ISBN: 3540427686
TITLE: Fluorescence Spectroscopy, Imaging and Probes
AUTHOR: Kraayenhof, Visser, Gerritsen (Eds.)
TOC:

Part 1: Fluorescence Spectroscopy: New Approaches and Probes
1 Advanced Luminescent Labels, Probes and Beads and their Application to Luminescence Bioassay and Imaging 3
O. S. WOLFBEIS, M. BHMER, A. DRKOP, J. ENDERLEIN, M. GRUBER, I. KLIMANT, C. KRAUSE, J. KRNER, G. LIEBSCH, Z. LIN, B. OSWALD, AND M. WU
1.1 Introduction 4
1.2 A General Logic for Designing Fluorescent Cyanine Type Probes and Labels of Defined Color 6
1.3 Diode Laser-excitable Probes for General Protein Detection 8
1.4 Diode Laser-compatible Amino-Reactive Covalent Labels 10
1.5 Diode Laser-assisted Fluorescent Single Molecule Detection of Dyes and Labeled Proteins 11
1.6 New Labels for Flow Cytometric Determination of HSA 14
1.7 Diode Laser-excitable DNA Labels 15
1.8 New Resonance Energy Transfer Gene Assays 16
1.9 Reactive Ruthenium Ligand Complexes as Markers for Bioassays 17
1.10 Diode Laser-excitable Fluorescent Polymer Beads 18
1.11 Polyaniline-Coated Nano-beads (~200 nm in Diameter) with pH-dependent Fluorescence 20
1.12 Phosphorescent Poly(acrylonitrile) Nanospheres (10-100 nm in ) as Markers for Optical Assays 22
1.13 Competitive Binding of Streptavidin to Biotinylated Nanobeads as Studied by Resonance Energy Transfer 25
1.14 Nanobeads as Reference Dyes in Luminescent Lifetime Imaging Using DLR 27
1.15 Phosphorescent Nanospheres for Use in Advanced Time-resolved Multiplexed Bioassays (lambda, tau) 30
1.16 Beads Dyed with a Europium-based Label and Excitable with the 405-nm LED Diode Laser 34
1.17 A Europium(III)-based Probe for Use in Oxidase-Associated Reactions 35
1.17.1 Significance of Probes for Hydrogen Peroxide (HP) 35
1.17.2 A New Probe for Hydrogen Peroxide 35
1.17.3 Glucose Assay Using the Europium Probe 36
1.17.4 Peroxidase Assay Using the Europium Probe 37
1.17.5 Catalase 38
1.17.6 A New Enzyme-linked Immunosorbent Assay (ELISA) 38
References 40
2 Fluorescence Spectral Engineering - Biophysical and Biomedical Applications 43
J. R. LAKOWICZ, I. GRYCZYNSKI, Y. SHEN, J. MALICKA, S. D'AURIA, AND Z. GRYCZYNSKI
2.1 Introduction 44
2.1.1 Fluorescence Spectral Engineering 45
2.1.2 Overview of Metallic Surface Effects on Fluorescence 47
2.1.3 Theory for Fluorophore Metal Interactions 50
2.1.4 Spatial Distribution of Emission Near Metal Surfaces 54
2.1.5 Resonance Energy Transfer 55
2.2 Experimental Results on Fluorophore-metal Interactions 56
2.2.1 Silver Island Films and Experimental Geometry 56
2.2.2 Effects of Silver Island Films on Emission Spectra of Rhodamine B and Rose Bengal 56
2.2.3 Effect of Silver Island Films on Photostability 58
2.2.4 Effects of Silver Island Films on the Lifetime of Rhodamine B and Rose Bengal 59
2.2.5 Effect of Quantum Yield on Silver Island Enhancements 60
2.2.6 Effects of Silver Islands on Intrinsic Protein Fluorescence 61
2.2.7 Effects of Silver Islands on Nucleic Acid Bases and DNA 62
2.2.8 Effects of Silver Islands on Resonance Energy Transfer 63
2.3 Conclusion 65
References 65
3 Fluorescence Nanometrology in Sol-Gels 69
D.J.S. BIRCH, C.D. GEDDES, J. KAROLIN, R. LEISHMAN, AND O.J. ROLINSKI
3.1 Introduction 70
3.2 Sol-gel Chemistry 72
3.3 Anisotropy Theory 75
3.4 FRET Donor-acceptor Distribution Theory 76
3.5 Acidic Hydrogels 78
3.6 Alkaline Hydrogels 79
3.7 Alkoxide Alcogels 80
3.8 Wet Pore Metrology 81
3.9 Conclusions 83
References 84
4 Integrated Supramolecular Systems: From Sensors to Switches 87
J.-P. MALVAL, I. GOSSE, J.-P. MORAND, R. LAPOUYADE
4.1 Introduction 88
4.2 Optical Detection of Ion-Ground State Probe Interaction 90
4.3 Cation Sensing from Fluorescent Photoinduced Intramolecular
