1. 1 Advisor : Cheng-Hsin Chuang Advisee : Jing-wei Ju Department of Mechanical Engineering & Institute of Nanotechnology, Southern Taiwan University, Tainan, TAIWAN Date ﹕ 2013/05/01 運用介電泳力操控奈米探針 製作免疫抗體陣列晶片

2. Outline Motivation Theory of Dielectrophoresis Simulation of a Non-uniform Electric Field Materials Preparation of nanoprobes Fabrication of the DEP Chip Experiment Setup Procedure Experimental Result Conclusions 2

3. Motivation Many studies utilized micro- and nano-beads for the sensitivity enhancement of biomedical detection due to the large surface- volume ratio. For the diagnosis of cancer, protein analysis is most important. Protein Testing Strips Expensive Time-consuming Only one kind of antibody How to do low-cost and fast detection of multi-antibody on a chip? Multi-antibody array chip

4. Theory of Dielectrophoresis Dielectrophoresis- Motion of a particle produced by the interaction of a Non-uniform electric field with the induced effective dipole moment of the particle. Negative dielectrophoresis Positive dielectrophoresis +V -V Neutral bead Net Force Clausius-Mossotti factor Re(f CM )>0 ; Positive DEP Re(f CM )<0 ; Negative DEP

5. Simulation of a Non-uniform Electric Field Properties/MaterialsMediumSU-8 Density ρ (kg/m 3 )10001194 Conductivity σ (S/m)2E-41E-14 Relative Permittivity ε804 SU-8 Bottom Electrode 50 μm 10 μm 10V PP 10kHz Top Electrode GND 20 μm For Positive DEP Trap NPs

6. Simulation of a Non-uniform Electric Field SU-8 Top Electrode Bottom Electrode SU-8 Top Electrode Bottom Electrode

7. Materials (Preparation of nanoprobes) Nanoprobe: r-Al 2 O 3

8. Materials (Fabrication of the DEP Chip) Double-Sided Tape ITO GlassAntibody-NPs PGS512GlassBottom Electrode MaskSU-8 a. Bottom Electrode Layer Coat MPG512 Layer Deposition Bottom Electrode Layer Etching and Remove PR Expose and Develop Coat SU-8 Layer b. SU-8 Microcavity c. Top Electrode Layer and Chip Bonding Drill on the ITO Glass Bonding Function Generator Ch GND Inlet Outlet

9. Experiment (Setup) Au ITO Inject Outlet

10. Experiment (Procedure)

11. Experimental Result (b) (d) (a) ITO-Electrode 1 2 3 4 5 (c) Au-Electrode 1 2 3 4 5 DEP Without DEP (a) ITO-Electrode(b) (c) Au-Electrode(d) DEP Without DEP 1 2 3 4 5 1 2 3 4 5

12. Experimental Result 2.39 1.88 1.66 2.96 2.55 1.66 1.01 0.53

13. Conclusions 1)We have developed a immunosensor for the immobilization of antibody-NPs in SU-8 microcavities array and the enhancement of fluorescence intensity of immunosensing based on silane-modified scheme and DEP force. 2)According to the experimental results, DEP force not only immobilized the antibody-NPs into the SU-8 microcavities array, but also enhanced the fluorescence intensity of the immunosensing. 3)The transparent electrode not only suit for DEP manipulation of nanoparticles, but also decreases of fluorescent noise for immunosensing. Consequently, it evidenced great potential as a multi- antibody array in a single immune-chip.

14. Future Work Muti-antibody immunoassay chip Sensing the lysate of T24 and R24 cell for special antibodies

15. Thank you for your attention!!