Methods in Enzymology; vol.504: Imaging and spectroscopic analysis of living cells: Optical and spectroscopic techniques (San Diego; London, 2012). - ОГЛАВЛЕНИЕ / CONTENTS
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ОбложкаMethods in Enzymology. Vol.504: Imaging and spectroscopic analysis of living cells: Optical and spectroscopic techniques. - San Diego; London: Academic Press; Elsevier, 2012. - xlviii, 506 p.: ill. - Incl. bibl. ref. and indexes. - ISBN 978-0-12-391857-4
 

Оглавление / Contents
 
Contributors ................................................... xi
Preface ...................................................... xvii
Volumes in Series ............................................. xix

Section I. Techniques ........................................... 1
1  Laser-Induced Radiation Microbeam Technology and
   Simultaneous Real-Time Fluorescence Imaging in Live Cells .... 3
   Stanley W. Botchway, Pamela Reynolds, Anthony W. Parker,
   and Peter O'Neill
   1  Introduction .............................................. 4
   2  Experimental Procedure ................................... 10
   3  Conclusion/Forward Look .................................. 25
   Acknowledgments ............................................. 26
   References .................................................. 26
2  A Cell Biologist's Guide to High Resolution Imaging ......... 29
   Graeme Ball, Richard M. Parton, Russell S. Hamilton, and
   Ian Davis
   1  Introduction ............................................. 30
   2  Physical Limitations on the Resolution of Conventional
      Microscopy ............................................... 31
   3  Preparations for High Resolution Fluorescence Imaging .... 35
   4  High Resolution Image Data Acquisition and Processing .... 39
   5  Processing ............................................... 43
   6  Analysis of High Resolution Image Data ................... 47
   7  Conclusions .............................................. 51
   Acknowledgments ............................................. 51
   References .................................................. 51
3  Applications of Fluorescence Lifetime Spectroscopy and
   Imaging to Lipid Domains In Vivo ............................ 57
   André E.P. Bastos, Silvia Scolari, Martin Stöckl, and
   Rodrigo F.M. de Almeida
   1  The Challenge of Studying Lipid Domains In Vivo Defied
      by Fluorescence Lifetimes ................................ 58
   2  Importance of Studies In Vitro to Design and
      Rationalize Studies In Vivo .............................. 61
   3  Labeling Cell Membranes In Vivo: Lipophilic Probes
      Versus GFP-Tagged Membrane Proteins ...................... 67
   4  Cuvette Lifetimes and FLIM: In Vivo Applications ......... 70
   Acknowledgments ............................................. 78
   References .................................................. 79
4  Detecting and Tracking Nonfluorescent Nanoparticle Probes
   in Live Cells .............................................. 83
   Gufeng Wang and Ning Fang
   1  Introduction ............................................. 84
   2  Techniques and Tools ..................................... 84
   3  Biological Applications .................................. 90
   4  Cytotoxicity of Nanoparticle Probes ..................... 100
   5  Conclusions and Future Perspective ...................... 101
   Acknowledgments ............................................ 102
   References ................................................. 102
5  Fluorescence Lifetime Microscopy of Tumor Cell Invasion,
   Drug Delivery, and Cytotoxicity ............................ 109
   Gert-jan Bakker, Volker Andresen, Robert M. Hoffman, and
   Peter Friedl
   1  Introduction ............................................ 110
   2  Preparation of Mouse Mammary Tumor Cell Spheroids in a
      Collagen Matrix ......................................... 112
   3  Simultaneous Acquisition of Fluorescence Lifetime and
      Intensity to Monitor DOX Uptake ......................... 113
   4  Monitoring Drug Uptake Kinetics by FLIM ................. 119
   5  Conclusions and Outlook ................................. 123
   Acknowledgment ............................................. 123
   References ................................................. 124
6  Measuring Membrane Protein Dynamics in Neurons Using
   Fluorescence Recovery after Photobleach .................... 127
   Inmaculada M. González-González, Frédéric Jaskуlski,
   Yves Goldberg, Michael C. Ashby, and Jeremy M. Henley
   1  Introduction ............................................ 128
   2  Experimental Parameters and Preparation ................. 130
   3  Equipment ............................................... 133
   4  Establishing FRAP Conditions ............................ 134
   5  A Basic FRAP Protocol ................................... 136
   6  Analysis of FRAP ........................................ 137
7  Acknowledgments References
   Fluorescent Speckle Microscopy in Cultured Cells ........... 147
   Marin Barisic, António J. Pereira, and Helder Maiato
   1  Introduction ............................................ 148
   2  Technical Challenges of FSM ............................. 151
   3  Detailed Methodology for FSM in Cultured Cells .......... 153
