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title page
Contents
1. Introduction 11
1.1. Conducting polymer 12
1.2. PEDOT and PEDOT/PSS dispersion 14
1.3. Conducting polymers for enzyme immobilization 17
1.4. Glucose sensor using glucose oxidase 20
1.5. Michaelis-Menten theory 23
1.6. Non-conducting polymers as a permselective membrane 25
1.7. Aim of the study 26
2. Experimental 28
2.1. Materials & equipments 28
2.2. Preparation of capped microstructure 29
2.3. Enzyme loading 32
2.4. Scanning electron microscope imaging 33
2.5. Gold treatment 33
2.6. Electrochemical measurements 33
2.6.1. Preparation of the capped structure 33
2.6.2. Determination of glucose concentration 34
2.7. Interference and stability tests 35
3. Results and discussions 36
3.1. Formation of micro-tubule arrays 36
3.2. Capping of the opening 46
3.3. Inclusion and immobilization of electroactive materials 54
3.4. Response of microtubule array electrode to H₂O₂ 55
3.5. Effect of capping on response 66
3.6. Effect of polymerization cycles for capping on response 68
3.7. Effect of monitoring potential on response 71
3.8. Effect of enzyme concentration on response 72
3.9. Interference tests 73
3.10. Analytical performances 78
4. Conclusions 84
5. REFERENCES 86
ABSTRACT 93
국문초록 95
Table 1.1. Well-known conducting polymers and their conductivities 13
Table 3.1. Summary of the analytical performances 80
Table 3.2. Examples of glucose sensor based on GOx immobilization 80
Figure 1.1. Structure of PEDOT(poly(3,4-ethylenedioxythiophene)). 15
Figure 1.2. Structure of PEDOT/PSS. 16
Figure 1.3. Structure of glucose oxidase. 22
Figure 1.4. Molecular structure and electrochemical reaction of FAD. 22
Figure 1.5. Schematic diagram of glucose sensing in the capped PEDOT tubule. 27
Figure 2.1. A cross-sectional view of the measuring cell. WE, CE, RE for working, counter, reference electrode, respectively. 35
Figure 3.1. SEM images of polycarbonate membrane with 1.2 ㎛ and 0.05 μm pores. 36
Figure 3.2. Cyclic voltammograms for making tubules in PC membrane (pore size=1.2 μm). Scan rate = 50 mV/s. Monomer solutions are 0.1 M EDOT/0.1 M LiClO₄/acetonitrile with different water content. 15 cycles.... 39
Figure 3.3. The first 6 segments of each cyclic voltammogram in Figure 3.2. Arrow : increasing water content. 40
Figure 3.4. Cyclic voltammograms of tubule-formed membrane (a) in 0.1 M LiClO₄/acetonirtile and (b) in 0.1 M LiClO₄/water, respectively. Scan rate = 50 mV/s. 40
Figure 3.5. SEM images of tubules prepared with different number of potential cycling. 42
Figure 3.6. SEM images of tubules prepared from monomer solutions with different water content 43
Figure 3.7. SEM images of tubules before and after immersing in water for 15 minutes. The tubules were made by cycling the potential 15 times from 0.3 to 1.2 V in 0.1 M EDOT/0.1 M LiClO₄/acetonitrile. 45
Figure 3.8. CV of EDOT polymerization in 0.1 M EDOT/0.1 M LiClO₄/acetonitrile/water(water content=50%) after covering the capping conducting composite film with a membrane of 0.05 μm pores.... 47
Figure 3.9. Surface morphology of the composite film on the PEDOT tubule after elecropolymerization in the 0.1 M EDOT/0.1 M LiClO₄/acetonitrile solution(water content=50%).... 49
Figure 3.10. Changes of surface morphologies of capping film without electropolymerization of EDOT onto it. It was treated with water for 5 minutes. The left is before water treatment and the right is after water treatment. 50
Figure 3.11. SEM image of the tubule bottom after detaching it from ITO glass. 51
Figure 3.12. Schematic diagram showing the slicing the micro structure with a surgical blade after the capping process. 52
Figure 3.13. SEM images of the capped microstructure before (a) and after (b) removing the template and supporting PC membrane. The left part of the whole structure was previously cut at an angle of 45 degree to the... 53
