There are highly sensitive analytical techniques for probing cellular and molecular events in very small volumes. The development of microtools for effective sample handling and separation in such volumes remains a challenge. Most devices developed so far use electrophoretic and chromatographic separation methods. We show that forces generated by ac fields under conditions of negative dielectrophoresis (DEP) can also be used. Miniaturized electrode arrays are housed in a microchannel and driven with high-frequency ac. A laminar liquid flow carries particles past the electrodes. Modification of the ac drive changes the particle trajectories. We have handled latex particles of micrometer size and living mammalian cells in a device which consists of the following four elements: a planar funnel which concentrates particles from a 1-mm-wide stream to a beam of about 50-micron width, an aligner which narrows the beam further and acts to break up particle aggregates, a field cage which can be used to trap particles, and a switch which can direct particles into one of two output channels. The electrodes are made from platinum/titanium and indium tin oxide (ITO) on glass substrates. Particle concentration and switching could be achieved for linear flow velocities up to about 10 mm s-1. The combination of this new method with high-performance optical detection offers prospects for miniaturized flow cytometry.

Usually dielectrophoretic and electrorotation measurements are carried out at low ionic strength to reduce electrolysis and heat production. Such problems are minimized in microelectrode chambers. In a planar ultramicroelectrode chamber fabricated by semiconductor technology, we were able to measure the dielectric properties of human red blood cells in the frequency range from 2 kHz to 200 MHz up to physiological ion concentrations. At low ionic strength, red cells exhibit a typical electrorotation spectrum with an antifield rotation peak at low frequencies and a cofield rotation peak at higher ones. With increasing medium conductivity, both electrorotational peaks shift toward higher frequencies. The cofield peak becomes antifield for conductivities higher than 0.5 S/m. Because the polarizability of the external medium at these ionic strengths becomes similar to that of the cytoplasm, properties can be measured more sensitively. The critical dielectrophoretic frequencies were also determined. From our measurements, in the wide conductivity range from 2 mS/m to 1.5 S/m we propose a single-shell erythrocyte model. This pictures the cell as an oblate spheroid with a long semiaxis of 3.3 microns and an axial ratio of 1:2. Its membrane exhibits a capacitance of 0.997 x 10(-2) F/m2 and a specific conductance of 480 S/m2. The cytoplasmic parameters, a conductivity of 0.4 S/m at a dielectric constant of 212, disperse around 15 MHz to become 0.535 S/m and 50, respectively. We attribute this cytoplasmic dispersion to hemoglobin and cytoplasmic ion properties. In electrorotation measurements at about 60 MHz, an unexpectedly low rotation speed was observed. Around 180 MHz, the speed increased dramatically. By analysis of the electric chamber circuit properties, we were able to show that these effects are not due to cell polarization but are instead caused by a dramatic increase in the chamber field strength around 180 MHz. Although the chamber exhibits a resonance around 180 MHz, the harmonic content of the square-topped driving signals generates distortions of electrorotational spectra at far lower frequencies. Possible technological applications of chamber resonances are mentioned.

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We demonstrate that micrometre and sub-micrometre particles can be trapped, aggregated and concentrated in planar quadrupole electrode configurations by positive and negative dielectrophoresis. For particles less than in diameter, concentration is driven by thermal gradients, hydrodynamic effects and sedimentation forces. Liquid streaming is induced by the AC field itself via local heating and results, under special conditions, in vortices which improve the trapping efficiency. Microstructures were fabricated by electron-beam lithography and modified by UV laser ablation. They had typical gap dimensions between 500 nm and several micrometres. The theoretical and experimental results illustrate the basic principles of particle behaviour in ultra-miniaturized field traps filled with aqueous solutions. The smallest single particle that we could stably trap was a Latex bead of 650 nm. The smallest particles which were concentrated in the central part of the field trap were 14 nm in diameter. At high frequencies (in the megahertz range), field strengths up to 56 MV can be applied in the narrow gaps of 500 nm. Further perspectives for microparticle and macromolecular trapping are discussed.

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