A real-time multiple-cell tracking platform for dielectrophoresis (DEP)-based cellular analysis

There is an increasing demand from biosciences to develop new and efficient techniques to assist in the preparation and analysis of biological samples such as cells in suspension. A dielectrophoresis (DEP)-based characterization and measurement technique on biological cells opens up a broader perspective for early diagnosis of diseases. An efficient real-time multiple-cell tracking platform coupled with DEP to capture and quantify the dynamics of cell motion and obtain cell viability information is presented. The procedure for tracking a single DEP-levitated Canola plant protoplast, using the motion-based segmentation algorithm hierarchical adaptive merge split mesh-based technique (HAMSM) for cell identification, has been enhanced for identifying and tracking multiple cells. The tracking technique relies on the deformation of mesh topology that is generated according to the movement of biological cells in a sequence of images that allows the simultaneous extraction of the biological cell from the image and the associated motion characteristics. Preliminary tests were conducted with yeast cells and then applied to a cancerous cell line subjected to DEP fields. Characteristics, such as cell count, velocity and size, were individually extracted from the tracked results of the cell sample. Tests were limited to eight yeast cells and two cancer cells. A performance analysis to assess tracking accuracy, computational effort and processing time was also conducted. The tracking technique employed on model intact cells in DEP fields proved to be accurate, reliable and robust.

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

[2]  Ronald Pethig,et al.  Computer-aided analyses of electric fields used in electrorotation studies , 1994 .

[3]  Karan V. I. S. Kaler,et al.  Integrated planar concentric ring dielectrophoretic (DEP) levitator , 2002 .

[4]  Bilge Gunsel,et al.  Two-dimensional mesh-based visual-object representation for interactive synthetic/natural digital video , 1998, Proc. IEEE.

[5]  H. P. Schwan,et al.  Surface conductance and other properties of latex particles measured by electrorotation , 1987 .

[6]  Thomas B. Jones,et al.  Active feedback‐controlled dielectrophoretic levitation , 1986 .

[7]  Biological cell motion tracking in dielectrophoresis (DEP) levitation feedback control system , 2002, IEEE CCECE2002. Canadian Conference on Electrical and Computer Engineering. Conference Proceedings (Cat. No.02CH37373).

[8]  Masao Washizu,et al.  Applications of electrostatic stretch-and-positioning of DNA , 1993, Conference Record of the 1993 IEEE Industry Applications Conference Twenty-Eighth IAS Annual Meeting.

[9]  Berthold K. P. Horn,et al.  Determining Optical Flow , 1981, Other Conferences.

[10]  Sophia Adamia,et al.  A combined dielectrophoresis, traveling wave dielectrophoresis and electrorotation microchip for the manipulation and characterization of human malignant cells. , 2004, Journal of microbiological methods.

[11]  Aria Nosratinia,et al.  New kernels for fast mesh-based motion estimation , 2001, IEEE Trans. Circuits Syst. Video Technol..

[12]  Esther Guohua Cen.  An integrated microchip for cellular analysis and manipulation , 2003 .

[13]  G. Fuhr,et al.  A new microsystem for automated electrorotation measurements using laser tweezers. , 2000, Biochimica et biophysica acta.

[14]  Peter R. C. Gascoyne,et al.  Dielectrophoresis-based sample handling in general-purpose programmable diagnostic instruments , 2004, Proceedings of the IEEE.

[15]  Hywel Morgan,et al.  Dielectrophoretic separation of nano-particles , 1997 .

[16]  Ronald Pethig,et al.  The dielectrophoretic levitation of latex beads, with reference to field-flow fractionation , 1997 .

[17]  Steven R. Visuri,et al.  Microfluidic sample preparation for immunoassays , 2001, MOEMS-MEMS.

[18]  H. A. Pohl,et al.  Dielectrophoresis: The Behavior of Neutral Matter in Nonuniform Electric Fields , 1978 .

[19]  U. Zimmermann,et al.  Dielectric spectroscopy of Schizosaccharomyces pombe using electrorotation and electroorientation. , 2001, Biochimica et biophysica acta.

[20]  R. Pethig,et al.  Use of dielectrophoretic collection spectra for characterizing differences between normal and cancerous cells , 1992, Conference Record of the 1992 IEEE Industry Applications Society Annual Meeting.

[21]  Daniel Mietchen,et al.  Automated dielectric single cell spectroscopy - temperature dependence of electrorotation , 2002 .

[22]  H Morgan,et al.  Measurement of Bacterial Flagellar Thrust by Negative Dielectrophoresis , 1999, Biotechnology progress.

[23]  H Zola.  Immunological applications of flow cytometry. , 2000, Journal of immunological methods.

[24]  G. Harpavat.  Magnetostatic forces on a chain of spherical beads in a nonuniform magnetic field , 1974 .

[25]  Jerry L. Potter,et al.  Velocity as a Cue to Segmentation , 1975, IEEE Transactions on Systems, Man, and Cybernetics.

[26]  Giovanni De Gasperis,et al.  Automated electrorotation: dielectric characterization of living cells by real-time motion estimation , 1998 .

[27]  A. Murat Tekalp,et al.  Bayesian segmentation of MR images using 3D Gibbsian priors , 1993, Electronic Imaging.

[28]  Anil K. Jain,et al.  Displacement Measurement and Its Application in Interframe Image Coding , 1981, IEEE Trans. Commun..

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

[30]  Magdy A. Bayoumi,et al.  A Low Power Architecture for HASM Motion Tracking , 2004, J. VLSI Signal Process..

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

[32]  P. Anandan,et al.  A computational framework and an algorithm for the measurement of visual motion , 1987, International Journal of Computer Vision.

[33]  Ching Y. Suen,et al.  A recursive thresholding technique for image segmentation , 1998, IEEE Trans. Image Process..

[34]  F F Becker,et al.  Cell separation on microfabricated electrodes using dielectrophoretic/gravitational field-flow fractionation. , 1999, Analytical chemistry.

[35]  Nicolas Kalogerakis,et al.  A micromachined DEP cell filtration device , 1996, IAS '96. Conference Record of the 1996 IEEE Industry Applications Conference Thirty-First IAS Annual Meeting.

[36]  Sankar K. Pal,et al.  A review on image segmentation techniques , 1993, Pattern Recognit..

[37]  R Pethig,et al.  Automatic cell electrorotation measurements: studies of the biological effects of low-frequency magnetic fields and of heat shock. , 1998, Physics in medicine and biology.

[38]  Ronald Pethig,et al.  Theoretical and experimental investigations of the interdependence of the dielectric, dielectrophoretic and electrorotational behaviour of colloidal particles , 1993 .