Interface of an array of five capillaries with an array of one-nanoliter wells for high-resolution electrophoretic analysis as an approach to high-throughput chemical cytometry.

Analytical Chemistry
Anna BoardmanNorman J Dovichi

Abstract

We report a system that allows the simultaneous aspiration of one or more cells into each of five capillaries for electrophoresis analysis. A glass wafer was etched to create an array of 1-nL wells. The glass was treated with poly(2-hydroxyethyl methacrylate) to control cell adherence. A suspension of formalin-fixed cells was placed on the surface, and cells were allowed to settle. The concentration of cells and the settling time were chosen so that there was, on average, one cell per well. Next, an array of five capillaries was placed so that the tip of each capillary was in contact with a single well. A pulse of vacuum was applied to the distal end of the capillaries to aspirate the content of each well into a capillary. Next, the tips of the capillaries were placed in running buffer and potential was applied. The cells lysed upon contact with the running buffer, and fluorescent components were detected at the distal end of the capillaries by laser-induced fluorescence. The electrophoretic separation efficiency was outstanding, generating over 750,000 theoretical plates (1,800,000 plates/m). In this example, AtT-20 cells were used that had been treated with TMR-G(M1). The cells were allowed to metabolize this substrate into a...Continue Reading

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Citations

Apr 20, 2011·Analytical Chemistry·Yuqing LinAndrew G Ewing
Jan 31, 2012·The Analyst·Christine Cecala, Jonathan V Sweedler
Jan 9, 2013·Luminescence : the Journal of Biological and Chemical Luminescence·Chunxiu Xu, Longfei Cai
Mar 27, 2013·Analytical Chemistry·Alexandra J DickinsonNancy L Allbritton

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