Frequency discretization in dielectrophoretic assisted cell sorting arrays to isolate neural cells.

We present an automated dielectrophoretic assisted cell sorting (DACS) device for dielectric characterization and isolation of neural cells. Dielectrophoretic (DEP) principles are often used to develop cell sorting techniques. Here we report the first statistically significant neuronal sorting using DACS to enrich neurons from a heterogeneous population of mouse derived neural stem/progenitor cells (NSPCs) and neurons. We also study the dielectric dispersions within a heterogeneous cell population using a Monte-Carlo (MC) simulation. This simulation model explains the trapping behavior of populations as a function of frequency and predicts sorting efficiencies. The platform consists of a DEP electrode array with three multiplexed trapping regions that can be independently activated at different frequencies. A novel microfluidic manifold enables cell sorting by trapping and collecting cells at discrete frequency bands rather than single frequencies. The device is used to first determine the percentage of cells trapped at these frequency bands. With this characterization and the MC simulation we choose the optimal parameters for neuronal sorting. Cell sorting experiments presented achieve a 1.4-fold neuronal enrichment as predicted by our model.

[1]  Michael P Hughes,et al.  Dielectrophoresis-activated multiwell plate for label-free high-throughput drug assessment. , 2008, Analytical chemistry.

[2]  Li-Ling Chak,et al.  Human Wharton’s Jelly Stem Cells Have Unique Transcriptome Profiles Compared to Human Embryonic Stem Cells and Other Mesenchymal Stem Cells , 2011, Stem Cell Reviews and Reports.

[3]  S. Quake,et al.  Monolithic microfabricated valves and pumps by multilayer soft lithography. , 2000, Science.

[4]  R. Pethig,et al.  Electrorotation and dielectrophoresis , 1999, Parasitology.

[5]  Abraham P Lee,et al.  Dual frequency dielectrophoresis with interdigitated sidewall electrodes for microfluidic flow‐through separation of beads and cells , 2009, Electrophoresis.

[6]  Hywel Morgan,et al.  Negative DEP traps for single cell immobilisation. , 2009, Lab on a chip.

[7]  Byungkyu Kim,et al.  Separation of malignant human breast cancer epithelial cells from healthy epithelial cells using an advanced dielectrophoresis-activated cell sorter (DACS) , 2009, Analytical and bioanalytical chemistry.

[8]  J. Voldman Electrical forces for microscale cell manipulation. , 2006, Annual review of biomedical engineering.

[9]  Thomas Braschler,et al.  A unified approach to dielectric single cell analysis: impedance and dielectrophoretic force spectroscopy. , 2010, Lab on a chip.

[10]  Satya Prakash,et al.  Oral Microencapsulated Live Saccharomyces cerevisiae Cells for Use in Renal Failure Uremia: Preparation and In Vivo Analysis , 2010, Journal of biomedicine & biotechnology.

[11]  Saeid Nahavandi,et al.  Dielectrophoretic-activated cell sorter based on curved microelectrodes , 2010 .

[12]  R. Pethig,et al.  Dielectrophoresis: A Review of Applications for Stem Cell Research , 2010, Journal of biomedicine & biotechnology.

[13]  F. Gage,et al.  Mammalian neural stem cells. , 2000, Science.

[14]  Noo Li Jeon,et al.  Unique Dielectric Properties Distinguish Stem Cells and Their Differentiated Progeny , 2008, Stem cells.

[15]  H. Morgan,et al.  The dielectrophoretic and travelling wave forces generated by interdigitated electrode arrays: analytical solution using Fourier series , 2001 .

[16]  A. Irimajiri,et al.  A dielectric theory of "multi-stratified shell" model with its application to a lymphoma cell. , 1979, Journal of theoretical biology.

[17]  Y. Huang,et al.  Differences in the AC electrodynamics of viable and non-viable yeast cells determined through combined dielectrophoresis and electrorotation studies. , 1992, Physics in medicine and biology.

[18]  K R Foster,et al.  Electrorotation and levitation of cells and colloidal particles. , 1992, Biophysical journal.

[19]  Peter R C Gascoyne,et al.  Enrichment of putative stem cells from adipose tissue using dielectrophoretic field-flow fractionation. , 2008, Lab on a chip.

[20]  Ronald Pethig,et al.  Dielectrophoretic studies of the activation of human T lymphocytes using a newly developed cell profiling system , 2002, Electrophoresis.

[21]  Paul H. Bessette,et al.  Marker-specific sorting of rare cells using dielectrophoresis. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[22]  T. Tsong,et al.  Dielectrophoresis and electrorotation of neurospora slime and murine myeloma cells. , 1991, Biophysical journal.

[23]  J. Voldman,et al.  An equilibrium method for continuous-flow cell sorting using dielectrophoresis. , 2008, Analytical chemistry.

[24]  R. Pethig,et al.  Dielectrophoretic separation and enrichment of CD34+cell subpopulation from bone marrow and peripheral blood stem cells , 1995, Medical and Biological Engineering and Computing.

[25]  R. Pethig,et al.  Dielectrophoretic separation of cells: Continuous separation , 1995, Biotechnology and bioengineering.

[26]  Cengiz S. Ozkan,et al.  Separation of individual neurons using dielectrophoretic alternative current fields , 2004, Journal of Neuroscience Methods.

[27]  Michael P Hughes,et al.  Extraction of dielectric properties of multiple populations from dielectrophoretic collection spectrum data , 2005, Physics in medicine and biology.

[28]  Gideon Rechavi,et al.  Donor-Derived Brain Tumor Following Neural Stem Cell Transplantation in an Ataxia Telangiectasia Patient , 2009, PLoS medicine.

[29]  Patrick S Daugherty,et al.  Multitarget dielectrophoresis activated cell sorter. , 2008, Analytical chemistry.

[30]  Thomas B. Jones,et al.  Multipolar dielectrophoretic and electrorotation theory , 1996 .

[31]  Thomas Braschler,et al.  Continuous separation of cells by balanced dielectrophoretic forces at multiple frequencies. , 2008, Lab on a chip.

[32]  W. Stahel,et al.  Log-normal Distributions across the Sciences: Keys and Clues , 2001 .

[33]  P. Smith,et al.  Electrokinetic measurements of membrane capacitance and conductance for pancreatic beta-cells. , 2005, IEE proceedings. Nanobiotechnology.