| Preface |
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xv | |
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1 A Short Story of AFM in Biology |
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1 | (12) |
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1.1 Development of Biological AFM |
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2 | (4) |
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1.1.1 From Contact Mode to Tapping Mode |
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3 | (1) |
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1.1.2 Technology Development toward High-Resolution Imaging |
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4 | (1) |
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5 | (1) |
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1.1.4 Force Measurement and Recognition Imaging |
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6 | (1) |
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1.2 Mapping Bio-AFM Research |
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6 | (7) |
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1.2.1 Blooming Activities of AFM Research Communities |
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6 | (2) |
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1.2.2 AFM: One of the Top Keywords in Biological Research |
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8 | (5) |
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2 Protocols for Specimen and Substrate Preparation and Data Correction Methods |
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13 | (20) |
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2.1 Atomic Force Microscopy |
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14 | (1) |
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15 | (3) |
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18 | (1) |
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2.4 Setup and Measurement |
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19 | (1) |
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20 | (2) |
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22 | (7) |
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23 | (1) |
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24 | (2) |
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26 | (3) |
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29 | (4) |
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3 Chemical Modification of AFM Probes and Coupling with Biomolecules |
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33 | (14) |
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3.1 Chemical Modification of AFM Probes and other Inorganic Materials |
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34 | (3) |
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3.2 Coupling Proteins to the Cantilever Surface |
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37 | (5) |
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37 | (2) |
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3.2.2 Site-Specific Attachment |
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39 | (3) |
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42 | (1) |
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3.4 Applications of Modified Cantilevers |
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42 | (2) |
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3.5 Conclusions and Outlook |
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44 | (3) |
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4 Single-Molecule Dissection and Isolation Based on AFM Nanomanipulation |
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47 | (14) |
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47 | (1) |
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4.2 AFM Dissection and Patterning of Individual DNA Molecules |
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48 | (2) |
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4.3 Isolation of DNA by AFM Nanomanipulation |
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50 | (3) |
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4.4 Positioning Scission of Single DNA with Nonspecific Endonuclease |
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53 | (1) |
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4.5 Defect Repair and Guided Growth of Peptide Nanofilaments |
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54 | (3) |
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57 | (4) |
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5 Structural Biology with Cryo-AFMs and Computational Modeling |
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61 | (24) |
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61 | (2) |
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63 | (3) |
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66 | (6) |
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5.4 Combining a Cryo-AFM with Molecular Modeling |
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72 | (6) |
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78 | (7) |
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6 High-Resolution Imaging of Biological Molecules by Frequency Modulation Atomic Force Microscopy |
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85 | (26) |
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85 | (1) |
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6.2 Instrumentation of FM-AFM |
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86 | (8) |
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6.2.1 Cantilever as a Force Sensor |
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86 | (2) |
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6.2.2 Detection of the Cantilever Resonance Frequency Shift |
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88 | (2) |
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6.2.3 Instrumentation of the FM Detection Method |
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90 | (2) |
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6.2.4 Conversion of Frequency Shift to Interaction Force |
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92 | (2) |
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6.3 Problems of FM-AFM in Liquids |
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94 | (4) |
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6.3.1 Viscous Damping of a Cantilever in Fluid |
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94 | (2) |
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6.3.2 Electric Double-Layer Force |
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96 | (2) |
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6.4 High-Resolution Imaging by FM-AFM in Liquids |
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98 | (2) |
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6.5 Purple Membrane Proteins |
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100 | (2) |
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6.6 Isolated Chaperonin Proteins |
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102 | (1) |
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6.7 Force Mapping Techniques Using FM-AFM in Liquids |
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103 | (5) |
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6.7.1 Visualization of Hydration Layers at the Mica--Water Interface |
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103 | (3) |
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6.7.2 Two-Dimensional Force Mapping at the Graphite--Water Interface |
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106 | (2) |
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108 | (3) |
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7 Development of Recognition Imaging: From Molecules to Cells |
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111 | (32) |
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111 | (2) |
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113 | (3) |
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7.3 The Working Principle of TREC |
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116 | (8) |
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7.3.1 Feedback, Working Amplitude, and Frequency Optimization |
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117 | (6) |
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7.3.2 Specificity Proof for the Detected Interactions |
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123 | (1) |
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7.4 Application 1: Single-Molecule TREC on Biotinylated DNA Tetrahedra |
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124 | (6) |
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124 | (1) |
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7.4.2 A Short Introduction to DNA Building Blocks |
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125 | (1) |
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7.4.3 Imaging of Single DNA Tetrahedra |
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126 | (2) |
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7.4.4 Single-Molecule TREC on Biotinylated DNA Tetrahedra |
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128 | (2) |
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130 | (1) |
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7.5 Application 2: Nanolandscape of FCγ Receptors on the Macrophage Surface |
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130 | (13) |
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130 | (1) |
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7.5.2 Phagocytosis and Phagocytic Receptors |
