Contributors |
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xiii | |
Preface |
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xix | |
Volumes in Series |
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xxi | |
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Section I Organelles and Cellular Structure |
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1 | (254) |
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1 Monitoring Cytoskeletal Dynamics in Living Neurons Using Fluorescence Photoactivation |
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3 | (20) |
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4 | (1) |
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2 Generation and Expression of PAGFP Fusion Constructs with Cytoskeletal Proteins |
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5 | (7) |
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3 Fluorescence Photoactivation, Live Cell Imaging, and Data Analysis |
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12 | (11) |
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20 | (1) |
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20 | (3) |
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2 Red/Far-Red Fluorescing DNA-Specific Anthraquinones for Nucl: Cyto Segmentation and Viability Reporting in Cell-Based Assays |
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23 | (24) |
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24 | (1) |
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2 The Basic Requirement for Counterstaining |
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25 | (2) |
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3 An Optimized Counterstain for GFP-Based Assays |
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27 | (1) |
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4 Cellular Compartment Segmentation |
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28 | (1) |
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5 Labeling Cells in Live and Fixed End-Point Assays |
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29 | (2) |
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6 Protocol for Differential Dual Nuclear/Cytoplasmic Counterstaining in Live Cells |
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31 | (1) |
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7 Protocols for Differential Dual Nuclear/Cytoplasmic Counterstaining in Fixed Cells |
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32 | (1) |
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8 Measuring Cellular DNA Content |
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33 | (1) |
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9 Protocol: DRAQ5™ DNA Content Measurement by Imaging |
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34 | (2) |
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10 Protocol: Cell Enumeration in "In-Cell Westerns" (After Hannoush, 2008) |
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36 | (1) |
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11 Further Augmentation of Compartment Segmentation |
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37 | (1) |
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12 Cell Enumeration and Morphometrics: Early Sentinels of In Vitro Toxicology |
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37 | (2) |
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13 A Far-Red Viability Dye Derivative |
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39 | (1) |
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14 Protocols: Use of DRAQ7™ to Identify Membrane-Compromised Cells |
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40 | (2) |
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42 | (5) |
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43 | (1) |
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43 | (4) |
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3 Methods to Measure Actin Treadmilling Rate in Dendritic Spines |
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47 | (12) |
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48 | (3) |
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51 | (8) |
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57 | (1) |
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57 | (2) |
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4 Live-Cell Imaging of Clathrin Coats |
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59 | (22) |
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60 | (1) |
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2 Dynamics of Clathrin Assembly |
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61 | (1) |
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3 Limits of Fluorescence Microscopy |
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62 | (1) |
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63 | (1) |
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5 How to Optimize Your TIRF System |
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64 | (1) |
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6 Spinning-Disk Confocal Microscopy |
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65 | (1) |
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7 Spherical Aberration Correction Applied to Spinning-Disk Confocal Microscopy |
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66 | (1) |
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8 Getting Around the Diffraction Limit |
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67 | (2) |
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9 Use of 2D Spinning-Disk Confocal Microscope to Study Clathrin-Mediated Endocytosis at the Ventral and Dorsal Surfaces of a Cell |
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69 | (1) |
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10 Use of TIRF Microscopy to Study Clathrin-Mediated Endocytosis |
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70 | (1) |
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71 | (1) |
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12 3D Tracking in Spinning-Disk Imaging |
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72 | (2) |
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13 Use of 3D Tracking to Monitor Clathrin-Mediated Entry of Reovirus Particles at the Apical Surface of Polarized Cells |
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74 | (2) |
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14 Using the Optimum Pixel and Step Sizes in z-Stacks |
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76 | (1) |
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76 | (5) |
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77 | (1) |
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77 | (4) |
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5 Imaging the Mitotic Spindle |
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81 | (24) |
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82 | (1) |
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2 Modern Light Microscopy |
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83 | (10) |
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93 | (8) |
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101 | (4) |
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101 | (1) |
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101 | (4) |
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6 Modeling of Spatial Intracellular Signaling Events in Neurons |
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105 | (20) |
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105 | (1) |
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2 Computational Modeling of Intracellular Signaling |
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106 | (6) |
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3 Imaging Signaling Events in Striatal Neuronal Cultures |
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112 | (9) |
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4 Experimental Validation of Model Predictions |
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121 | (4) |
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121 | (1) |
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121 | (4) |
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7 Imaging Chromosome Dynamics in Meiosis in Plants |
