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Issue 18, 2010
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Microfluidic perfusion system for culturing and imaging yeast cell microarrays and rapidly exchanging media

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Abstract

High resolution live cell microscopy is increasingly used to detect cellular dynamics in response to drugs and chemicals, but it depends on complex and expensive liquid handling devices that have limited its wider adoption. Here, we present a microfluidic perfusion system that is built without using specialized microfabrication infrastructure, simple to use because only a pipette is needed for liquid handling, and yet allows for rapid media exchange and simultaneous fluorescence microscopy imaging. Yeast cells may be introduced from a culture, or spotted as arrays on a coverslip, and are sandwiched with a 20 μm thick track-etched membrane. A second coverslip and a mesh with 120 μm porosity are placed on top, forming a microfluidic conduit for lateral flow of solutions by capillary effects. Solutions introduced through the inlet flow through the mesh and chemicals diffuse vertically across the membrane to the cells trapped below. Solutions are exchanged by adding a new sample to the inlet. Using this system, we studied the dynamic response of F-actin in living yeast expressing Sac6-EGFP—a protein associated with discrete F-actin structures called “patches”—to the drug latrunculin A, a well known inhibitor of actin polymerization. We observed that the patches disappeared in 85% of the cells within 5 min, and re-assembled in 45 min following exchange of the drug with media. The perfusion system presented here is a simple, inexpensive device suited for analysis of drug dose-response and regeneration of single cells and arrays of cells.

Graphical abstract: Microfluidic perfusion system for culturing and imaging yeast cell microarrays and rapidly exchanging media

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Publication details

The article was received on 07 Apr 2010, accepted on 28 Jul 2010 and first published on 12 Aug 2010


Article type: Paper
DOI: 10.1039/C004857G
Lab Chip, 2010,10, 2449-2457

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    Microfluidic perfusion system for culturing and imaging yeast cell microarrays and rapidly exchanging media

    M. Mirzaei, M. Pla-Roca, R. Safavieh, E. Nazarova, M. Safavieh, H. Li, J. Vogel and D. Juncker, Lab Chip, 2010, 10, 2449
    DOI: 10.1039/C004857G

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