Handbook of fluorescence spectroscopy and imaging: from single molecules to ensembles
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| Main Authors: | , , |
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| Format: | Book |
| Language: | English |
| Published: |
Weinheim
Wiley-VCH
2011
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| Subjects: | |
| Online Access: | Inhaltstext Inhaltsverzeichnis |
| Physical Description: | IX, 281 S. Ill., graph. Darst. |
| ISBN: | 9783527316694 3527316698 |
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| adam_text |
Titel: Handbook of fluorescence spectroscopy and imaging
Autor: Sauer, Markus
Jahr: 2011
Contents
Preface IX
1 Basic Principles of Fluorescence Spectroscopy 1
1.1 Absorption and Emission of Light 1
1.2 Spectroscopic Transition Strengths 5
1.3 Lambert-Beer Law and Absorption Spectroscopy 7
1.4 Fluorophore Dimerization and Isosbestic Points 9
1.5 Franck-Condon Principle 12
1.6 Temperature Effects on Absorption and Emission Spectra 15
1.7 Fluorescence and Competing Processes 17
1.8 Stokes Shift, Solvent Relaxation, and Solvatochroism 20
1.9 Fluorescence Quantum Yield and Lifetime 22
1.10 Fluorescence Anisotropy 27
References 29
2 Fluorophores and Fluorescent Labels 31
2.1 Natural Fluorophores 31
2.2 Organic Fluorophores 35
2.3 Different Fluorophore Classes 38
2.4 Multichromophoric Labels 49
2.5 Nanocrystals 52
References 56
3 Fluorophore Labeling for Single-Molecule Fluorescence
Spectroscopy (SMFS) 61
3.1 In Vitro Fluorescence Labeling 61
3.2 Fluorescence Labeling in Living Cells 69
References 80
4 Fluorophore Selection for Single-Molecule Fluorescence Spectroscopy
(SMFS) and Photobleaching Pathways 85
References 91
5 Fluorescence Correlation Spectroscopy 93
5.1 Introduction 93
5.2 Optical Set-Up 98
5.3 Data Acquisition and Evaluation 99
5.4 Milliseconds to Seconds: Diffusion and Concentration 103
5.4.1 Single-Focus FCS 104
5.4.2 Dual-Focus FCS 111
5.5 Nanoseconds to Microseconds: Photophysics, Conformational
Fluctuations, Binding Dynamics 120
5.6 Picoseconds to Nanoseconds: Rotational Diffusion
and Fluorescence Antibunching 122
5.6.1 Antibunching 122
5.6.2 Rotational Diffusion 125
5.7 Fluorescence Lifetime Correlation Spectroscopy 135
5.8 Conclusion 143
References 143
6 Excited State Energy Transfer 147
6.1 Introduction 147
6.2 Theory of (Förster) Energy Transfer 148
6.2.1 Mechanism and Mathematical Formalism of FRET 148
6.2.2 Measuring FRET Efficiencies Through Excited-State Lifetimes 153
6.2.3 Spin Rules for FRET 154
6.2.4 Homo-FRET and FRET-Induced Depolarization 154
6.3 Experimental Approach for Single-Pair F RET-Experiments 157
6.3.1 Single-Laser Excitation 157
6.3.2 Alternating-Laser Excitation (ALEX) 160
6.4 Examples and Applications of FRET 161
6.4.1 FRET Processes in Bulk Experiments 162
6.4.1.1 FRET-Based Molecular Biosensors 162
6.4.1.2 Energy Hopping and Trapping in Chromophore-Substituted
Polyphenylene Dendrimers 164
6.4.2 Single-Molecule Observation of FRET 168
6.4.2.1 Light-Harvesting Systems: Phycobilisomes and Allophycocyanins 168
6.4.2.2 Hairpin Ribozyme Dynamics and Activity 179
6.4.2.3 Protein (Un)folding and Dynamics 180
References 183
7 Photoinduced Electron Transfer (PET) Reactions 189
7.1 Fluorescence Quenching by PET 189
7.2 Single-Molecule Fluorescence Spectroscopy to Study PET 192
7.3 Single-Molecule Sensitive Fluorescence Sensors Based on PET 199
7.4 PET Reporter System 202
7.5 Monitoring Conformational Dynamics and Protein Folding
by PET 205
7.6 Biological and Diagnostic Applications 209
References 215
8 Super-Resolution Fluorescence Imaging 219
8.1 Diffraction Barrier of Optical Microscopy 219
8.2 Multi-Photon and Structured Illumination Microscopy 221
8.3 Stimulated Emission Depletion 223
8.4 Single-Molecule Based Photoswitching Microscopy 226
8.5 Background and Principles of Single-Molecule Based
