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A novel nanocapillary electrophoretic electrochemical (Nano-CEEC) chip has been developed to demonstrate the possibility of zeptomole-level detection of neurotransmitters released from single living cells. The chip integrates three subunits to collect and concentrate scarce neurotransmitters released from single PC-12 cells, including a pair of targeting electrodes for single cells captured by controlling the surface charge density; a dual-asymmetry electrokinetic flow device for sample collection, pre-concentration and separation in a nanochannel; and an online electrochemical detector for zeptomole-level sample detection. This Nano-CEEC chip integrates a polydimethylsiloxane microchannel for cell sampling and biomolecule separation and a silicon dioxide nanochannel for sample pre-concentration and amperometric detection. The cell-capture voltage ranges from 0.1 to 1.5 V with a frequency of 1-10 kHz for PC-12 cells, and the single cell-capture efficiency is optimized by varying the duration of the applied field. All of the processes, from cell sampling to neurotransmitter detection, can be completed within 15 min. Catecholamines, including dopamine and norepinephrine (noradrenaline) released from coupled single cells, have been successfully detected using the Nano-CEEC chip. A detection limit of 30-75 zeptomoles was achieved, which is close to the levels released by a single neuron in vitro.

Nanocapillary electrophoretic electrochemical chip: towards analysis of biochemicals released by single cells