Coaxial flow system for chemical cytometry.
Over the past decade, chemical cytometry performed by capillary electrophoresis (CE) has become increasingly valuable as a bioanalytical tool to quantify analytes from single cells. However, extensive use of CE-based chemical cytometry has been hindered by the relatively low throughput for the analysis of single adherent cells. In order to overcome the low throughput of CE-based analysis of adherent cells and increase its utility in evaluating cellular attributes, new higher throughput methods are needed. Integration of a coaxial buffer exchange system with CE-based chemical cytometry increased the rate of serial analyses of cells. In the designed system, fluid flow through a tube coaxial to the separation capillary was used to supply electrophoretic buffer to the capillary. This sheath or coaxial fluid was turned off between analysis of cells and on during cell sampling and electrophoresis. Thus, living cells were not exposed to the nonphysiologic electrophoretic buffer prior to lysis. Key parameters of the system such as the relative capillary-sheath positions, buffer flow velocities, and the cell chamber design were optimized. To demonstrate the utility of the system, rat basophilic leukemic cells loaded with Oregon green and fluorescein were serially lysed and loaded into a capillary. Separation of the contents of 20 cells at a rate of 0.5 cells/min was demonstrated.
Marc
Paul J
PJ
Department of Biomedical Engineering, University of California, Irvine, California 92697, USA.
Sims
Christopher E
CE
Allbritton
Nancy L
NL
D002021:Buffers; D019075:Electrophoresis, Capillary / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet
D002021:Buffers