 |
Single cell analysis : technologies and applications / edited by Dario Anselmetti. — Weinheim : Wiley-VCH, c2009. – (58.15/S617) |
Contents
Contents
Foreword V
Preface XIII
List of Contributors XVII
Part I Single Cell Analysis : Imaging
1 Single Molecule Fluorescence Monitoring in Eukaryotic Cells: Intranuclear Dynamics of Splicing Factors 1
1.1 Motivation 1
1.2 Experimental Approach 2
1.3 Single Particle Tracking within Living Cells 6
1.4 Pre-Messenger RNA Splicing 7
1.5 Intranuclear Splicing Factor Tracking 8
1.6 Intranuclear U1 snRNP Splicing Factor Binding 10
1.7 Events in Speckles 10
1.8 Intranuclear U1 snRNP Mobility 11
1.9 Perspectives of Single Molecule Microscopy 13
References 15
2 Gene Classification and Quantitative Analysis of Gene Regulation in Bacteria using Single Cell Atomic Force Microscopy and Single Molecule Force Spectroscopy 19
2.1 Introduction 19
2.2 AFM on Paracrystalline Cell Surface Layers of C. glutarnicurn: Protein Sequence Information and Morphology 20
2.3 Imaging of Living C. glutamicum Cells with Molecular Resolution: Genes， Transcriptional Regulation and Morphology 23
2.4 Single Molecule Force Spectroscopy on Specific Protein-DNA Complexes: Transcriptional Regulation in S. rneliloti 25
2.5 Effector-Induced Protein-DNA Binding on the Single Molecule Level Quorum Sensing in S. meliloti 29
2.6 Conclusion 32
References 33
3 Cellular Cryo-Electron Tomography (CET): Towards a voyage to the inner space of cells Inner Space of Cells
3.1 Introduction 39
3.2 Tomography with the
3.3 Molecular Interpretation of Cellular Tomograms 58
3.4 Outlook: The Future is Bright 61
References 65
Part II Single Cell Analysis: Technologies
4 Single Cell Proteomics 69
4.1 Introduction 69
4.2 The Challenge 70
4.3 Single Cell Proteomics: Mass Spectrometry 71
4.4 Single Cell Separations 72
4.5 Ultrasensitive Protein Analysis: Capillary Electrophoresis with Laser-Induced Fluorescence Detection 74
4.6 Capillary Sieving Electrophoresis of Proteins from a Single Cancer Cell 75
4.7 Cell Cycle-dependent Single Cell Capillary Sieving Electrophoresis 77
4.8 Tentative Identification of Proteins in Single Cell Electropherograms 78
4.9 Capillary Micellar and Submicellar Separation of Proteins from a Single Cell 79
4.10 Two-Dimensional Capillary Electrophoresis of Proteins in a Single Cell 80
4.11 Single Copy Detection of Specific Proteins in Single Cells 83
4.12 Conclusion 85
References 87
5 Protein Analysis of Single Cells in Microfluidic Format 91
5.1 Introduction 91
5.2 Microfluidic Single Cell Analysis Concept 93
5.3 Single Cell Electrophoretic Separation and Detection of Proteins
5.4 Future Directions in Single Cell Analysis 103
References 104
6 Single Cell Mass Spectrometry 109
6.1 Introduction 109
6.2 Considerations for Single Cell Chemical Microanalysis using Mass Spectrometry 110
6.3 Mass Spectrometry as a Discovery Tool for Chemical Analysis of Cells 111
6.4 Single Cell Mass Spectrometric Applications 115
6.5 Subcellular Profiling 119
6.6 Imaging Single Cells with MS 121
6.7 Signaling Molecule Release from Single Cells 124
6.8 Future Developments 126
References 126
7 Single Cell Analysis for Quantitative Systems Biology 135
7.1 Introduction 135
7.2 Misleading Bulk Experiments 138
7.3 Common Techniques for High-Throughput and High-Content Single Cell Analysis 140
7.4 Improved Functionality for High-Throughput Single Cell Analysis 141
7.5 Example Studies Enabled by Microfluidic Cell Arrays 148
7.6 Conclusions and Future Directions 157
References 158
8 Optical Stretcher for Single Cells 161
8.1 Introduction 161
8.2 Theory， Methods and Experimental Setup 163
8.3 Applications 170
8.4 Outlook 173
References 174
Part III Single Cell Analysis: Applications
9 Single Cell Immunology 175
9.1 Introduction 175
9.2 Single Cell Gene Expression Profiling 175
9.3 Fluorescence-Activated Cell Sorting 180
9.4 Live Cell Fluorescence Microscopy 183
9.5 Other Techniques for Single Cell Analysis 187
9.6 Conclusions and Outlook 189
References 189
10 Molecular Characterization of Rare Single Tumor Cells 197
10.1 Introduction 197
10.2 Finding Rare Event Tumor Cells in Multidimensional Data 199
10.3 Classification of Rare Tumor Cells 205
10.4 Molecular Characterization of Sorted Tumor Cell Cells 209
10.5 Detection of Mutated Sequences in Tumor Suppressor Genes 211
10.6 Conclusions and Discussion 219
References 219
11 Single Cell Heterogeneity 223
11.1 Introduction 223
11.2 Measuring Heterogeneity using Single Cell Techniques 224
11.3 Describing Cellular Heterogeneities and Subpopulations 227
11.4 Origins of Cellular Heterogeneity 228
11.5 Identifying Extrinsic and Intrinsic Noise Sources 230
11.6 Concluding Remarks 233
References 234
12 Genome and Transcriptome Analysis of Single Tumor Cells 235
12.1 Introduction 235
12.2 Detection and Malignant Origin of Disseminated Cancer Cells 235
12.3 Methods for Amplifying Genomic DNA of Single Cells 237
12.4 Studying the Genome of Single Disseminated Cancer Cells 239
12.5 The Need for Higher Resolution: Array CGH of Single Cells 240
12.6 Studying the Gene Expression of Single Disseminated Cancer Cells 241
12.7 Combined Genome and Transcriptome Analysis of Single Disseminated Cancer Cells 246
References 246
Index 251