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131 | (1) |
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7.5.3 Binding Capacity of Fcγ Receptors on the Macrophage Surface |
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132 | (2) |
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7.5.4 Nanomapping of Fcγ Rs |
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134 | (4) |
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138 | (5) |
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8 Development of High-Speed AFM and Its Biological Applications |
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143 | (34) |
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144 | (1) |
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8.2 Factors Limiting Scan Speed |
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145 | (2) |
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147 | (12) |
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147 | (3) |
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150 | (1) |
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8.3.3 Fast Amplitude Detector |
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151 | (2) |
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153 | (4) |
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8.3.5 High-Speed and Low-Invasive Control Methods |
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157 | (2) |
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8.4 Substrate Surfaces for Dynamic AFM Imaging of Biomolecules in Action |
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159 | (5) |
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8.4.1 Mica-Supported Planar Lipid Bilayers |
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161 | (2) |
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8.4.2 2D Crystals of Streptavidin |
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163 | (1) |
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8.5 Biological Applications |
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164 | (13) |
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8.5.1 Walking Mechanism of Myosin V |
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164 | (5) |
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8.5.2 Photoinduced Conformational Change in Bacteriorhodopsin |
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169 | (8) |
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9 Real-Time AFMs Combined with Inverted Optical Microscopes for Wet Cell/Tissue Imaging |
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177 | (14) |
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178 | (6) |
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179 | (2) |
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9.1.2 Demonstration of the Functions of This Instrument |
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181 | (3) |
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9.2 Biological Application |
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184 | (3) |
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9.2.1 Live Cell Imaging in Solution |
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184 | (1) |
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9.2.2 Tissue Imaging in Solution |
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185 | (2) |
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187 | (4) |
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10 Studying the Cytoskeleton by Atomic Force Microscopy |
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191 | (28) |
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191 | (5) |
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10.1.1 The Cytoskeleton: A Complex Scaffold Determining Cell Shape and Mechanics |
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192 | (1) |
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192 | (1) |
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193 | (2) |
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10.1.4 Intermediate Filaments |
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195 | (1) |
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10.2 Maging the Cytoskeleton of Living Cells |
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196 | (7) |
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10.2.1 Immobilizing Cells for AFM Scanning |
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196 | (1) |
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10.2.2 Visualizing the Cortical Actin Cytoskeleton |
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196 | (2) |
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10.2.3 Time-Lapse Imaging of Living Cells |
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198 | (2) |
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10.2.4 Investigating Intracellular Compartments in De-Roofed Cells |
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200 | (3) |
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10.3 Imaging Cytoskeletal Filaments in vitro |
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203 | (10) |
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10.3.1 Imaging Actin Filaments in vitro |
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203 | (1) |
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10.3.2 Immobilizing Microtubules for AFM Scanning in vitro |
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204 | (2) |
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10.3.3 Imaging Chemically Fixed and Unfixed Microtubules |
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206 | (2) |
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10.3.4 Dynamic Microtubules |
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208 | (1) |
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10.3.5 Imaging Single Intermediate Filaments in vitro |
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208 | (2) |
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10.3.6 Investigating the Mechanical Behavior of Single Intermediate Filaments |
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210 | (3) |
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213 | (6) |
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11 Determination of the Architecture of Multisubunit Proteins Using AFM Imaging |
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219 | (26) |
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219 | (2) |
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221 | (2) |
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11.3 Antibody Decoration of Epitope Tags |
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223 | (1) |
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224 | (1) |
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224 | (4) |
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228 | (3) |
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11.7 The TRPP2/TRPC1 Channel |
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231 | (2) |
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11.8 The Epithelial Sodium Channel |
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233 | (4) |
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237 | (8) |
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12 Capturing Membrane Proteins at Work |
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245 | (14) |
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12.1 AFM for the Structural Analyses of Membrane Proteins |
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246 | (1) |
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12.2 Conformational Changes of Ligand-Gated Ion Channels |
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247 | (5) |
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12.2.1 Agonist-Induced Structural Changes in the NMDA Receptor |
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247 | (3) |
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12.2.2 ATP-Induced Conformational Changes in the P2X4 Receptor |
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250 | (2) |
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12.3 Direct Visualization of the Albers--Post Scheme of P-Type ATpases |
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252 | (4) |
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12.4 Conclusion and Perspectives |
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256 | (3) |
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13 Enzymes and DNA: Molecular Motors in Action |
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259 | (16) |
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13.1 Atomic Force Microscopy and DNA |
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259 | (2) |
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13.2 Background to Restriction-Modification Systems |
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261 | (14) |
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262 | (3) |
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265 | (3) |
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268 | (3) |
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271 | (4) |
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14 Genome-Folding Mechanisms in the Three Domains of Life Revealed by AFM |
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275 | (36) |
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14.1 Biophysical Properties of DNA and DNA-Binding Proteins |
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276 | (4) |
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14.1.1 Persistence Length and Phase Transition of DNA Conformation |
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276 | (1) |
