Measurement of the real part of the Clausius-Mossotti factor of dielectrophoresis for Brownian particles.

A method is proposed for measuring the real part of the Clausius-Mossotti factor ( Kr ) of dielectrophoresis for Brownian particles based on a solution of the Smoluchowski equation using a designed polydimethysilloxane microchannel with planar hyperbolic electrodes on its glass substrate. An approximate two-dimensional spring-like dielectrophoretic force is generated in the device, and the data necessarily measured is the time evolution of the in-plane particle displacement undergoing confined Brownian motion. Validity of the measurement was checked against the zeta potentials in the literature based on the classical theory of surface conductance using polystyrene particles of size of one micron. As the dielectrophoretic force depends on Kr , which is usually unknown for bio-particles and some engineered particles, and is seldom measured; this study is important from the academic point of view and could be helpful for the manipulation and characterization of sub-micron particles using dielectrophoresis. Extension of the method to the measurement of permanent dipole moment and total polarizability of particle was developed theoretically and discussed by incorporating an optical tweezer into the present device.

Measurement of the Imaginary Part of the Clausius-Mossotti Factor of Particle/Cell via Dual Frequency Electrorotation.

A simple and inexpensive method using planar electrodes was proposed for the measurement of the imaginary part of the Clausius-Mossotti factor, K i , of particle/cell for electrorotation (ER) and travelling wave dielectrophoresis (twDEP). It is based on the balance between the dielectrophoretic and viscous torques on a particle undergoing ER subject to dual frequency operation in an ER chamber. A four-phase ac voltage signal with a given frequency is applied for generating ER for measurement, and another two-phase signal is applied at a selected frequency for generating a negative dielectrophoretic force for confining the particle motion, instead of using laser tweezer or three-dimensional electrodes in the literature. Both frequencies can be applied to the same electrodes in a four-electrode ER system and to alternative different electrodes in an eight-electrode ER system, and both systems are capable for providing accurate measurement. The measurements were validated by comparing with the theoretical result using sephadex particles in KCl solution, and with the existing experimental results for various human cancer cells in medium with conductivity from 0.01-1.2 S/m, using ER with optical tweezer and dual frequency twDEP. Contrast between the ER and the twDEP methods (the current two available methods) was discussed and commented. The present method could provide measurement for wider frequency range and more accurate result near K i = 0, in comparison with the results using the twDEP method. However, the twDEP method could perform much more rapid measurement. Detailed forces and torque were calculated inside the ER chamber for understanding the physics and assessing the characteristics of the dual frequency ER method. This study is of academic interest as the torque in ER and the force in twDEP can be calculated only when K i is known. It also finds biomedical applications as the K i -spectra can be served as physical phenotypes for different cells, and can be applied for deriving dielectric properties of cells.

A Continuous Flow-through Microfluidic Device for Electrical Lysis of Cells.

In contrast to the delicate 3D electrodes in the literature, a simple flow-through device is proposed here for continuous and massive lysis of cells using electricity. The device is essentially a rectangular microchannel with a planar electrode array built on its bottom wall, actuated by alternating current (AC) voltages between neighboring electrodes, and can be incorporated easily into other biomedical systems. Human whole blood diluted 10 times with phosphate-buffered saline (about 6 108 cells per mL) was pumped through the device, and the cells were completely lysed within 7 s after the application of a 20 V peak-to-peak voltage at 1 MHz, up to 400 µL/hr. Electric field and Maxwell stress were calculated for assessing electrical lysis. Only the lower half-channel was exposed to an electric field exceeding the irreversible threshold value of cell electroporation (Eth2), suggesting that a cross flow, proposed here primarily as the electro-thermally induced flow, was responsible for bringing the cells in the upper half-channel downward to the lower half-channel. The Maxwell shear stress associated with Eth2 was one order of magnitude less than the threshold mechanical stresses for lysis, implying that an applied moderate mechanical stress could aid electrical lysis.

Three dimensional electrode array for cell lysis via electroporation.

Microfabricated devices for cell lysis have demonstrated many advantages over conventional approaches. Among various design of microdevices that employ electroporation for cytolysis, most utilize Ag/AgCl wires or 2D planar electrodes. Although, simple in fabrication the electric field generated by 2D electrodes decays exponentially, resulting in rather non-uniform forcing on the cell membrane. This paper investigates the effect of electric field generated by 3D cylindrical electrodes to perform cell lysis via electroporation in a microfluidic platform, and compared with that by 2D design. Computational results of the electric field for both 2D and 3D electrode geometries showed that the 3D configuration demonstrated a significantly higher effective volume ratio-volume which electric field is sufficient for cell lysis to that of net throughflow volume. Hence, the efficacy of performing cell lysis is substantially greater for cells passing through 3D than 2D electrodes. Experimentally, simultaneous multi-pores were observed on leukocytes lysed with 3D electrodes, which is indicative of enhanced uniformity of the electric field generated by 3D design. Additionally, a single row of 3D electrode demonstrated a substantially higher lysing percentage (30%) than that of 2D (8%) under that same flow condition. This work should aid in the design of electrodes in performing cell lysis via electroporation.