Photoswitching Microscopy Methods 229
8.6 Temporal Resolution of Super-Resolution Imaging Methods 236
References 237
9 Single-Molecule Enzymatics 241
9.1 Introduction: Why Study Enzymes on a Single-Molecule Level? 241
9.2 Biochemical Principles of Enzymatic Activity: the Michaelis-Menten
Model 242
9.3 "Looking" at Individual Enzymes 243
9.3.1 Single-Enzyme Studies and Kinetics 244
9.3.1.1 Space-Resolved, but Time-Averaged Single Enzyme Assays 244
9.3.1.2 Single-Turnover Experiments: Space- and Time-Resolved
Enzyme Assays 248
9.3.1.3 Results: Revision of the Classical Michaelis-Menten Model 255
9.3.2 Conformational Dynamics 259
9.3.3 Single-Molecule DNA Sequencing 260
9.3.4 Shedding Light on Single-Enzyme Mechanisms 262
9.3.4.1 Movement of Molecular Motor Enzymes on Actin Filaments 263
9.3.4.2 Lipase-Catalyzed Hydrolysis of Phospholipid Bilayers 264
9.4 Data Analysis of Fluorescence Intensity Time Traces
of Single-Turnover Experiments 264
9.4.1 Threshold Method 264
9.4.2 Autocorrelation Analysis 267
9.5 Conclusions 267
References 268
Index 273 |
| adam_txt |
Titel: Handbook of fluorescence spectroscopy and imaging
Autor: Sauer, Markus
Jahr: 2011
Contents
Preface IX
1 Basic Principles of Fluorescence Spectroscopy 1
1.1 Absorption and Emission of Light 1
1.2 Spectroscopic Transition Strengths 5
1.3 Lambert-Beer Law and Absorption Spectroscopy 7
1.4 Fluorophore Dimerization and Isosbestic Points 9
1.5 Franck-Condon Principle 12
1.6 Temperature Effects on Absorption and Emission Spectra 15
1.7 Fluorescence and Competing Processes 17
1.8 Stokes Shift, Solvent Relaxation, and Solvatochroism 20
1.9 Fluorescence Quantum Yield and Lifetime 22
1.10 Fluorescence Anisotropy 27
References 29
2 Fluorophores and Fluorescent Labels 31
2.1 Natural Fluorophores 31
2.2 Organic Fluorophores 35
2.3 Different Fluorophore Classes 38
2.4 Multichromophoric Labels 49
2.5 Nanocrystals 52
References 56
3 Fluorophore Labeling for Single-Molecule Fluorescence
Spectroscopy (SMFS) 61
3.1 In Vitro Fluorescence Labeling 61
3.2 Fluorescence Labeling in Living Cells 69
References 80
4 Fluorophore Selection for Single-Molecule Fluorescence Spectroscopy
(SMFS) and Photobleaching Pathways 85
References 91
5 Fluorescence Correlation Spectroscopy 93
5.1 Introduction 93
5.2 Optical Set-Up 98
5.3 Data Acquisition and Evaluation 99
5.4 Milliseconds to Seconds: Diffusion and Concentration 103
5.4.1 Single-Focus FCS 104
5.4.2 Dual-Focus FCS 111
5.5 Nanoseconds to Microseconds: Photophysics, Conformational
Fluctuations, Binding Dynamics 120
5.6 Picoseconds to Nanoseconds: Rotational Diffusion
and Fluorescence Antibunching 122
5.6.1 Antibunching 122
5.6.2 Rotational Diffusion 125
5.7 Fluorescence Lifetime Correlation Spectroscopy 135
5.8 Conclusion 143
References 143
6 Excited State Energy Transfer 147
6.1 Introduction 147
6.2 Theory of (Förster) Energy Transfer 148
6.2.1 Mechanism and Mathematical Formalism of FRET 148
6.2.2 Measuring FRET Efficiencies Through Excited-State Lifetimes 153
6.2.3 Spin Rules for FRET 154
6.2.4 Homo-FRET and FRET-Induced Depolarization 154
6.3 Experimental Approach for Single-Pair F RET-Experiments 157
6.3.1 Single-Laser Excitation 157
6.3.2 Alternating-Laser Excitation (ALEX) 160
6.4 Examples and Applications of FRET 161
6.4.1 FRET Processes in Bulk Experiments 162
6.4.1.1 FRET-Based Molecular Biosensors 162
6.4.1.2 Energy Hopping and Trapping in Chromophore-Substituted
Polyphenylene Dendrimers 164
6.4.2 Single-Molecule Observation of FRET 168
6.4.2.1 Light-Harvesting Systems: Phycobilisomes and Allophycocyanins 168
6.4.2.2 Hairpin Ribozyme Dynamics and Activity 179
6.4.2.3 Protein (Un)folding and Dynamics 180
References 183
7 Photoinduced Electron Transfer (PET) Reactions 189
7.1 Fluorescence Quenching by PET 189
7.2 Single-Molecule Fluorescence Spectroscopy to Study PET 192
