Localized functional chemical stimulation of TE 671 cells cultured on nanoporous membrane by calcein and acetylcholine

Biophys J. 2007 Jan 1;92(1):L04-6. doi: 10.1529/biophysj.106.096743. Epub 2006 Nov 3.

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.

Publication types

  • Letter
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcholine / chemistry*
  • Benzimidazoles / pharmacology
  • Calcium / chemistry
  • Calcium / metabolism
  • Cell Adhesion
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods*
  • Eosine Yellowish-(YS) / chemistry
  • Fluoresceins / chemistry*
  • Fluoresceins / pharmacology
  • Fluorescent Dyes / pharmacology
  • Humans
  • Kinetics
  • Membranes, Artificial
  • Microscopy, Fluorescence
  • Models, Chemical
  • Tumor Cells, Cultured*

Substances

  • Benzimidazoles
  • Fluoresceins
  • Fluorescent Dyes
  • Membranes, Artificial
  • calcein AM
  • Acetylcholine
  • bisbenzimide ethoxide trihydrochloride
  • Calcium
  • Eosine Yellowish-(YS)
  • fluorexon