Charge Transfer (PICT) Sensors 91
4.4 Optical and Electrochemical Release of Cations 96
4.5 Conclusions 97
References 98
5 Ratiometric Probes: Design and Applications 101
A. P. DEMCHENKO, A. S. KLYMCHENKO, V. G. PIVOVARENKO, AND S. ERCELEN
5.1 Introduction 102
5.2 3-Hydroxyflavones and Other 3-Hydroxychromone Derivatives. New Compounds and Their Properties 103
5.3 A New Level of Sensitivity to Solvent Polarity 104
5.4 Amplification by ESIPT of Electrochromic Effects 106
5.5 Molecular Order and Dynamics in Phospholipid Membranes 107
5.6 Amplification by ESIPT of Site-selective Red Edge Effect 108
5.7 Conclusions 109
References 110
6 Binding of Ethidium to Yeast tRNA^Phe : A New Perspective on an Old Bromide 111
M. TRAMIER, O. HOLUB, J. C. CRONEY, T. ISHI, S. E. SEIFRIED, AND D. M. JAMESON
6.1 Overview 112
6.2 Experimental 112
6.2.1 Sample Preparation 112
6.2.2 Multifrequency Phase and Modulation Fluorometry 112
6.2.3 Three Component Analysis 113
6.3 New Analysis 116
6.4 The Model 118
6.5 Effect of Ionic Strength 118
6.6 Conclusions 119
References 120
7 Experimental Aspects of DNA Computing by Blocking: Use of Fluorescence Techniques for Detection 123
K. A. SCHMIDT, C. V. HENKEL, G. ROZENBERG, AND H. P. SPAINK
7.1 Introduction 124
7.2 Experimental 124
7.3 Results and Discussion 125
7.4 Conclusion 127
References 128
Part 2 Fluorescence Spectroscopy of Single Molecules and Molecular Assemblies
8 Multiparametric Detection of Fluorescence Emitted from Individual Multichromophoric Systems 131
M. COTLET, J. HOFKENS, M. MAUS, AND F. C. DE SCHRYVER
8.1 Introduction 132
8.2 Materials and Methods 133
8.2.1 Sample Preparation 133
8.2.2 Experimental Set-up 133
8.3 Data Processing and Analysis 136
8.3.1 Pulsed Excitation 136
8.3.2 CW Excitation 139
8.4 Results and Discussion 141
8.4.1 Pulsed Excitation 142
8.4.2 CW Excitation 145
8.5 Conclusions 149
References 150
9 Fluorescence Intensity Distribution Analysis (FIDA) and related fluorescence fluctuation techniques: theory and practice 153
P. KASK, C. EGGELING, K. PALO, . METS, M. COLE, AND K. GALL
9.1 Introduction 154
9.2 Instrumentation for Fluorescence Fluctuation Spectroscopy 156
9.3 Assumptions and Conventions 158
9.4 Generating Functions for FIDA and Related Methods 160
9.5 FIDA 162
9.6 2D-FIDA 165
9.7 FIMDA 168
9.8 FILDA 172
9.9 Conclusions 178
References 178
10 Single Molecule Reactions of the Enzyme LDH and of Restriction Endonucleases in the Fluorescence Microscope 183
B. NASANSHARGAL, B. SCHFER, AND K. O. GREULICH
10.1 Introduction 184
10.2 Femtodroplets and the Poisson Statistics 185
10.3 From Concentrations to Intermolecular Distances 186
10.4 Why Fluorescence Microscopy? 187
10.5 Single Molecule Enzyme Reactions with Small Substrates 187
10.6 Restriction Endonuclease Reactions 191
10.7 Conclusions 194
References 195
11 Monitoring gamma-Subunit Movement in Reconstituted Single EF_0 F_1 ATP Synthase by Fluorescence Resonance Energy Transfer 197
M. BRSCH, M. DIEZ, B. ZIMMERMANN, R. REUTER, AND P. GRBER
11.1 Introduction 198
11.2 Visualizing Intersubunit Rotation 198
11.3 FRET-labeled F_0 F_1 ATP Synthase EF_0-b64-TMR-F_1-gamma106-Cy5 200
11.3.1 Synthezising ATP with Reconstituted EF:_0-b64-TMR- F_1-gamma106-Cy5 201
11.3.2 Set-up for Single Enzyme FRET Analysis 201
11.3.3 Discrimination of Three gamma-Subunit Positions with AMPPNP 203
11.3.4 Unidirectional Motion of the gamma-Subunit with ATP 205
11.4 Conclusions 206
References 207
Part 3 Application of Fluorescence in Biological Membrane and Enzyme Studies
12 Application of the Wavelength-selective Fluorescence Approach to Monitor Membrane Organization and Dynamics 211
A. CHATTOPADHYAY
12.1 Introduction 212
12.2 Red Edge Excitation Shift (REES) 212