7.3 Single-Molecule Sensitive Fluorescence Sensors Based on PET 199
7.4 PET Reporter System 202
7.5 Monitoring Conformational Dynamics and Protein Folding
by PET 205
7.6 Biological and Diagnostic Applications 209
References 215
8 Super-Resolution Fluorescence Imaging 219
8.1 Diffraction Barrier of Optical Microscopy 219
8.2 Multi-Photon and Structured Illumination Microscopy 221
8.3 Stimulated Emission Depletion 223
8.4 Single-Molecule Based Photoswitching Microscopy 226
8.5 Background and Principles of Single-Molecule Based
Photoswitching Microscopy Methods 229
8.6 Temporal Resolution of Super-Resolution Imaging Methods 236
References 237
9 Single-Molecule Enzymatics 241
9.1 Introduction: Why Study Enzymes on a Single-Molecule Level? 241
9.2 Biochemical Principles of Enzymatic Activity: the Michaelis-Menten
Model 242
9.3 "Looking" at Individual Enzymes 243
9.3.1 Single-Enzyme Studies and Kinetics 244
9.3.1.1 Space-Resolved, but Time-Averaged Single Enzyme Assays 244
9.3.1.2 Single-Turnover Experiments: Space- and Time-Resolved
Enzyme Assays 248
9.3.1.3 Results: Revision of the Classical Michaelis-Menten Model 255
9.3.2 Conformational Dynamics 259
9.3.3 Single-Molecule DNA Sequencing 260
9.3.4 Shedding Light on Single-Enzyme Mechanisms 262
9.3.4.1 Movement of Molecular Motor Enzymes on Actin Filaments 263
9.3.4.2 Lipase-Catalyzed Hydrolysis of Phospholipid Bilayers 264
9.4 Data Analysis of Fluorescence Intensity Time Traces
of Single-Turnover Experiments 264
9.4.1 Threshold Method 264
9.4.2 Autocorrelation Analysis 267
9.5 Conclusions 267
References 268
Index 273 |
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| spelling | Sauer, Markus 1965- Verfasser (DE-588)143429698 aut Handbook of fluorescence spectroscopy and imaging from single molecules to ensembles Markus Sauer ; Johan Hofkens ; Jörg Enderlein Weinheim Wiley-VCH 2011 IX, 281 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Fluoreszenzspektroskopie (DE-588)4017701-4 gnd rswk-swf Fluoreszenzspektroskopie (DE-588)4017701-4 s DE-604 Hofkens, Johan Verfasser (DE-588)143429701 aut Enderlein, Jörg Verfasser (DE-588)113253354 aut text/html http://deposit.dnb.de/cgi-bin/dokserv?id=2974365&prov=M&dok_var=1&dok_ext=htm Inhaltstext HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016769720&sequence=000004&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Sauer, Markus 1965- Hofkens, Johan Enderlein, Jörg Handbook of fluorescence spectroscopy and imaging from single molecules to ensembles Fluoreszenzspektroskopie (DE-588)4017701-4 gnd |
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| title | Handbook of fluorescence spectroscopy and imaging from single molecules to ensembles |
| title_auth | Handbook of fluorescence spectroscopy and imaging from single molecules to ensembles |
| title_exact_search | Handbook of fluorescence spectroscopy and imaging from single molecules to ensembles |
| title_exact_search_txtP | Handbook of fluorescence spectroscopy and imaging from single molecules to ensembles |
| title_full | Handbook of fluorescence spectroscopy and imaging from single molecules to ensembles Markus Sauer ; Johan Hofkens ; Jörg Enderlein |
| title_fullStr | Handbook of fluorescence spectroscopy and imaging from single molecules to ensembles Markus Sauer ; Johan Hofkens ; Jörg Enderlein |
| title_full_unstemmed | Handbook of fluorescence spectroscopy and imaging from single molecules to ensembles Markus Sauer ; Johan Hofkens ; Jörg Enderlein |
| title_short | Handbook of fluorescence spectroscopy and imaging |
| title_sort | handbook of fluorescence spectroscopy and imaging from single molecules to ensembles |
| title_sub | from single molecules to ensembles |
| topic | Fluoreszenzspektroskopie (DE-588)4017701-4 gnd |
| topic_facet | Fluoreszenzspektroskopie |
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