Continuous operation of microfabricated electrophoresis devices for 24 hours and application to chemical monitoring of living cells.
Microchip electrophoresis is an emerging analytical technology with several useful attributes including rapid separation time, small sample requirements, and automation. In numerous potential applications, such as chemical monitoring or high-throughput screening, it may be desirable to use a system for many analyses without operator intervention; however, long-term operation of microchip electrophoresis systems has received little attention. We have developed a microchip electrophoresis system that can automatically inject samples at 6 s intervals for 24 h resulting in collection of 14,400 assays in one session. Continuous operation time of a prototype of the device was limited to 2 h due to degradation of reagents and electrophoresis buffers on the chip; however, modification so that all reagents were continuously perfused into reservoirs on the device ensured fresh reagents were always used for analysis and enabled extended operating sessions. The electrophoresis chip incorporated a cell perfusion chamber and reagent addition channels to allow chemical monitoring of fluid around cells cultured on the chip by serial electrophoretic immunoassays. The immunoassay had detection limits of 0.4 nM for insulin and generated approximately 4% relative standard deviation over an entire 24 h period with no evidence of signal drift. The combined system was used to monitor insulin secretion from single islets of Langerhans for 6-39 h. The monitoring experiments revealed that islets have secretion dynamics that include spontaneous oscillations after extended nonoscillating periods and possible ultradian rhythms.
Reid
Kendra R
KR
Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.
Kennedy
Robert T
RT
D000818:Animals; D002478:Cells, Cultured; D046209:Electrophoresis, Microchip / Q000379:methods*; D007118:Immunoassay / Q000379:methods; D007202:Indicators and Reagents; D007328:Insulin / Q000032:analysis; D007515:Islets of Langerhans / Q000737:chemistry; D057230:Limit of Detection; D008297:Male; D051379:Mice
D007202:Indicators and Reagents; D007328:Insulin