Integrated microfluidic device for automated single cell analysis using electrophoretic separation and electrospray ionization mass spectrometry.
A microfabricated fluidic device was developed for the automated real-time analysis of individual cells using capillary electrophoresis (CE) and electrospray ionization-mass spectrometry (ESI-MS). The microfluidic structure incorporates a means for rapid lysis of single cells within a free solution electrophoresis channel, where cellular constituents were separated, and an integrated electrospray emitter for ionization of separated components. The eluent was characterized using mass spectrometry. Human erythrocytes were used as a model system for this study. In this monolithically integrated device, cell lysis occurs at a channel intersection using a combination of rapid buffer exchange and an increase in electric field strength. An electroosmotic pump is incorporated at the end of the electrophoretic separation channel to direct eluent to the integrated electrospray emitter. The dissociated heme group and the alpha and beta subunits of hemoglobin from individual erythrocytes were detected as cells continuously flowed through the device. The average analysis throughput was approximately 12 cells per minute, demonstrating the potential of this method for high-throughput single cell analysis.
Mellors
J Scott
JS
Department of Chemistry, Chapman Hall Room 251, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-3216, USA.
Jorabchi
Kaveh
K
Smith
Lloyd M
LM
Ramsey
J Michael
JM
D002469:Cell Separation; D019075:Electrophoresis, Capillary; D004912:Erythrocytes / Q000166:cytology*; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*