12.3 The Wavelength-selective Fluorescence Approach 214
12.4 The Wavelength-selective Fluorescence Approach: A Novel Tool to Monitor Organization and Dynamics of the Membrane Interfacial Region 214
12.5 Wavelength-selective Fluorescence as a Membrane Dipstick 217
12.6 Application of the Wavelength-selective Fluorescence Approach to Membrane Peptides and Proteins 218
12.7 Wavelength-selective Fluorescence in Micelles 220
12.8 Conclusions 221
References 221
13 Fluorescence Approaches for the Characterization of the Peripheral Membrane Binding of Proteins Applied for the Blood Coagulation Protein Prothrombin 225
R. HUTTERER AND M. HOF
13.1 Introduction 226
13.2 Prothrombin Binding to Negatively Charged Membrane Surface Characterized by Protein Fluorescence 227
13.2.1 Intrinsic Protein Fluorescence: Picosecond Tryptophan Fluorescence of Membrane-bound Prothrombin Fragment 1 (F1) 227
13.2.2 Overview on Investigations Applying Fluorescently Labeled Prothrombin 231
13.3 Prothrombin-induced Changes in the Organization of Phospholipid Bilayers 233
13.3.1 Solvent Relaxation (SR) 233
13.3.1.1 Solvent Relaxation Probed by the Headgroup Labels Prodan and Patman 233
13.3.1.2 Influence of Prothrombin and its Fragment 1 on the Phospolipid Headgroup Organization 235
13.3.2 Fluorescence Anisotropy: Influence of Prothrombin and its Fragment 1 on PC/PS Membrane Order 237
13.3.3 Pyrene Fluorescence: Influence of Fragment 1 on Membrane Order Monitored by the Excimer/Monomer Ratio 237
13.4 Conclusions 238
References 239
14 Assessment of Membrane Fluidity in Individual Yeast Cells by Laurdan Generalised Polarisation and Multi-photon Scanning Fluorescence Microscopy 241
R. P. LEARMONTH AND E. GRATTON
14.1 Introduction 242
14.1.1 Yeast Membrane Fluidity 242
14.1.2 Multi-photon Scanning Fluorescence Microscopy 243
14.1.3 Determination of Membrane Fluidity Using Laurdan Generalized Polarization 244
14.2 Materials and Methods 244
14.3 Results and Discussion 246
14.4 Conclusions 250
References 251
15 Formation of Higher Order Signal Transduction Complexes as Seen by Fluorescence Spectroscopy 253
L. DOWAL AND S. SCARLATA
15.1 Introduction 254
15.2 Experimental 256
15.2.1 Membrane Binding 257
15.2.2 Interactions of PLC beta and G Protein Subunits on Membrane Surface 257
15.2.3 PLC beta_2-G beta gammaAssociations on Membrane Surfaces 258
15.2.4 Effects of Lipid Rafts on Protein Association 259
References 260
16 Mechanisms of the Modulation of Membrane Interfacial Enzyme Catalysis by Non-lamellar Forming Lipids: Comparison with the Behavior of a Fluorescent Probe in Membranes 263
R. M. EPAND, R. CORNELL, S. M.A. DAVIES, AND R. KRAAYENHOF
16.1 Introduction 264
16.1.1 Specific vs. Non-specific Modulation of Protein Activity 264
16.1.2 Non-lamellar Forming Lipids 265
16.2 Specific Protein Examples 266
16.2.1 Protein Kinase C 266
16.2.2 Phosphocholine Cytidylyltransferase 267
16.3 Molecular Mechanisms of Enzyme Activation by Non-lamellar Forming Lipids 268
16.3.1 Curvature Strain 268
16.3.2 Lateral Pressure Profile 269
16.3.3 Other Mechanisms 269
16.4 Criteria for the Role of Membrane Curvature Strain 269
16.4.1 Cubic Phases 269
16.4.2 Homologous Lipids 271
16.5 Relationship of the Properties of a Fluorescent Interfacial Membrane Probe 271
16.6 Comparison of the Mechanisms of Activation of PKC and CT by Non-lamellar Forming Lipids 272
16.7 Conclusions 273
References 274
17 Emission Spectroscopy of Complex Formation between Escherichia coli Purine Nucleoside Phosphorylase (PNP) and Identified Tautomeric Species of Formycin Inhibitors Resolves Ambiguities Found in Crystallographic Studies 277
B. KIERDASZUK
17.1 Introduction 278
17.2 Formycin A and its N-methyl Analogs, Specific Inhibitors of E. coli PNP 280
