Fast-lysis cell traps for chemical cytometry.
Electrically addressable cell traps were integrated with capillary electrophoresis for the analysis of the contents of single adherent cells. Electrodes composed of indium tin oxide were patterned on a glass surface followed by formation of topographical cell traps using 1002F photoresist. Single cells trapped in the holes could be lysed in less than 66 ms by applying a brief electric field (10 ms) across the electrode beneath the cell and the ground electrode placed in the aqueous media above the cell traps. The gas formed during cell lysis remained localized within the cavity formed by the 1002F photoresist. The retention of the gas in the cell trap enabled the cell traps to be coupled to an overlying capillary without blockage of the capillary. Single cells cultured in the traps were loaded with fluorescein and Oregon Green and then electrically lysed. By simultaneous application of an electric field to the capillary, the cell's contents were loaded into the capillary and electrophoretically separated. Orgeon Green and fluorescein from a single cell were fully resolved in less than two minutes. The use of a single patterned electrode beneath the 1002F cell trap yielded a simple easily fabricated design that was robust when immersed in aqueous solutions. Moreover, the design can easily be scaled up to create arrays of adherent cells for serial analyses using a single capillary or for parallel analysis by mating to an array of capillaries. Enhancing the rate of analysis of single adherent cells would enable a greater understanding of cellular physiology.
Marc
Paul J
PJ
Department of Biomedical Engineering, University of California, Irvine, California 92697, USA.
Sims
Christopher E
CE
Bachman
Mark
M
Li
G P
GP
Allbritton
Nancy L
NL
D000818:Animals; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D004566:Electrodes; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods*; D019793:Fluorescein / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D051381:Rats; D012680:Sensitivity and Specificity; D013499:Surface Properties
D005456:Fluorescent Dyes; D019793:Fluorescein