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125 | (20) |
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126 | (2) |
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2 Technical Considerations |
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128 | (6) |
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134 | (11) |
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141 | (1) |
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141 | (4) |
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8 Analyzing Lysosomes in Live Cells |
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145 | (14) |
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145 | (1) |
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146 | (6) |
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3 Measuring Lysosomal Ions |
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152 | (1) |
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4 Delivery of Phagocytic Material to the Lysosome |
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153 | (1) |
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5 Endocytosis of Membrane Probes to the Lysosome |
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154 | (5) |
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156 | (3) |
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9 Detection and Quantification of Biomolecular Association in Living Cells using Single-Molecule Microscopy |
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159 | (28) |
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1 Movements and Interactions Drive Biomolecular Function |
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160 | (2) |
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2 The Concept: "Please Move to the Dark, so That We Can See You" |
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162 | (2) |
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164 | (2) |
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166 | (10) |
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176 | (3) |
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179 | (2) |
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181 | (6) |
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182 | (1) |
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182 | (5) |
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10 Visualizing Cardiac Ion Channel Trafficking Pathways |
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187 | (16) |
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188 | (2) |
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2 Introducing Recombinant DNA for Ectopic Expression of Fluorescent Ion Channel Fusion Proteins |
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190 | (1) |
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3 Isolating De Novo Ion Channel Forward Trafficking |
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191 | (1) |
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4 Recreating Cardiomyocyte-Like Membrane Subdomains in Model Cell Systems |
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192 | (5) |
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5 Tools for Visualizing Forward Trafficking and Delivery of Ion Channels In Real Time |
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197 | (4) |
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201 | (1) |
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201 | (2) |
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202 | (1) |
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202 | (1) |
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11 Live Cell Imaging of the Cytoskeleton |
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203 | (16) |
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204 | (2) |
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2 Live Imaging of MTs In Tissue Cultured Cells |
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206 | (3) |
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3 Live Imaging of MTs in Whole Organisms |
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209 | (2) |
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4 Live Imaging of Actin Filaments in C. elegans |
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211 | (8) |
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214 | (5) |
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12 Fluorescence Single-Molecule Imaging of Actin Turnover and Regulatory Mechanisms |
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219 | (14) |
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220 | (2) |
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2 Introducing a Low Density of Fluorescent Proteins in XTC Cells |
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222 | (2) |
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3 Observation of XTC Cells Spreading on Poly-L-lysine (PLL)-coated Glass Coverslips |
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224 | (5) |
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229 | (4) |
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232 | (1) |
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232 | (1) |
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13 Analysis of Cell Dispersion and Migration by Video-Microscopy |
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233 | (22) |
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234 | (1) |
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235 | (1) |
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3 Model of Cell Dispersion |
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236 | (2) |
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4 Models of Cell Migration in Sheet |
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238 | (5) |
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5 Model of 3D Cell Migration |
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243 | (2) |
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245 | (7) |
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252 | (3) |
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252 | (3) |
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Section II Molecules and Ions |
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255 | (230) |
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14 Real Time Qualitative and Quantitative GLUT4 Translocation Assay |
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257 | (16) |
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Maleppillil Vavachan Vijayakumar |
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258 | (1) |
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2 GLUT4 Translocation Assay |
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259 | (2) |
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261 | (9) |
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4 Conclusions and Future Applications |
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270 | (3) |
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270 | (1) |
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270 | (3) |
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15 Advanced Imaging of Cellular Signaling Events |
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273 | (18) |
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274 | (1) |
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2 Selection of Tools and Transfection of Cells |
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275 | (1) |
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3 Imaging of Cell Signaling Events |
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276 | (5) |
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281 | (6) |
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287 | (4) |
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288 | (1) |
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288 | (3) |
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16 Dynamic Imaging of Homo-FRET in Live Cells by Fluorescence Anisotropy Microscopy |
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291 | (38) |
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292 | (1) |
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293 | (5) |
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3 Homo-FRET Measurements in an Imaging Modality: Practical Considerations |
