Localized functional chemical stimulation of TE 671 cells cultured on nanoporous membrane by calcein and acetylcholine
Susanne Zibek 1, Alfred Stett 1, Peter Koltay 2, Min Hu 2, Roland Zengerle 2, Wilfried Nisch 1 and Martin Stelzle 3*
1 Natural and Medical Sciences Institute
2 Institute of Microtechnology, University Freiburg, Germany
3 Naturwissenschaftliches und Medizinisches Institut
* To whom correspondence should be addressed. E-mail: stelzle{at}nmi.de.
Submitted on September 5, 2006
Revised on September 21, 2006
Accepted on 12 October 2006
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Abstract |
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Acetylcholine sensitive TE 671 cells were cultured on nanoporous membranes and chemically stimulated by localized application of i) calcein-AM and ii) acetylcholine, respectively, onto the bottom face of the membrane employing an ink jet print head. Stimulus correlated response of cells was recorded by fluorescence microscopy with temporal and spatial resolution. Calcein fluorescence develops as a result of intracellular enzymatic conversion of calcein-AM whereas Ca2+ imaging using Fluo4 dye was employed to visualize cellular response to acetylcholine stimulation. Using 25 pl droplets and substance concentration ranging from 10 µM to 1 mM on nucleoporeTM membranes with pore diameters between 50 nm and 1 µm, a resolution on the order of 50 µm was achieved.
Key Words:
Ca-imaging, acetylcholine, chemical stimulation, nanoporous membrane