   4  Future Challenges Acknowledgments ....................... 159
   References ................................................. 159
8  Green-to-Red Photoconvertible mEosFP-Aided Live Imaging in
   Plants ..................................................... 164
   Jaideep Mathur, Sarah Griffiths, Kiah Barton, and
   Martin H. Schattat
   1  Introduction ............................................ 164
   2  Expression of mEosFP Fusion Proteins in Plants .......... 167
   3  Visualization of mEosFP Probes in Plants ................ 170
   4  Uses of mEosFP Probes in Plants ......................... 173
   5  Post Acquisition Image Processing and Data Creation ..... 177
   Acknowledgments ............................................ 178
   References ................................................. 179
9  Methods for Cell and Particle Tracking ..................... 183
   Erik Meijering, Oleh Dzyubachyk, and lhor Smal
   1  Introduction ............................................ 184
   2  Tracking Approaches ..................................... 185
   3  Tracking Tools .......................................... 187
   4  Tracking Measures ....................................... 192
   5  Tips and Tricks ......................................... 195
   Acknowledgments ............................................ 197
   References ................................................. 197
10 Correlative Light-Electron Microscopy: A Potent Tool
   for the Imaging of Rare or Unique Cellular and Tissue
   Events and Structures ...................................... 201
   Alexander A. Mironov and Galina V. Beznoussenko
   1  Introduction ............................................ 202
   2  Observation of Living Cells and Fixation ................ 203
   3  Immunolabeling with NANOGOLD ............................ 207
   4  Enhancement of Sample Contrast, Sample Locating, and
      Embedding
   5  Identification of the Cell of Interest on EPON Blocks ... 211
   6  Sample Orientation and EM Sectioning from the Very
      First Section ........................................... 212
   7  Picking up Serial Sections with the Empty Slot Grid ..... 214
   EM Analysis ................................................ 216
   Acknowledgments ............................................ 217
   References
11 Optical Techniques for Imaging Membrane Domains in Live
   Cells (Live-Cell Palm of Protein Clustering) ............... 221
   Dylan M. Owen, David Williamson, Astrid Magenau, and
   Katharina Gaus
   1  Introduction ............................................ 222
   2  Sample Preparation and Data Acquisition ................. 224
   3  Data Analysis ........................................... 228
   Acknowledgments ............................................ 234
   References ................................................. 234
12 Single Live Cell Topography and Activity Imaging with the
   Shear-Force-Based Constant-Distance Scanning
   Electrochemical Microscope ................................. 237
   Albert Schulte, Michaela Nebel, and Wolfgang Schuhmann
   1  Introduction ............................................ 238
   2  Shear-Force-Based Constant-Distance Scanning
      Electrochemical Microscopy for Live Cell Studies:
      Apparatus, Probes, and Operation ........................ 240
   3  Selected Applications of SF-CD-SECM Live Cell Studies ... 248
   Acknowledgments ............................................ 253
   References ................................................. 254
13 Visualization of TGN-Endosome Trafficking in Mammalian
   and Drosophila Cells ....................................... 255
   Satoshi Kametaka and Satoshi Waguri
   1  Introduction ............................................ 256
   2  Molecular Tools ......................................... 258
   3  Live-Cell Imaging in Mammalian Cells .................... 260
   4  Live-Cell Imaging in Drosophila Cells ................... 266
   5  Conclusion Remarks ...................................... 269
   References ................................................. 269
14 Live Cell Imaging with Chemical Specificity Using Dual
   Frequency CARS Microscopy .................................. 273
   Lestyn Pope, Wolfgang Langbein, Paola Borri, and Peter
   Watson
   1  Introduction ............................................ 274
   2  "Noninvasive" Live Cell Imaging ......................... 275
   3  Experimental Setup ...................................... 279
   4  Maximizing Collection Efficiency for Live Cell Imaging .. 287
   Acknowledgments ............................................ 289
   References ................................................. 290
15 Imaging Intracellular Protein Dynamics by Spinning Disk
   Confocal Microscopy ........................................ 293
   Samantha Stehbens, Hayley Pemble, Lyndsay Murrow, and
   Torsten Wittmann
   1  Introduction ............................................ 294
   2  Instrument Design ....................................... 298
   3  Combination with Other Imaging Techniques ............... 304
   4  Specimen Preparation .................................... 307
   Acknowledgments ............................................ 312
   References ................................................. 312

