1. Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Michael Beltran Robert Lam Bryan Lochman 12/14/07

2. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Lab on a chip Lab on a chip technology will reduce the size of complex experimental setups. Eliminate large, bulky equipment. Move lab experiments to a non-lab environment. Especially useful in biological and medical fields for local use.

3. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Overview Device Overview Theory –Dielectrophoresis (DEP) –DEP cage actuation –Impedance sensing Device Fabrication Previous Devices Results –Parasitic Cages –Particle Concentration Recommendations –Micro-scale device

4. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Device Overview 1cm electrode strips Induced DEP Cages Top conductive sealing layer Integrated actuation and sensing Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab- on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

5. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Theory Dielectrophoresis (DEP) www-dsv.cea.fr/.../Image/Pascal/biopuces_64.jpg

6. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing r – radius E – nonuniform electric field - permittivity of medium Re[K] – Clasius-Mossotti Factor where Theory DEP – Governing Equation

7. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing σ = conductivity of electric field ω = angular frequency of electric field Varying these two variables will alter the permittivity of the particle/medium Theory DEP - Permittivity

8. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing At low frequences: At high frequencies: Polarization Factor (K) can be switched between positive or negative values Theory DEP - Clausius-Mossotti

9. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Buoyancy Force: DEP and Buoyancy: Theory DEP – Vertical Forces Iliescu, C.; Yu, L.; Xu, G.; Tay, F. A Dielectrophoretic Chip With a 3-D Electric Field Gradient, Journal of Microelectromechanical Systems, 2006, 15, 1506-1513

10. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing F DEP = volume (~r 3 ) F viscous = surface (~r 2 ) Smaller particles will move slower Theory DEP – Ratio between DEP, Viscous forces

11. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Progressively alternating electrode signals move particles towards target electrode Provides better sensing of particles Theory DEP – Cage Actuation Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab-on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

12. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing The DEP Cages are able to move toward a target electrode by moving the counter phase signal to the next electrode closer to the target Theory DEP – Cage Actuation Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab-on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

13. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing To measure the concentration of particles impedance sensing is used All electrodes are switched to ground except the sensing electrode The sensing electrode is connected to a transimpedance amplifier Theory Impedance Sensing Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab-on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

14. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Transfer function of the transimpedance amplifier: R M and C M are the resistance and capacitance between the electrode and lid R F and C F are the feedback resistance and capacitance There are two sensing frequency ranges, low and high, if the same signal is used for both DEP cage formation and sensing Theory Impedance Sensing – Transfer Function

15. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Low Frequency –When w<<1/(R M C M ) and w<<1/(R F C F ) the sensing equation is: –The Clausius Mossotti factor at low frequencies, shows that a particle will only be trapped in the DEP cage if its conductivity is lower than the mediums giving rise to : –These two equations show the output voltage will decrease with particles at low frequencies Theory Impedance Sensing at low frequencies

16. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing High Frequency –When w>>1/(R M C M ) and w>>1/(R F C F ) the sensing equation is: –The Clausius Mossotti factor at high frequencies, shows that a particle will only be trapped in the DEP cage if its permittivity is lower than the mediums giving rise to : –These two equations show the output voltage will decrease with particles at high frequencies Theory Impedance Sensing at high frequencies

17. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Original Fabrication No MEMS fabrication methods used Printed Circuit Board (PCB) techniques used to attach electrodes –Silk screened the electrode pattern on to a gold clad board –etched away the uncovered portion –remove the screened resist Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab-on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

18. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Similar DEP Devices CMOS chip for individual cell manipulation 102,400 actuation electrodes (20μm x 20μm) Capability of manipulating 10,000 cells in parallel Lack of integrated sensing technique Manaresi, N.; Romani, A.; Medoro, G.; Altomare, L.; Leonardi, A.; Tartagni, M.; Guerrieri, R. A CMOS Chip for Individual Cell Manipulation and Detection, IEEE Journal of Solid-State Circuits, 2003, 38, 12:2297-2305

19. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Similar DEP Devices Dielectrophoretic Chip With a 3-D Electric Field Gradient Asymmetric 3D electric gradient achieved with specially configured electrodes Thick electrodes integrated into vertical wall structures, thin planar electrodes in bottom substrate Enhanced vertical DEP force (lower voltages and temperatures) Iliescu, C.; Yu, L.; Xu, G.; Tay, F. A Dielectrophoretic Chip With a 3-D Electric Field Gradient, Journal of Microelectromechanical Systems, 2006, 15, 1506-1513

20. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Similar DEP Devices MEMS electrostatic particle transportation system Electrostatic device capable of transporting particles in air Surface modifications performed to reduce adhesive forces Desai, A.; Lee, S-W.; Tai, Y. A MEMS Electrostatic Particle Transportation System. Sensors and Actuators, 1999, 73, 37-44

21. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Parasitic Cages form between the two in-phase electrodes, electrodes 3 and 4 in Figure (a) After actuating the DEP cage, a new parasitic cage will form capturing the slow moving particles (a) (b) Actuate the DEP Cage New parasitic DEP Cage Results Parasitic Cages Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab-on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

22. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Parasitic Cages Results Parasitic Cages Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab-on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

23. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Add intermediate step: Creates a smaller chance slow moving particles will be trapped in the attraction basin of the parasitic cage (a)(b)(c) Results Parasitic Cages – Minimize effects Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab-on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

24. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Reduce space between electrodes: Space between electrodes is nearly too small for particles to fit Only possible using MEMS fabrication techniques due to small spacing Results Parasitic Cages – Minimize effects Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab- on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

25. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Results Modeling Assumptions Cage distribution far too complicated to be modeled at the level of individual particles Assumptions Particle cloud within the DEP cage can be modeled as homogenous Permittivity and Conductivity depend solely on the ratio between the volume of microbeads and suspending medium in the cylinder (distilled water).

26. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Results Signal Processing Fixed pattern noise (FPN) removed by subtracting initial non-cage reading (a) from cage reading, and then addition of average initial reading. Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab-on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

27. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Results Optical observation Polystyrene microbeads, 3.46 µm diameter in H 2 O. 10 Vpp, 100 kHz 4 Concentration cycles Raw data on left Grayscale representation of data on right Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab-on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

28. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Electric field simulation in FEMLAB in a 2-D plane Simulation performed for initial concentration and 4 successive concentration cycles Resistance translated to voltage output with known current Results Polystyrene Microbeads Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab-on-a-Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

29. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Experiment repeated with S. cerevisiae yeast cells in 280-mM mannitol. Mannitol medium used to prevent overheating due to excessive conductivity S. cerevisiae displays pDEP behavior above 200 kHz, electrolysis occurs at less than 30 kHz Experiments performed at 100 kHz Results S. cerevisia Medoro, G.; Manaresi, N.; Leonardi, A.; Altomare, L.; Tartagni, M.; Guerrieri, R. A Lab-on-a- Chip for Cell Detection and Manipulation. IEEE Sensors Journal, 2003, 3, 317-325

30. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Base layer of SiO 2 with photoresist on Silicon Micro-fabrication

31. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Mask pattern, inverted from intended electrode pattern Micro-fabrication

32. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Exposure to light – removal of photoresist. Micro-fabrication

33. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Dry plasma etching – removal of Silicon Oxide Micro-fabrication

34. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Removal of photoresist with acetone Micro-fabrication

35. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Ion implantation of electrode channels Micro-fabrication

36. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Removal of silicon oxide via plasma etching with CF 4 Micro-fabrication

37. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Lap polish of wafer to 50μm thickness Micro-fabrication

38. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Growth of SiO 2 layers, removal from underside. Micro-fabrication

39. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Spin deposition of photoresist and mask placement Micro-fabrication

40. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Exposure to light - removal of photoresist Micro-fabrication

41. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Dry plasma etching – removal of Silicon Oxide Micro-fabrication

42. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Removal of photoresist with acetone Micro-fabrication

43. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations wafer KOH through-etching of silicon wafer Micro-fabrication

44. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Removal of silicon oxide via plasma etching with CF 4 Micro-fabrication

45. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Combination of base electrode layer and reservoir layer Micro-fabrication

46. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Mated with wafer bonding over long electrodes, leaving wire-connection ports exposed Micro-fabrication

47. Michael Beltran Robert Lam Bryan Lochman 12/14/07Three-Dimensional Dielectrophoresis Device with Integrated Actuating and Impedance Sensing Recommendations Mass production on a silicon wafer Glass cover with etched microchannel pattern and common reservoir Micro-fabrication