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298 | (11) |
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4 Implementation of High-Resolution Steady-State Anisotropy Imaging Modalities |
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309 | (10) |
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5 Detecting Homo-FRET Using Time Resolved Anisotropy (TRA) Modalities |
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319 | (6) |
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6 Prospects and Perspectives |
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325 | (4) |
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325 | (1) |
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325 | (4) |
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17 Time-Resolved Luminescence Resonance Energy Transfer Imaging of Protein-Protein Interactions in Living Cells |
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329 | (18) |
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330 | (2) |
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2 Lanthanide Protein Labels |
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332 | (3) |
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3 Transfection and Cellular Delivery of TMP-Lumi4 |
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335 | (3) |
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4 Time-Resolved LRET Microscopy |
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338 | (9) |
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344 | (1) |
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344 | (3) |
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18 Imaging of Corticosteroid Receptors in Live Cells |
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347 | (16) |
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348 | (1) |
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2 Time-Lapse Imaging of GR and MR in a Single Cell |
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349 | (5) |
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3 Subcellular and Intranuclear Dynamics of GR and MR Examined by FRAP Analyses |
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354 | (1) |
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4 Interaction of Corticosteroid Receptors in the Nucleus |
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354 | (3) |
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357 | (6) |
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360 | (1) |
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360 | (3) |
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19 Investigating Second Messenger Signaling In Vivo |
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363 | (20) |
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364 | (1) |
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2 Multimodal Imaging in Living Mouse Skeletal Muscle |
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365 | (10) |
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3 Adaptations to Tissues Other Than Skeletal Muscle |
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375 | (5) |
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380 | (3) |
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380 | (1) |
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380 | (3) |
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20 Probes for Intracellular RNA Imaging in Live Cells |
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383 | (18) |
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384 | (1) |
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2 Imaging RNA in Live Cells |
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385 | (1) |
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3 Limitations of Single-Label Probes and Molecular Beacons |
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386 | (3) |
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4 Imaging Native, Nonengineered RNA in Live Cells with Single RNA Sensitivity Using Exogenous Probes |
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389 | (5) |
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5 Time-Lapse Imaging of Native, Nonengineered β-actin mRNA Granule Dynamics |
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394 | (1) |
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6 Imaging RNA-Protein Colocalization Using MTRIPs |
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395 | (1) |
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396 | (5) |
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397 | (4) |
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21 Imaging the Glycome in Living Systems |
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401 | (20) |
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402 | (1) |
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2 Bioorthogonal Chemistry in Glycan Labeling: Merits and Limitations |
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403 | (3) |
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3 Metabolic Labeling of Cell Surface Fucosylated Glycans in Zebrafish Embryos for Fluorescence Imaging |
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406 | (6) |
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4 Labeling Cell Surface Glycans Bearing the LacNAc Disaccharide in Chinese Hamster Ovary(CHO) Cells for Fluorescence Imaging |
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412 | (9) |
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416 | (1) |
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416 | (5) |
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22 Intracellular Magnesium Detection by Fluorescent Indicators |
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421 | (24) |
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422 | (2) |
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2 Fluorescent Indicators for Magnesium |
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424 | (4) |
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3 Magnesium Detection in Live Cells by Spectrofluorimetry |
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428 | (6) |
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4 Magnesium Detection in Live Cells by Microscopy |
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434 | (7) |
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441 | (4) |
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442 | (1) |
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442 | (3) |
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23 Illuminating Mobile Zinc with Fluorescence: From Cuvettes to Live Cells and Tissues |
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445 | (24) |
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446 | (2) |
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2 A Guide to Fluorescent Zinc Probes |
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448 | (9) |
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3 Representative Procedure for Imaging Mobile Zinc in Live Cells |
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457 | (5) |
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4 Imaging Endogenous Neuronal Zinc in Live Cells and Tissues |
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462 | (2) |
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464 | (5) |
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465 | (1) |
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465 | (4) |
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24 Quantitative Fluorescent Live Cell Imaging of Intracellular Ca2+ and H+ Ions in Malaria Parasites |
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469 | (16) |
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470 | (1) |
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2 Live Cell Imaging of Intact Malaria Parasites |
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471 | (3) |
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3 Measuring Ion Homeostasis in P. falciparum Using Fluorescent Indicators |
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474 | (11) |
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482 | (1) |
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482 | (3) |
Author Index |
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485 | (26) |
Subject Index |
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511 | |