Section II. Tools ............................................. 315
16 Visualizing Dynamic Activities of Signaling Enzymes Using
   Genetically Encodable FRET-Based Biosensors: From Designs
   to Applications ............................................ 317
   Xin Zhou, Katie J. Herbst-Robinson, and Jin Zhang
   1  Introduction ............................................ 318
   2  Generalizable Modular Designs ........................... 319
   3  FRET-Based Biosensors for Monitoring Signaling Enzymes .. 324
   4  Example: A-kinase Activity Reporter (AKAR) .............. 332
   5  Summary and Perspectives ................................ 336
   Acknowledgments ............................................ 337
   References ................................................. 337
17 Live-Cell Imaging of Aquaporin-4 Supramolecular Assembly
   and Diffusion .............................................. 341
   A.S. Verkman, Andrea Rossi, and Jonathan M. Crane
   1  Aquaporin-4 (AQP4) and Orthogonal Arrays of Particles ... 342
   2  Approaches to Image AQP4 and OAPs ....................... 343
   3  AQP4 Diffusion and OAPs Studied by Quantum Dot Single
      Particle Tracking ....................................... 345
   4  OAP Dynamics and Structure Studied with GFP-AQP4
      Chimeras ................................................ 347
   5  Single-Molecule Analysis Shows AQP4 Heterotetramers ..... 349
   6  Photobleaching Reveals Post-Golgi Assembly of OAPs ...... 351
   7  Super-Resolution Imaging of AQP4 OAPs ................... 351
   References ................................................. 352
18 Coiled-Coil Tag-Probe Labeling Methods for Live-Cell
   Imaging of Membrane Receptors .............................. 355
   Yoshiaki Yano, Kenichi Kawano, Kaoru Omae, and Katsumi
   Matsuzaki
   1  Introduction ............................................ 356
   2  Various Principles Used for Tag-Probe Labeling .......... 357
   3  Coiled-Coil Tag-Probe Labeling .......................... 360
   4  Applications ............................................ 366
   Acknowledgments ............................................ 368
   References ................................................. 368
19 Monitoring Protein Interactions in Living Cells with
   Fluorescence Lifetime Imaging Microscopy ................... 371
   Yuansheng Sun, Nicole M. Hays, Ammasi Periasamy,
   Michael W. Davidson, and Richard N. Day
   1  Introduction ............................................ 372
   2  FD FLIM Measurements .................................... 376
   3  Measuring Protein-Protein Interactions in Living Cells
      Using FLIM-FRET ......................................... 385
   4  The Strengths and Limitations of FLIM ................... 388
   Acknowledgment ............................................. 389
   References ................................................. 389
20 Open Source Tools for Fluorescent Imaging .................. 393
   Nicholas A. Hamilton
   1  Why Open Source Software? ............................... 394
   2  Open Source Software for Microscopy Imaging and
      Analysis ................................................ 406
   3  The Future .............................................. 414
   Acknowledgments ............................................ 415
   References ................................................. 415
21 Nanoparticle PEBBLE Sensors in Live Cells .................. 419
   Yong-Eun Koo Lee and Raoul Kopelman
   1  Introduction ............................................ 420
   2  PEBBLE Sensor Designs ................................... 424
   3  Preparation/Characterization ............................ 430
   4  Examples ................................................ 431
   5  Summary and Critical Issues ............................. 461
   Acknowledgments ............................................ 462
   References ................................................. 462

Author Index .................................................. 471
Subject Index ................................................. 495


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