteborgs universitet

Kristin Sott

SuMo Biomaterials

Göteborgs universitet

Fredrik Lundqvist

Göteborgs universitet

Martin Sveningsson

Göteborgs universitet

Jan Scrimgeour

Dag Hanstorp

Göteborgs universitet

Mattias Goksör

Göteborgs universitet

Annette Graneli

Göteborgs universitet

Lab on a Chip - Miniaturisation for Chemistry and Biology

1473-0197 (ISSN) 1473-0189 (eISSN)

Vol. 10 617-625

Ämneskategorier

Annan fysik

Den kondenserade materiens fysik

DOI

10.1039/b913587a