Electrically evoking and electrochemically resolving quantal release on a microchip.
A microchip was applied to electrically depolarize rat pheochromocytoma (PC12) cells and to simultaneously detect exocytotic catecholamine release amperometrically. Results demonstrate exocytosis elicited by flowing cells through an electric field generated by a potentiostat circuit in a microchannel, as well as exocytosis triggered by application of an extracellular voltage pulse across. Electrical finite element model (FEM) analysis illustrated that larger cells experienced greater depolarizing excitation from the extracellular electric fields due to the smaller shunt path and higher resistance to current flow in the channel around the cell. Consistent with these simulations, data recorded from cell clusters and large cells exhibited increased release rates relative to data from the smaller cells. Overall, the system was capable of resolving single vesicle quantal release, in the zeptomole range, as well as the kinetics associated with the vesicle fusion process. Analysis of spike population statistics suggested detection of catecholamines from multiple release sites around the cells. The potential for such a device to be used in flow cytometry to evoke and detect exocytosis was demonstrated.
Dittami
Gregory M
GM
Department of Bioengineering, University of Utah, Salt Lake City, UT 84118, USA. dittami@eng.utah.edu
Rabbitt
Richard D
RD
D000818:Animals; D002395:Catecholamines / Q000032:analysis*; D018929:Cell Culture Techniques; D048429:Cell Size; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D005089:Exocytosis; D056656:Lab-On-A-Chip Devices; D008564:Membrane Potentials / Q000502:physiology; D046208:Microchip Analytical Procedures / Q000379:methods*; D008839:Microelectrodes; D016716:PC12 Cells; D018408:Patch-Clamp Techniques; D051381:Rats
D002395:Catecholamines