title: Localized functional chemical stimulation of TE 671 cells cultured on nanoporous membrane by calcein and acetylcholine.
abstract: 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 Ca(2+) imaging using fluo-4 dye was employed to visualize cellular response to acetylcholine stimulation. Using 25 pl droplets and substance concentration ranging from 10 microM to 1 mM on Nucleopore membranes with pore diameters between 50 nm and 1 microm, a resolution on the order of 50 microm was achieved.
authors.forename: Susanne
authors.initials: S
authors.lastname: Zibek
authors.forename: Alfred
authors.initials: A
authors.lastname: Stett
authors.forename: Peter
authors.initials: P
authors.lastname: Koltay
authors.forename: Min
authors.initials: M
authors.lastname: Hu
authors.forename: Roland
authors.initials: R
authors.lastname: Zengerle
authors.forename: Wilfried
authors.initials: W
authors.lastname: Nisch
authors.forename: Martin
authors.initials: M
authors.lastname: Stelzle
mesh_terms: D000109:Acetylcholine / Q000737:chemistry*; D001562:Benzimidazoles / Q000494:pharmacology; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D004801:Eosine Yellowish-(YS) / Q000737:chemistry; D005452:Fluoresceins / Q000737:chemistry* / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence; D008956:Models, Chemical; D014407:Tumor Cells, Cultured
chemical_list: D001562:Benzimidazoles; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008567:Membranes, Artificial; C085925:calcein AM; D000109:Acetylcholine; C017807:bisbenzimide ethoxide trihydrochloride; D002118:Calcium; D004801:Eosine Yellowish-(YS); C007740:fluorexon