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xi | |
Foreword |
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xiii | |
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1 Digital Microscopy: Nature to Numbers |
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1 | (30) |
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4 | (7) |
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1.1.1 First Principles: How Can Images Be Quantified? |
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4 | (2) |
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1.1.2 Representing Images as a Numerical Matrix Using a Scientific Camera |
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6 | (2) |
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1.1.3 Controlling Pixel Size in Cameras |
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8 | (3) |
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11 | (10) |
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12 | (1) |
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12 | (1) |
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12 | (2) |
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14 | (2) |
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16 | (3) |
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1.2.6 Registration and Calibration |
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19 | (2) |
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21 | (2) |
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23 | (6) |
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29 | (2) |
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2 Quantification of Image Data |
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31 | (16) |
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2.1 Making Sense of Images |
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31 | (4) |
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31 | (2) |
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2.1.2 Quantification of Image Data Via Computers |
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33 | (2) |
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2.2 Quantifiable Information |
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35 | (10) |
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2.2.1 Measuring and Comparing Intensities |
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35 | (1) |
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36 | (5) |
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2.2.3 Spatial Arrangement of Objects |
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41 | (4) |
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45 | (1) |
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46 | (1) |
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3 Segmentation in Bioimaging |
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47 | (36) |
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3.1 Segmentation and Information Condensation |
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47 | (5) |
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48 | (1) |
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3.1.2 An Intuitive Approach |
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49 | (2) |
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3.1.3 A Strategic Approach |
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51 | (1) |
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52 | (22) |
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3.2.1 Detecting and Counting Objects |
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52 | (8) |
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3.2.2 Automated Segmentation of Objects |
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60 | (14) |
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74 | (5) |
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79 | (4) |
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4 Measuring Molecular Dynamics and Interactions by Forster Resonance Energy Transfer (FRET) |
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83 | (16) |
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4.1 FRET-Based Techniques |
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83 | (6) |
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4.1.1 Ratiometric Imaging |
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84 | (1) |
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4.1.2 Acceptor Photobleaching |
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85 | (1) |
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4.1.3 Other FRET Measurement Techniques |
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85 | (2) |
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4.1.4 Alternative Methods to Measure Interactions |
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87 | (2) |
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89 | (3) |
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4.2.1 Ratiometric Imaging of FRET-Based Sensors |
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90 | (1) |
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4.2.2 Acceptor Photobleaching |
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91 | (1) |
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92 | (2) |
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4.3.1 Ratiometric Imaging |
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92 | (1) |
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4.3.2 Acceptor Photobleaching |
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93 | (1) |
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4.3.3 Data Averaging and Statistical Analysis |
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93 | (1) |
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4.4 Computational Aspects of Data Processing |
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94 | (1) |
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94 | (1) |
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4.4.2 FRET Data Analysis with Fiji |
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94 | (1) |
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95 | (1) |
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96 | (3) |
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5 FRAP and Other Photoperturbation Techniques |
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99 | (44) |
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5.1 Photoperturbation Techniques in Cell Biology |
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99 | (7) |
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5.1.1 Scientific Principles Underpinning FRAP |
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100 | (3) |
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5.1.2 Other Photoperturbation Techniques |
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103 | (3) |
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106 | (3) |
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5.2.1 Selecting Fluorescent Tags |
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107 | (1) |
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5.2.2 Optimisation of FRAP Experiments |
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107 | (2) |
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5.2.3 Storage of Experimental Data |
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109 | (1) |
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109 | (18) |
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5.3.1 Quantification of FRAP Intensities |
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112 | (1) |
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113 | (2) |
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5.3.3 In Silico Modelling of FRAP Data |
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115 | (5) |
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5.3.4 Fitting Recovery Curves |
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120 | (1) |
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5.3.5 Evaluating the Quality of FRAP Data and Analysis Results |
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121 | (1) |
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5.3.6 Data Averaging and Statistical Analysis |
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122 | (1) |
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5.3.7 Software for FRAP Data Processing |
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123 | (4) |
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5.4 Procedures for Quantitative FRAP Analysis with Freeware Software Tools |
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127 | (3) |
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5.4.1 Quantification of Intensity Traces with Fiji |
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127 | (1) |
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5.4.2 Processing FRAP Recovery Curves with FRAPAnalyser |
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128 | (2) |
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130 | (1) |
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131 | (1) |
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132 | (11) |
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5A Case Study: Analysing COPII Turnover During ER Exit |
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135 | (1) |
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5A.1 Quantitative FRAP Analysis of ER-Exit Sites |
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135 | (3) |
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5A.2 Mechanistic Insight into COPII Coat Kinetics with FRAP |
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138 | (2) |
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5A.3 Automated FRAP at ERESs |
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140 | (1) |
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141 | (2) |
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6 Co-Localisation and Correlation in Fluorescence Microscopy Data |
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143 | (30) |
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143 | (2) |
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6.2 Co-Localisation for Conventional Microscopy Images |
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145 | (19) |
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6.2.1 Co-Localisation in Super-Resolution Localisation Microscopy |
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151 | (5) |
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6.2.2 Fluorescence Correlation Spectroscopy |
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156 | (5) |
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6.2.3 Image Correlation Spectroscopy |
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161 | (3) |
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164 | (1) |
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165 | (1) |
