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Chemical cytometry : ultrasensitive analysis of single cells / edited by Chang Lu. — Weinheim : Wiley-VCH, 2010. – (58.1573/C517) |
Contents
Contents
Preface
List of Contributors
1 Origin， Current Status， and Future Perspectives of Chemical Cytometry 1
1.1 The Cell and Cytometry 1
1.2 Flow and Image Cytometry 1
1.3 Chemical Cytometry 2
1.4 Chemical Cytometry of DNA and mRNA 10
1.5 Metabolic Cytometry 10
1.6 Future Perspectives on Instrumentation for Chemical Cytometry 12
Acknowledgments 14
References 14
2 Metabolic Cytometry - The Study of Glycosphingolipid Metabolism in Single Primary Cells of the Dorsal Root Ganglia 21
2.1 Introduction 21
2.2 Material and Methods 24
2.3 Results and Discussion 26
2.4 Conclusions 29
Acknowledgments 29
References 29
3 Cell Signaling Studied at the Single-cell Level 31
3.1 Introduction 31
3.2 Analytes Examined and Reporters Used 32
3.3 Cell Preparation and Reporter Loading 35
3.4 Cell Lysis and Sampling Techniques 37
3.5 Electrophoresis Separation Conditions 43
3.6 Detection 46
3.7 Automation and Throughput 50
References 52
4 Ultrasensitive Detection of Low-copy-number Molecules from Single Cells 55
4.1 Introduction 55
4.2 Microchip Designs for Single-cell Analysis and/or Cell Manipulation 55
4.3 Ultrasensitive Detection Methods for Single-cell Analysis 57
4.4 Single-cell Analysis with Single-molecule Sensitivity on Integrated Microfluidic Chip 65
4.5 Conclusions 71
References 71
5 Capillary Electrophoresis of Nucleic Acids at the Single-cell Level
5.1 Introduction 75
5.2 On-line Cell Analysis 76
5.3 Direct Gene and Gene Expression Analysis Without Amplification
5.4 Potential Alternative Techniques for Single-cell Gene and Gene Expression Analysis 88
5.5 Conclusions 90
References 90
6 Microfluidic Technology for Single-cell Analysis 93
6.1 Introduction 93
6.2 Biological Significance of Single-cell Analysis 94
6.3 Microfluidic Devices in Our Laboratories 96
6.4 Materials， Methods， and Protocols 100
6.5 Conclusions 104
References 105
7 On-chip Electroporation and Electrofusion for Single-cell Engineering 107
7.1 Introduction 107
7.2 Single-cell Electroporation in Microfluidic Devices 108
7.3 Single-cell Electrofusion in Microfluidic Devices 117
7.4 Conclusions 120
References 121
8 Electroporative Flow Cytometry for Single-cell Analysis 125
8.1 Introduction 125
8.2 Flow-through Electroporation under Constant Voltage 126
8.3 Electroporative Flow Cytometry for Detecting Protein Translocation 128
8.4 Electroporative Flow Cytometry for Measuring Single-cell Biomechanics 134
8.5 Electroporative Flow Cytometry for Selectively Releasing and Analyzing Specific Intracellular Molecules 137
8.6 Conclusions 139
Acknowledgments 139
References 139
9 Ultrasensitive Analysis of Individual Cells via Droplet Microfluidics 143
9.1 Introduction 143
9.2 Droplet Properties 143
9.3 Droplet Generation 144
9.4 Cell Encapsulation 146
9.5 Droplet Manipulation 147
9.6 Droplet Concentration Control 149
9.7 Temperature Control of Droplets 150
9.8 Detection in Droplets 152
9.9 Conclusions 155
References 155
10 Probing Exocytosis at Single Cells Using Electrochemistry 159
10.1 Introduction 159
10.2 Measurement Requirements 160
10.3 Electrode Fabrication 160
10.4 Measurements at Single Cells 161
10.5 Fusion Pore Dynamics 163
10.6 Conclusions 170
Acknowledgments 171
References 171
11 Electrochemical Determination of Enzyme Activity in Single Cells 175
11.1 Introduction 175
11.2 Electrochemical Detection Coupled with Capillary Electrophoresis 175
11.3 Voltammetry 180
11.4 Scanning Electrochemical Microscopy (SECM) 183
11.5 High-throughput ECD 188
11.6 Perspective 190
Acknowledgment 190
References 191
12 Single-cell Mass Spectrometry 197
12.1 Introduction 197
12.2 Mass Spectrometry 198
12.3 Overall Outlook for Single-cell MS 213
Acknowledgments 213
References 213
13 Optical Sensing Arrays for Single-cell Analysis 219
13.1 Introduction to Fiber-optic Single-cell Arrays 219
13.2 Advantages of Fiber-optic Single-cell Arrays 220
13.3 Fiber-optic Arrays 222
13.4 Single-cell Arrays for Bacteria 223
13.5 Single-cell Arrays for Yeast 226
13.6 Single-cell Arrays for Mammalian Cells 229
13.7 Image and Data Analysis for Single-cell Arrays 234
13.8 Summary 234
References 236
Index 239
