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Cellular imaging techniques for neuroscience and beyond / [edited by] Floris G. Wouterlood. — London ; Waltham, MA : Elsevier/Academic Press, 2012. – (64.118/C393)

Contents

    CONTENTS
    
    List of Contributors
    Chapter 1 Confocal Laser Scanning: of Instrument, Computer Processing, and Men 1
     Introduction 2
     Pinhole, Depth of Focus, and Laser Illumination 2
     When/Why Does One Need a CLSM? 4
     Abbe, Shannon, and Nyquist 8
     Imaging of a 2D Line and Deblurring 9
     Axial Resolution 11
     Resolution and Sampling 12
     Signal Separation, Orders of Magnitude, and Resolution Limits 13
     Confocal Microscopy Further Considered 14
     Cross Talk Awareness 16
     Elimination of Cross Talk 18
     Biological Objects Translated to Pixels 19
     High-probability Determination of Diameter 20
     Why Does a 3D Reconstructed Cell Resemble a Pancake? 24
     Touch 25
     Actual Experiment 26
     Synaptic Contacts: Extra Marker 29
     Colocalization 30
     Conclusion 32
     Acknowledgments 32
     References 33
    Chapter 2 Beyond Abbe's Resolution Barrier: STED Microscopy 35
     Introduction 36
     A New Wave of Imaging 37
     STED Microscopy: The Basic Concept 39
    Implementation of STED Microscopy
    Sine Qua Non: Speed, Color, Depth, Live Imaging
    Summary and Outlook
    References
    Chapter 3 Enhancement of Optical Resolution by 4pi Single and Multiphoton Confocal Fluorescence Microscopy 55
     Introduction 56
     The 4pi Principle and Setup 56
     Microscope Alignment 58
     4pi Imaging 60
     4pi Deconvolution 61
     Sample Preparation 62
     Microtubule and Microtubule Plus End Imaging 65
     Visualization of DNA 66
     Single-photon Excitation (Measurement of the Redox State in Dopamine Neurons) 68
     SYCP3 Axis as a Marker for Chromatin Organization in Mouse Spermatocytes 70
     Microbubbles with Medicine 75
     Future of 4pi Imaging 77
     Acknowledgment 77
     References 77
    Chapter 4 Nano Resolution Optical Imaging Through Localization Microscopy 81
     Introduction 82
     Superresolution Microscopy Techniques 82
     The Main Approaches to Single-molecule Localization-based Superresolution Microscopy 87
     Fluorescent Probes 89
     Fluorescent Proteins 89
     Multicolor Localization Microscopy 90
    Outlook 93
    Conclusion 96
    Acknowledgments 96
    References 97
    Chapter 5 Optical Investigation of Brain Networks Using Structured Illumination 101
     Introduction 101
     Structuring Light by Phase Modulation Using SLMs 103
     Wavefront Engineering Using SLMs: The Optical Setup 104
     Light-sensitive Molecular Tools for the Investigation of the Central Nervous System 111
     SLM-based Approaches for the Optical Dissection of Brain Microcircuits 112
     Conclusions 116
     Acknowledgments 116
     References 116
    Chapter 6 Multiphoton Microscopy Advances Toward Super Resolution 121
     Introduction 121
     Point Spread Function for Single- and Multiphoton Imaging 123
     Super Resolution Techniques for Multiphoton Fluorescence Microscopy 126
     Conclusions 134
     Acknowledgments 134
     References 134
    Chapter 7 The Cell at Molecular Resolution: Principles and Applications of Cryo-Electron Tomography 141
     Introduction: Cellular Landscapes at Molecular Resolution 141
     The Cryo-ET Method 143
     Detection, Identification, and Hybrid Methods 151
    Conclusions 175
    Acknowledgments 178
    References 178
    Chapter 8 Cellular-Level Optical Biopsy Using Full-Field Optical Coherence Microscopy 185
     Introduction 185
     The FF-OCM Technique 187
     Detection Sensitivity 189
     Spatial Resolution 191
     Sample Motion Artifacts 192
     FF-OCM for High-Resolution "Optical Biopsy" 193
     Conclusion 195
     Acknowledgment 196
     References 196
    Chapter 9 Retroviral Labeling and Imaging of Newborn Neurons in the Adult Brain. 201
    Techniques to Label and Detect Newborn Neurons in the Adult Brain 202
    Retrovirus-mediated Labeling of Adult-born Neurons 203
    Single-cell Genetic Manipulation in Adult-born Neurons 205
    Retrovirus Production and Delivery 207
    Viral-labeled Cell Toxicity and Physiological Changes 209
    Imaging Newborn Neurons in the Adult Brain 210
    In Vivo Live Animal Imaging of Adult Neurogenesis 210
    In Vivo Window Preparation 211
    In Vivo Imaging Setup and Acquisition 213
    Postacquisition Image Processing and Analysis 213
    Future Directions in Live Animal Imaging of Adult Neurogenesis 214
    Acknowledgments 216
    References 216
    Chapter 10 Study of Myelin Sheaths by CARS Microscopy 221
     Traditional Myelin Imaging Methods 221
     Principle and History of CARS Microscopy 224
     Technical Characteristics of CARS Microscopy 226
     CARS Microscopy for Ex Vivo and In Vivo Myelin Imaging 227
     Mechanistic Understanding of Demyelination and Remyelination Enabled by CARS Imaging 231
     Other Methods for In Vivo Imaging of Myelin 236
     Outlook for Myelin Imaging by CARS Microscopy 238
     Acknowledgments 240
     References 240
    Chapter 11 High-Resolution Approaches to Studying Presynaptic Vesicle Dynamics Using Variants of FRAP and Electron Microscopy 247
     Introduction 248
     Quantifying Dynamic Events at the Macromolecular Scale 249
     FRAP for Studying Mobility 251
     Variations on FRAP Using Photoswitchable Fluorophores 255
     Linking Fluorescence and Ultrastructure: Correlative Approaches for Assaying Presynaptic Function 259
     Structure-Function Relationships of Vesicle Pools in Hippocampal Synapses 263
     Combining FRAP with Correlative Electron Microscope 265
     Concluding Remarks 269
     Acknowledgments 269
     References 270
    Index 275
    Color Plates