Dielectrophoretic separation of mouse melanoma clones

Biomicrofluidics (Impact Factor: 3.36). 06/2010; 4(2). DOI: 10.1063/1.3447702
Source: PubMed


Dielectrophoresis (DEP) is employed to differentiate clones of mouse melanoma B16F10 cells. Five clones were tested on microelectrodes. At a specific excitation frequency, clone 1 showed a different DEP response than the other four. Growth rate, melanin content, recovery from cryopreservation, and in vitro invasive studies were performed. Clone 1 is shown to have significantly different melanin content and recovery rate from cryopreservation. This paper reports the ability of DEP to differentiate between two malignant cells of the same origin. Different DEP responses of the two clones could be linked to their melanin content.

Download full-text


Available from: Stephen Beebe, Feb 10, 2014
  • Source
    • "In most cases, several different experimental conditions are tested and the purity and recovery of the results are analyzed to determine the optimum conditions. Two notable exceptions are the work by Gascoyne et al., [28] and Sabuncu and Beskok [29]. They have developed a separability parameter to measure the expected difference in the pDEP and nDEP responses of the target cells. "
    [Show abstract] [Hide abstract]
    ABSTRACT: We present a method to quantify and enhance separation of binary cells mixture in the microfluidic device using high frequency dielectrophoresis (>20 MHz). At these frequencies, the DEP response depends primarily on the dielectric properties of the cytoplasm. In order to achieve efficient separation, there must be a difference in the intrinsic dielectric properties of the populations to be sorted. For algae cells, the shift in high frequency dielectrophoresis response during lipid accumulation can be used as a basis of separation. We defined a separability parameter based on the expected difference in the dielectrophoresis responses of the algae cells. Chlamydomonas reinhardtii cells were cultured in regular media and then the same cells were cultured under nitrogen-free conditions to accumulate neutral (non-polar) lipids. Separability of microalgae cells with different lipid content via high frequency dielectrophoresis were investigated by a thin needle shaped electrodes patterned by standard photolithographic and wet etching procedures. Experimental separability factors were measured by estimation of relative lipid content with BODIPY 505/515 fluorescence dye and calculating the area-weighted intensity average of fluorescent images. Theoretical separability parameter was calculated using analytical analysis of single shell model by MATLAB. Theoretical and experimental separability parameters, as tools to determine the optimal separation method, were calculated for microalgae cells with different lipid content. This objective function was maximized in the range of 35-45 MHz for C. reinhardtii cells after 21 days of lipid accumulation in a static separation. In order to design a continuous cell sorter device, the theoretical separation factor was maximized based on differences in the magnitude or the direction of the DEP force.
    ASME 2015 Technical Conference and Exhibition on Packaging and Integration of Electronic and Photonic Microsystems and ASME 2015 12th International Conference on Nanochannels, Microchannels, and Minichannels InterPACKICNMM2015, San Francisco, California, USA; 07/2015
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Selection of particles or cells of specific shapes from a complex mixture is an essential procedure for various biological and industrial applications, including synchronization of the cell cycle, classification of environmental bacteria, and elimination of aggregates from synthesized particles. Here, we investigate the separation behaviors of nonspherical and spherical particles∕cells in the hydrodynamic filtration (HDF) scheme, which was previously developed for continuous size-dependent particle∕cell separation. Nonspherical particle models were prepared by coating the hemisphere of spherical polymer particles with a thin Au layer and by bonding the Janus particles to form twins and triplets resembling dividing and aggregating cells, respectively. High-speed imaging revealed a difference in the separation behaviors of spherical and nonspherical particles at a branch point; nonspherical particles showed rotation behavior and did not enter the branch channel even when their minor axis was smaller than the virtual width of the flow region entering the branch channel, w(1). The confocal-laser high-speed particle intensity velocimetry system visualized the flow profile inside the HDF microchannel, demonstrating that the steep flow-velocity distribution at the branch point is the main factor causing the rotation behavior of nonspherical particles. As applications, we successfully separated spherical and nonspherical particles with various major∕minor lengths and also demonstrated the selection of budding∕single cells from a yeast cell mixture. We therefore conclude that the HDF scheme can be used for continuous shape-based particle∕cell separation.
    Biomicrofluidics 04/2011; 5(2):24103. DOI:10.1063/1.3580757 · 3.36 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Over the past decade, dielectrophoresis (DEP) has evolved into a powerful, robust and flexible method for cellular characterization, manipulation, separation and cell patterning. It is a field with widely varying disciplines, as it is quite common to see DEP integrated with a host of applications including microfluidics, impedance spectroscopy, tissue engineering, real-time PCR, immunoassays, stem-cell characterization, gene transfection and electroporation, just to name a few. The field is finally at the point where analytical and numerical polarization models can be used to adequately describe and characterize the dielectrophoretic behavior of cells, and there is ever increasing evidence demonstrating that electric fields can safely be used to manipulate cells without harm. As such, DEP is slowly making its way into the biological sciences. Today, DEP is being used to manipulate individual cells to specific regions of space for single-cell assays. DEP is able to separate rare cells from a heterogeneous cell suspension, where isolated cells can then be characterized and dynamically studied using nothing more than electric fields. However, there is need for a critical report to integrate the many new features of DEP for cellular applications. Here, a review of the basic theory and current applications of DEP, specifically for cells, is presented.
    Electrophoresis 09/2011; 32(18):2466-87. DOI:10.1002/elps.201100060 · 3.03 Impact Factor
Show more