Figure 3.14. CVs of the micro structured electrode in 0.1 M KCI aqueous solution after capping with the composite and PEDOT. 0, 1, 5, 20 mM K₃Fe(CN)6 was allowed to enter the tubule before capping with the...(이미지참조) 55
Figure 3.15. Comparison of responses to H₂O₂ between gold-treated (b,c) and naive tubule electrode (a). (b) is the electrode from gold-sputtered membrane and (c) is the electrochemically gold-deposited electrode.... 56
Figure 3.16. SEM images(a,b) and EDAX data(c,d) of PC membrane(1.2 ㎛ pore size) sputtered with gold. (a), (c) from gold-sputtered surface and (b), (d) from the opposite side. 57
Figure 3.17. CVs of microtubule array electrode in 0.5 M H₂SO₄ with(thick line) and without (thin line) 1 mM HAuC1₄. Scan rate = 50 mV/s. 59
Figure 3.18. SEM image(a) and EDAX data(b) of the microtubule electrode array after electrochemical deposition of HAuCl₄ solution by CV (scan rate = 50 mV/s). 61
Figure 3.19. CVs of microtubule array electrode in 1 mM HAuCl₄/0.5 M H₂SO₄ solution (scan rate = 10 mV/s). 62
Figure 3.20. SEM image of the microtubule electrode array after electrochemical deposition of HAuCl₄ solution by CV (scan rate = 10 mV/s). 62
Figure 3.21. Chronoamperograms of microtubule array electrode for gold deposition. Inset is the zoom-in of 0-10 s area. Duration time = 10s, 30s, 100s in 1 mM HAuCl₄/0.5 M H₂SO₄ and 100s in 0.5 M H₂SO₄... 63
Figure 3.22. SEM images of the microtubule electrode array after electrochemical deposition of HAuCl₄ solution by CA (duration time=10 sec (a), 100 sec(b)). 64
Figure 3.23. Amperometric responses of microtubules to 1 mM glucose (in gold sputtered membrane vs. naive one) (E=0.7 V vs. Ag/AgCl, counter=Pt plate) 66
Figure 3.24. Responses of the enzyme electrodes to glucose with(circle) and without(rectangle) capping procedure. (E=0.7 V vs. Ag/AgCl, counter=Pt plate). GOx solution of 50 mg/ml was treated. 67
Figure 3.25. Responses of the enzyme electrodes capped with different number of potential cycling. n is the number of potential cycling from 0.3V to 1.2 V for electrochemical polymerization of EDOT.... 69
Figure 3.26. SEM images of the surface of microstructure after removing 0.05 ㎛ supporting membrane. Number of polymerization cycle for capping = 1(a), 3(b), 5(c), 7(d). 70
Figure 3.27. Amperometric responses of a biosensor to successive addition of 2 mM glucose under several applied potential (E vs. Ag/AgCl, counter=Pt plate). 72
Figure 3.28. Responses of enzyme electrodes prepared from different concentration of enzyme solution. 73
Figure 3.29. Responses to common physiological interferents. 74
Figure 3.30. CVs for electrochemical capping of microtubule array electrode after spin casting and membrane attachment procedure. The electrolyte solutions are 0.1 M LiClO₄/acetonitrile with (thick) and without... 75
Figure 3.31. Responses of the electrode prepared from different number of capping polymerization to common physiological interferents. Injection series and time are the same as in Figure 3.29.... 76
Figure 3.32. SEM images of the surface of microtubule electrode array after electrochemical capping. Monomer solution was 0.01 M 1,3-phenylenediamine/0.1 M LiClO₄/acetonitrile and the number of... 77
Figure 3.33. Amperometric responses of a capped biosensor to successive addition of 1 mM glucose. The inset is the calibration curve. 78
Figure 3.34. Lineweaver-Burke plot for getting apparent Michaelis constant. 79
Figure 3.35. Repeatability of the glucose sensor to 10 independent measurements. 5 mM glucose was injected at each run. 81
Figure 3.36. Response behavior of the glucose sensors with time elapse. They were stored at 4 ℃ in PBS and in air, respectively. The glucose concentration was 5 mM for each experiment. 82
Scheme 1.1. Generic electropolymerization pathway valid for many conducting polymers. 14
Scheme 2.1. Schematic diagram of the electrochemical synthesis of the hollow micro-tubule structure and capping process using conducting polymer composite. 31
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