Integration of microdialysis sampling and microchip electrophoresis with electrochemical detection.
Here we describe the fabrication, optimization, and application of a microfluidic device that integrates microdialysis (MD) sampling, microchip electrophoresis (ME), and electrochemical detection (EC). The manner in which the chip is produced is reproducible and enables the fixed alignment of the MD/ME and ME/EC interfaces. Poly(dimethylsiloxane) (PDMS)-based valves were used for the discrete injection of sample from the hydrodynamic MD dialysate stream into a separation channel for analysis with ME. To enable the integration of ME with EC detection, a palladium decoupler was used to isolate the high voltages associated with electrophoresis from micrometer-sized carbon ink detection electrodes. Optimization of the ME/EC interface was needed to allow the use of biologically appropriate perfusate buffers containing high salt content. This optimization included changes in the fabrication procedure, increases in the decoupler surface area, and a programmed voltage shutoff. The ability of the MD/ME/EC system to sample a biological system was demonstrated by using a linear probe to monitor the stimulated release of dopamine from a confluent layer of PC 12 cells. To our knowledge, this is the first report of a microchip-based system that couples microdialysis sampling with microchip electrophoresis and electrochemical detection.
Mecker
Laura C
LC
Department of Chemistry, Saint Louis University, 3501 Laclede Avenue, St. Louis, Missouri 63103, USA.
Martin
R Scott
RS
D000311:Adrenal Glands / Q000166:cytology; D000818:Animals; D002460:Cell Line; D002477:Cells / Q000378:metabolism*; D004298:Dopamine / Q000032:analysis* / Q000378:metabolism*; D004563:Electrochemistry / Q000379:methods; D046209:Electrophoresis, Microchip / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D017551:Microdialysis / Q000295:instrumentation* / Q000379:methods; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D051381:Rats
D004298:Dopamine