17.3 Ambiguities Found in the Crystallographic Structure of Enzyme-Ligand Complex 281
17.4 Solution Structure of Inhibitors Bound by the Enzyme 283
17.4.1 Tautomeric Equilibria, and Absorption and Emission Spectra of the Tautomeric Species in Solution 283
17.4.2 Shifts Between Absorption and Emission Spectra of the Enzyme and Ligands 286
17.4.3 Effect of Binding of the Identified Tautomeric Species on their Fluorescence and Phosphorescence 287
17.5 Conclusions 291
References 292
Part 4 Microscopic Imaging Techniques and their Application for the Study of Living Cells
18 Fluorescence Lifetime Imaging Implemented with Resonant Galvanometer Scanners 297
J. J. BIRMINGHAM
18.1 Introduction 298
18.2 Theory 299
18.3 Simulations 305
18.4 Experimental Set-up 308
18.5 Results 311
18.6 Conclusions 314
References 315
19 Spectral Imaging of Single CdSe/ZnS Quantum Dots Employing Spectrally- and Time-resolved Confocal Microscopy 317
W.G.J.H.M. VAN SARK, P.L.T.M. FREDERIX, M.A.H. ASSELBERGS, D.J. VAN DEN HEUVEL, A. MEIJERINK, AND H. C. GERRITSEN
19.1 Introduction 318
19.2 Experimental 319
19.2.1 Spectrograph-CLSM Set-up 319
19.2.1.1 Description 319
19.2.1.2 Performance 322
19.2.2 QD Synthesis and Characterization 324
19.3 Results and Discussion 325
19.4 Conclusion 332
References 332
20 Imaging of Oxidative Stress in Plant Cells by Quantitative Fluorescence Microscopy and Spectroscopy 337
J. W. BORST, M. A. USKOVA, N. V. VISSER, AND A. J. W. G. VISSER
20.1 Introduction 338
20.2 Experimental Procedures 340
20.2.1 Tobacco BY-2 Cells 340
20.2.2 Preparation of Cell Suspension 340
20.2.3 Protoplast Isolation 341
20.2.4 Labeling Cells with BP-C11 341
20.2.5 Confocal Microscopy 341
20.2.6 Steady-state Fluorescence 342
20.3 Results and Discussion 342
20.3.1 Measurements and Imaging of Oxidative Stress in Tobacco BY-2 Cells 342
20.3.2 Effect of Inhibitors and ROS Scavengers on the Hydrogen Peroxide Production in BY-2 Cells 344
20.4 Conclusions 346
References 347
21 The Biomedical Use of Rescaling Procedures in Optical Biopsy and Optical Molecular Imaging 349
O. MINET, J. BEUTHAN, K. LICHA, AND C. MAHNKE
21.1 Introduction 350
21.2 Method of Rescaling 350
21.3 Biomedical Examples 352
21.3.1 Optical Biopsy in the UV Range Using Endogenous Chromophores 352
21.3.1.1 Experiment 353
21.3.1.2 Results 354
21.3.2 Optical Molecular Imaging in the NIR Range Using Exogenous Contrast Agents 354
21.3.2.1 Experiment 356
21.3.2.2 Results 357
21.4 Discussion and Conclusions 358
References 359
22 Looking into a Living Cell 361
M. VAN BORREN, N. R. BRADY, J. RAVELSLOOT, AND H. V. WESTERHOFF
22.1 Introduction 362
22.2 How to Choose your Fluorescent Indicator 362
22.2.1 Process of Interest 362
22.2.2 Ratiometric Dyes 363
22.2.3 Buffering Power 364
22.2.4 Photo-toxicity 364
22.3 Considerations Concerning the Experimental Approach 364
22.3.1 Selective Loading 364
22.3.2 Additional Loading Techniques 366
22.3.3 The Imaging System 366
22.4 Examples of Fluorescence Microscopy in Living Cells 366
22.4.1 Detection of the Ca^2+ Ion 366
22.4.2 Determination of Intracellular pH 367
22.4.3 Mitochondrial Energetics 368
22.4.4 Reactive Oxygen Species (ROS) 370
22.4.5 Autofluorescence: Marker of Redox State 371
22.5 Conclusions 371
References 371
23 Expression of Multicolor Fluorescent Fusion Proteins in Zebrafish Cell Cultures: A Versatile Tool in Cell Biology 373
C. K. D. BREEK, F. VAN IREN, S. E. WIJTING, N. STUURMAN, AND H. P. SPAINK
23.1 Introduction 374
23.2 Zebrafish Cell Lines 374
23.3 Microscopical Analysis 376
23.4 Analysis of Transfected Cell Lines 377
23.5 Conclusions 379
References 379
Subject Index 381
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