Title Page
Contents
ABSTRACT IN KOREAN 17
ABSTRACT 20
PART I. General Introduction 23
I. SINGLE-ENTITY ELECTROCHEMISTRY 23
1.1. Single-Entity Electrochemistry (SEE) 23
1.2. Electrochemical Method: Different Types of SEE Signals 24
II. REFERENCES 26
PART II. Janus Emulsion Detection Using Characteristics of Interfaces Between Two or More Immiscible Electrolyte Solutions 27
I. INTRODUCTION 27
II. EXPERIMENTAL DETAILS 31
2.1. Reagents 31
2.2. Instrumentation 31
2.3. Preparation of Janus emulsion droplets 32
2.4. Phase change of Janus emulsion droplets 35
III. RESULTS AND DISCUSSION 37
3.1. Determination of Applied Potential of Ferrocene on an Ultramicroelectrode 37
3.2. w and w/o capstone i-t curves 42
3.3. Calculation of Janus Emulsion Concentration from Number of Collision 44
IV. SUMMARY AND CONCLUSIONS 47
V. REFERENCES 48
ABSTRACT 53
PART III. Serotonin in Single Human Platelets Detection Method 54
I. INTRODUCTION 54
II. EXPERIMENTAL DETAILS 59
2.1. Reagents and Materials 59
2.2. Instrumentation 59
2.3. Sample Preparation 60
2.4. The Fabrication of an Ultramicroelectrode (UME) 60
2.5. Calculating Concentration of Serotonin in a Single Platelet using a Chronoamperometry 61
III. RESULTS AND DISCUSSION 63
3.1. Consideration of Serotonin Reaction Potential using a Bulk Electrode 63
3.2. Observation of Serotonin Oxidation Signal in a Single Human Platelet on a Pt UME 66
3.3. Validation of Serotonin Concentration in a Single Human Platelet 72
3.4. Interpretation of Collision Signal Considering S/N ratio and Analysis of Serotonin Concentration through the Assumption 75
3.5. Determination of Platelet Concentration through Collision Frequency 77
IV. SUMMARYAND CONCLUSIONS 80
V. REFERENCES 81
ABSTRACT 86
PART IV. Electric Field Active Probe: Electrophoretic Migration Mediated Sensor for Sensitive Detection of Bio-entity 87
I. INTRODUCTION 87
1.1. Electrophoretic Migration of Mass Transfer 87
1.2. Electric Fields Active Probe (EP) 90
1.3. General Introduction 92
II. EXPERIMENTAL DETAILS 96
2.1. Reagents 96
2.2. Instrumentation 97
2.3. Antibody-DNA Conjugation for Electric Field Active Probe (EP) 97
2.4. Measurement of MNCs with EPs using the Current-Time (i-t) Method 100
2.5. Measurement of MNCs with EPs using the Current-Time (i-t) Method 100
2.6. COMSOL Multiphysics® 5.6[이미지참조] 103
III. RESULTS AND DISCUSSION 104
3.1. Antibody-DNA Conjugation of EP 104
3.2. Validation of Collision Signal Amplification by the EP 107
3.3. Determination of Buffer pH and Ferrocyanide Oxidation Potential of Solutions for Electrochemical Collision of MNCs with EP 109
3.4. Determination of the Ratio of MNCs to EP for Collision Signal 114
3.5. Detection of MNCs with EP and Determination of Frequency of Collision 116
3.6. Effect of MNC with EP using Simulated Current Decrease 122
IV. SUMMARY AND CONCLUSIONS 126
V. REFERENCES 127
ABSTRACT 132
Table 2.1. Diameter distribution of individual Janus emulsion obtained by... 46
Table 4.1. COMSOL Simulation result of MNCs collision in 3D space. 124
Table 4.2. Experimental result of MNCs collision. 125
Figure 1.1. Schematic representation of each stochastic technique for single entity... 25
Figure 2.1. Schematic Illustration of the collision of Janus emulsion droplets.... 30
Figure 2.2. Schematic illustration of Janus emulsion. 34
Figure 2.3. Dynamic morphology changes of emulsion droplets. 36
Figure 2.4. Cyclic voltammograms of the with a 10 ㎛ C-UME with and without... 39
Figure 2.5. Cyclic voltammograms of the with a 10 ㎛ C-UME with 0.05, 0.1, 0.5... 40
Figure 2.6. i-t curves of Janus emulsion collision on a C UME at different applied... 41
Figure 2.7. i-t curves of Janus emulsion collision under various applied potential... 43
Figure 2.8. Size distribution values of the DLS method (black line) and integrated... 45
Figure 3.1. Schematic Diagram of Serotonin Oxidation Using SEE in a Single... 58
Figure 3.2. Complete blood count (CBC) obtained from the Advia2120i Auto... 62
Figure 3.3. (a) CVs of PPP (black line) and PRP (red line) with a 25 ㎛ Pt UME at... 65
Figure 3.4. i-t curves of (a) PPP, (b) 10.0, (c) 20.1, (d) 40.2, and (e) 100 fM... 68
Figure 3.5. i-t curves of PRP in RLB at Pt UME (25 ㎛ diameter) under applied... 69
Figure 3.6. i-t curves of PRP (100 fM of platelets, red line), PPP (black line), and... 70
Figure 3.7. i-t curves of (a) PPP, (b) 10.0, (c) 20.1, (d) 40.2, and (e) 100 fM... 71
Figure 3.8. (a) Integral area (red dashed line) of a single current spike because of... 74
Figure 3.9. An example of spike signal determination considering S/N ratio. 76
Figure 3.10. Regression plot of the collision frequency as a function of the number... 79
Figure 4.1. Schematic diagram of a system for measuring blood entities... 94
Figure 4.2. Schematic diagram of MNC-specific EP for a novel electrochemical... 95
Figure 4.3. Schematic diagram of antibody-DNA conjugation for the measurement... 99
Figure 4.4. White blood cell count obtained from the instrument. 102
Figure 4.5. DLS analysis of solutions (a) CD14 antibody (AB, blue line) and DNA... 106
Figure 4.6. i-t curves of without MNC and EP (black line), 83.9 aM MNC without... 108
Figure 4.7. i-t curves of 83.9 aM monocytes with EP (0.5 ㎍ EP per 106 cell MNC...[이미지참조] 111
Figure 4.8. i-t curves of monocytes with EP (1.0 ㎍ EP per 106 cell MNC ratio) in...[이미지참조] 112
Figure 4.9. CVs of TBS pH 9.0 solution (black line), MNC without EP (red line),... 113
Figure 4.10. i-t curves of 83.9 aM monocytes with (a) without EP, (b) 0.5, (c) 1.0,... 115
Figure 4.11. i-t curves of (a) 21.0, (b) 41.9, and (c) 83.9 aM monocytes with EP... 120
Figure 4.12. Regression plot of the collision frequency as a function of the number... 121
Figure 4.13. (a) COMSOL simulation for measuring the decrease in current when... 123