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14.1.2 Principles of DNA-Protein Interaction |
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277 | (3) |
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14.2 Nucleosome and beyond in Eukaryotes Revealed by AFM |
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280 | (5) |
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14.2.1 Nucleosome Reconstitution |
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280 | (1) |
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14.2.2 Reconstitution of Higher-Order Structures of Chromatin |
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281 | (3) |
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14.2.3 Genome Architecture in vivo |
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284 | (1) |
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14.3 Evolutionary Aspects of Genome Architectures in Bacteria and Archaea |
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285 | (17) |
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14.3.1 Bacterial Nucleoid Architecture in vivo |
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287 | (3) |
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14.3.2 In vitro Reconstitution of the Bacterial Nucleoid |
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290 | (1) |
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14.3.3 In vivo Dynamics of the Bacterial Nucleoid |
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291 | (3) |
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14.3.4 Archaeal Chromosomal Proteins |
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294 | (1) |
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14.3.5 Archaeal Chromosome Architectures |
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295 | (4) |
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14.3.6 Archaeal Nucleoid Dynamics in vivo |
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299 | (3) |
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14.4 Conclusion and Perspectives |
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302 | (9) |
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15 Membrane Dynamics: Lipid--Protein Interactions Studied by AFM |
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311 | (18) |
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15.1 AFM as an Analytical Tool for the Study of Membrane Dynamics |
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312 | (1) |
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15.2 AFM Imaging of the Interaction between SNARE Proteins and Membranes |
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312 | (6) |
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312 | (1) |
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15.2.2 SNARE Proteins and the Lipid Bilayer |
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313 | (4) |
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15.2.3 Recognition ("Mapping") Imaging of Proteins Involved in Membrane Fusion |
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317 | (1) |
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15.3 Force Spectroscopy Addressing the Physical Mechanisms of Membrane Fusion |
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318 | (4) |
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15.3.1 Physical Properties of SNAREs and Synaptotagmin |
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318 | (2) |
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15.3.2 Physical Properties of the Synaptotagmin-Lipid Interaction |
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320 | (2) |
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322 | (1) |
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322 | (1) |
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323 | (1) |
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323 | (6) |
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16 Nanosurgery and Cytoskeletal Mechanics of a Single Cell |
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329 | (28) |
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Takahiro Watanabe Nakayama |
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16.1 Delivery/Extraction of Nucleic Acid from a Single Cell |
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330 | (6) |
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16.1.1 Retrieval of DNA from Chromosomes |
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330 | (2) |
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16.1.2 Retrieval of mRNA from Individual Cells |
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332 | (1) |
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16.1.3 Insertion of DNA into Individual Cells |
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332 | (4) |
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16.2 Manipulation of the Red Blood Cell Cytoskeleton |
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336 | (2) |
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16.3 Mechanics of Fibroblast Stress Fibers with a Lateral Force |
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338 | (10) |
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16.4 Manipulation of Stress Fibers by FIB-Fabricated Probes |
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348 | (3) |
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16.5 Hole Creation on the Cell Surface |
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351 | (3) |
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16.6 Conclusion and Perspectives: Cellular Mechanics Probed with AFM |
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354 | (3) |
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17 Functional Investigations on Nuclear Pores with Atomic Force Microscopy |
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357 | (22) |
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17.1 Atomic Force Microscopy |
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358 | (1) |
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17.2 AFM to Probe Biological Samples |
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359 | (15) |
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17.2.1 AFM-Based Functional Investigations on Nuclear Pore Complexes |
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361 | (1) |
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17.2.2 The AFM Tip as a Chemical Nanosensor to Explore the Hydrophobicity of Intact and Apoptotic NPC Channels |
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362 | (5) |
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17.2.3 The AFM Tip as a Nanoindentor to Study the Mechanical Properties of the NPC |
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367 | (1) |
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17.2.4 The Atomic Force Microscope Tip as a Surgical Nanotool to Harvest Transcripts of Early Genes from the NPC |
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368 | (6) |
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17.3 Conclusions and Outlook |
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374 | (5) |
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18 Mechanotransduction: Probing Its Mechanisms at the Nanoscale Using the Atomic Force Microscope |
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379 | (44) |
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18.1 Cellular Mechanotransduction |
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379 | (5) |
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18.2 Cellular Elasticity and What It Tells Us |
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384 | (7) |
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18.2.1 AFM Force--Distance Curves |
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384 | (6) |
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18.2.2 Material Properties of the Cell |
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390 | (1) |
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18.3 Cellular Force Transducers |
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391 | (23) |
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18.3.1 Cellular Deformation |
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395 | (1) |
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18.3.1.1 Whole-cell deformation |
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396 | (2) |
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398 | (2) |
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18.3.1.3 Localized deformation |
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400 | (3) |
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18.3.2 Cytoskeletal Dynamics |
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403 | (1) |
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403 | (4) |
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18.3.2.2 Mitochondrial dynamics |
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407 | (1) |
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18.3.2.3 Inducing traction forces |
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408 | (4) |
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18.3.3 Inducing a Visible Biochemical Response |
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412 | (1) |
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18.3.3.1 Cell--cell calcium signaling |
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412 | (2) |
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18.4 Conclusions and Outlook |
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414 | (9) |
| Index |
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