Advisor : Cheng-Hsin Chuang Advisee : Kai-Chieh Chang Department of Mechanical Engineering

1. Biosensors and Bioelectronics 41 (2013) 129–136 On-chip dual detection of cancer biomarkers directly in serum based on self-assembled magnetic bead patterns and quantum dots Xu Yu a,b, He-Shun Xia c, Zuo-Dong Sun a, Yi Lin a,b, Kun Wang c, Jing Yu c, Hao Tang a,c, Dai-Wen Pang a,b, Zhi-Ling Zhang a,b, a Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, and State Key Laboratory of Virology, Wuhan University, Wuhan 430072, PR China bWuhan Institute of Biotechnology, Wuhan 430075, PR China c Hubei Cancer Hospital, Wuhan 430079, PR China Advisor : Cheng-Hsin Chuang Advisee : Kai-Chieh Chang Department of Mechanical Engineering & Institute of Nanotechnology, Southern Taiwan University of Science and Technology, Tainan, TAIWAN Date ﹕2013/11/4

2. Outline • Paper Survey • Experimental • Result and discussion

3. Experimental Fig. 1. Schematic diagrams of an integrated MFCM-Chip and sample loading process. (a) Scheme of Ab-SPMBs loading process. (b) The cross-sectional view of the integrated MFCM-Chip. (c) Scheme of the sample and reagent loading process. (d) Imaging of the nickel pattern arrays in eight parallel channels. (e) Schematic diagrams of the principle of multiplexed detection simultaneously in a MFCM-Chip. The scale bar is 500 mm.

4. Experimental Fig. S5. Optical and fluorescence images of immune complexes. 1.0 μg/mL CEA was added into two 1.5 mL eppendorf tube with the anti-CEA-SPMBs and incubated for 30 min. (A) (B) Without biotin-anti-CEA adding into the immunoreaction. (C) (D) With biotin-anti-CEA (10.0 μg/mL) adding into the immunoreaction. SA-QDs (1 μL, 1 × 10-6 mol/L) were added into two tubes and incubated for 15 min. The scale bars are 20 μm. Fig. S4. (A) (B) Optical and fluorescence images of SPMBs. (C) (D) Optical and fluorescence images of the complexes which were the products of the immunoreaction of the anti-AFP-SPMBs and anti-mouse IgG-FITC. The scale bars are 20 μm.

5. Experimental

6. Fig. S9. Simulation of the absolute value of the multiplication of magnetic flux density and the magnetic field gradient ((B⋅∇) B). The scale bar is 50 μm. Fig. S8. A MFCM-Chip was used to detection of 200 ng/mL CEA for 16 times.

7. Results and discussion Fig. 3. Multiplexed detection in a MFCM-Chip. (A) Nickel pattern arrays under eight parallel branch fluid channels. (B) Different captured Ab-SPMBs in the branch channels. (C) Fluorescence image of detection of multiple target Ags. From channel 1 to channel 8 to detect AFP, none, rabbit IgG, CEA, rabbit IgG, AFP, none and CEA, respectively. (D) Fluorescence image after several minutes’ excitement by blue light. The scale bars are 200 mm.

9. Fig. 5. (a) Fluorescence images of detection of different concentrations of CEA. (b) Calibration curves of simultaneous detection of CEA and AFP from 10.0 ng/mL to 400.0 ng/mL at the exposure time of 400 ms. (c) Calibration curves of detection of CEA and AFP from 200.0 ng/mL to 2000.0 ng/mL with the exposure time of 200 ms. (d) Normalized calibration curves of detection of CEA and AFP from 10.0 ng/mL to 2000.0 ng/mL. (e) The linear relationship between fluorescence intensity and the concentration of AFP and CEA in the range from 10.0 ng/mL to 800.0 ng/mL (R2 ¼0.982 and 0.998, respectively). The scale bar is 50 mm.

10. Results and discussion Fig. 6. Simultaneous detection of AFP and CEA in clinical serum samples. The values of background fluorescence intensity were not deducted.

11. Thank you for your attention