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165 | (8) |
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7 Live Cell Imaging: Tracking Cell Movement |
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173 | (28) |
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173 | (1) |
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7.2 Setting up a Movie for Time-Lapse Imaging |
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174 | (1) |
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7.3 Overview of Automated and Manual Cell Tracking Software |
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175 | (9) |
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176 | (4) |
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180 | (1) |
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7.3.3 Comparison Between Automated and Manual Tracking |
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181 | (3) |
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7.4 Instructions for Using ImageJ Tracking |
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184 | (5) |
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7.5 Post-Tracking Analysis Using the Dunn Mathematica Software |
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189 | (9) |
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7.6 Summary and Future Direction |
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198 | (1) |
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198 | (3) |
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8 Super-Resolution Data Analysis |
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201 | (26) |
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8.1 Introduction to Super-Resolution Microscopy |
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201 | (1) |
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8.2 Processing Structured Illumination Microscopy Data |
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202 | (8) |
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8.2.1 SIM Reconstruction Theory |
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203 | (1) |
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8.2.2 Parameter Fitting and Corrections |
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204 | (1) |
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8.2.3 SIM Quality Control |
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205 | (1) |
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8.2.4 Checking System Calibration |
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205 | (1) |
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205 | (3) |
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8.2.6 Checking Reconstructed Data |
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208 | (1) |
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208 | (2) |
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8.3 Quantifying Single Molecule Localisation Microscopy Data |
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210 | (10) |
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8.3.1 SMLMS Pre-Processing |
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210 | (1) |
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8.3.2 Localisation: Finding Molecule Positions |
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210 | (1) |
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210 | (2) |
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8.3.4 Problem of Multiple Emissions Per Molecule |
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212 | (1) |
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8.3.5 Sieving and Quality Control and Drift Correction |
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213 | (5) |
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8.3.6 How Far Can I Trust the SMLM Data? |
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218 | (2) |
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8.4 Reconstruction Summary |
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220 | (1) |
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8.5 Image Analysis on Localisation Data |
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220 | (3) |
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221 | (1) |
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8.5.2 Stoichiometry and Counting |
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222 | (1) |
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8.5.3 Fitting and Particle Averaging |
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223 | (1) |
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223 | (1) |
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8.6 Summary and Available Tools |
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223 | (1) |
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224 | (3) |
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9 Big Data and Bio-Image Informatics: A Review of Software Technologies Available for Quantifying Large Datasets in Light-Microscopy |
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227 | (22) |
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227 | (1) |
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9.2 What Is Big Data Anyway? |
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228 | (3) |
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9.3 The Open-Source Bioimage Informatics Community |
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231 | (12) |
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9.3.1 ImageJ for Small-Scale Projects |
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231 | (4) |
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9.3.2 CellProfiler, Large-Scale Projects and the Need for Complex Infrastructure |
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235 | (3) |
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9.3.3 Technical Notes - Setting Up CellProfiler for Use on a Linux HPC |
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238 | (4) |
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9.3.4 Icy, Towards Reproducible Image Informatics |
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242 | (1) |
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9.4 Commercial Solutions for Bioimage Informatics |
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243 | (4) |
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243 | (1) |
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9.4.2 Definiens and Using Machine-Learning on Complex Datasets |
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244 | (3) |
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247 | (1) |
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247 | (1) |
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248 | (1) |
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10 Presenting and Storing Data for Publication |
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249 | (20) |
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10.1 How to Make Scientific Figures |
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249 | (7) |
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10.1.1 General Guidelines for Making Any Microscopy Figure |
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250 | (1) |
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10.1.2 Do's and Don'ts: Preparation of Figures for Publication |
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251 | (2) |
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10.1.3 Restoration, Revelation or Manipulation |
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253 | (3) |
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10.2 Presenting, Documenting and Storing Bioimage Data |
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256 | (11) |
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257 | (1) |
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10.2.2 The Open Microscopy Project |
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258 | (1) |
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10.2.3 OME and Bio-Formats, Supporting Interoperability in Bioimaging Data |
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259 | (1) |
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10.2.4 Long-Term Data Storage |
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260 | (2) |
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10.2.5 USB Drives Friend or Foe? |
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262 | (1) |
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10.2.6 Beyond the (USB) Drive Limit |
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262 | (1) |
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10.2.7 Servers and Storage Area Networks |
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263 | (2) |
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10.2.8 OMERO Scalable Data Management for Biologists |
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265 | (2) |
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267 | (1) |
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268 | (1) |
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11 Epilogue: A Framework for Bioimage Analysis |
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269 | (16) |
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11.1 Workflows for Bioimage Analysis |
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270 | (7) |
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270 | (2) |
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272 | (1) |
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11.1.3 Types of Workflows |
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273 | (3) |
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11.1.4 Types of Component |
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276 | (1) |
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11.2 Resources for Designing Workflows and Supporting Bioimage Analysis |
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277 | (5) |
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278 | (1) |
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11.2.2 A Network for Bioimage Analysis |
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279 | (1) |
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11.2.3 Additional Textbooks |
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279 | (1) |
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280 | (1) |
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11.2.5 Database of Components and Workflows |
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280 | (2) |
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11.2.6 Benchmarking Platform |
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282 | (1) |
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282 | (1) |
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283 | (2) |
Index |
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285 | |