Molecular fluorescence : principles and applications / Bernard Valeur.

Format:

Book

Author/Creator:

Valeur, Bernard, 1944-

Contributor:

Alumni and Friends Memorial Book Fund.

Language:

English

Subjects (All):

Fluorescence spectroscopy.

Physical Description:

xiv, 387 pages : illustrations (some color) ; 25 cm

Place of Publication:

Weinheim ; New York : Wiley-VCH, 2002.

Summary:

Today, fluorescence spectroscopy is an important tool of investigation in many areas. In analytical sciences, its advantage is extremely high sensitivity and selectivity - even single molecules can be detected - and it achieves a high spatial resolution and time resolution in combination with microscopic techniques or laser techniques, respectively. In material sciences this is used to study structure and dynamics of surfaces. Particularly in the areas of biochemistry and molecular genetics, fluorescence spectroscopy has become a dominating technique. Together with the latest imaging techniques, fluorescence spectroscopy allows a real-time observation of the dynamics of intact biological systems with an unprecedented resolution. This book offers a comprehensive introduction to and survey of fluorescence spectroscopy. It is written for newcomers and active researchers alike who are learning to apply fluorescence methods in the areas of chemistry, physical chemistry, polymers, materials, colloids, biochemistry, biology, medical and pharmaceutical research.

Contents:

1.1 What is luminescence? 3

1.2 A brief history of fluorescence and phosphorescence 5

1.3 Fluorescence and other de-excitation processes of excited molecules 8

1.4 Fluorescent probes 11

1.5 Molecular fluorescence as an analytical tool 15

1.6 Ultimate spatial and temporal resolution: femtoseconds, femtoliters, femtomoles and single-molecule detection 16

2 Absorption of UV-visible light 20

2.1 Types of electronic transitions in polyatomic molecules 20

2.2 Probability of transitions. The Beer-Lambert Law. Oscillator strength 23

2.3 Selection rules 30

2.4 The Franck-Condon principle 30

3 Characteristics of fluorescence emission 34

3.1 Radiative and non-radiative transitions between electronic states 34

3.1.1 Internal conversion 37

3.1.2 Fluorescence 37

3.1.3 Intersystem crossing and subsequent processes 38

3.1.3.1 Intersystem crossing 41

3.1.3.2 Phosphorescence versus non-radiative de-excitation 41

3.1.3.3 Delayed fluorescence 41

3.1.3.4 Triplet-triplet transitions 42

3.2 Lifetimes and quantum yields 42

3.2.1 Excited-state lifetimes 42

3.2.2 Quantum yields 46

3.2.3 Effect of temperature 48

3.3 Emission and excitation spectra 48

3.3.1 Steady-state fluorescence intensity 48

3.3.2 Emission spectra 50

3.3.3 Excitation spectra 52

3.3.4 Stokes shift 54

3.4 Effects of molecular structure on fluorescence 54

3.4.1 Extent of [pi]-electron system. Nature of the lowest-lying transition 54

3.4.2 Substituted aromatic hydrocarbons 56

3.4.2.1 Internal heavy atom effect 56

3.4.2.2 Electron-donating substituents: -OH, -OR, -NHR, -NH[subscript 2] 56

3.4.2.3 Electron-withdrawing substituents: carbonyl and nitro compounds 57

3.4.2.4 Sulfonates 58

3.4.3 Heterocyclic compounds 59

3.4.4 Compounds undergoing photoinduced intramolecular charge transfer (ICT) and internal rotation 62

3.5 Environmental factors affecting fluorescence 67

3.5.1 Homogeneous and inhomogeneous broadening. Red-edge effects 67

3.5.2 Solid matrices at low temperature 68

3.5.3 Fluorescence in supersonic jets 70

4 Effects of intermolecular photophysical processes on fluorescence emission 72

4.2 Overview of the intermolecular de-excitation processes of excited molecules leading to fluorescence quenching 74

4.2.1 Phenomenological approach 74

4.2.2 Dynamic quenching 77

4.2.2.1 Stern-Volmer kinetics 77

4.2.2.2 Transient effects 79

4.2.3 Static quenching 84

4.2.3.1 Sphere of effective quenching 84

4.2.3.2 Formation of a ground-state non-fluorescent complex 85

4.2.4 Simultaneous dynamic and static quenching 86

4.2.5 Quenching of heterogeneously emitting systems 89

4.3 Photoinduced electron transfer 90

4.4 Formation of excimers and exciplexes 94

4.5 Photoinduced proton transfer 99

4.5.1 General equations 100

4.5.2 Determination of the excited-state pK 103

4.5.2.1 Prediction by means of the Forster cycle 103

4.5.2.2 Steady-state measurements 105

4.5.2.3 Time-resolved experiments 106

4.5.3 pH dependence of absorption and emission spectra 106

4.6 Excitation energy transfer 110

4.6.1 Distinction between radiative and non-radiative transfer 110

4.6.2 Radiative energy transfer 110

4.6.3 Non-radiative energy transfer 113

5 Fluorescence polarization. Emission anisotropy 125

5.1 Characterization of the polarization state of fluorescence (polarization ratio, emission anisotropy) 127

5.1.1 Excitation by polarized light 129

5.1.1.1 Vertically polarized excitation 129

5.1.1.2 Horizontally polarized excitation 130

5.1.2 Excitation by natural light 130

5.2 Instantaneous and steady-state anisotropy 131

5.2.1 Instantaneous anisotropy 131

5.2.2 Steady-state anisotropy 132

5.3 Additivity law of anisotropy 132

5.4 Relation between emission anisotropy and angular distribution of the emission transition moments 134

5.5 Case of motionless molecules with random orientation 135

5.5.1 Parallel absorption and emission transition moments 135

5.5.2 Non-parallel absorption and emission transition moments 138

5.6 Effect of rotational Brownian motion 140

5.6.1 Free rotations 143

5.6.2 Hindered rotations 150

5.7 Applications 151

6 Principles of steady-state and time-resolved fluorometric techniques 155

6.1 Steady-state spectrofluorometry 155

6.1.1 Operating principles of a spectrofluorometer 156

6.1.2 Correction of excitation spectra 158

6.1.3 Correction of emission spectra 159

6.1.4 Measurement of fluorescence quantum yields 159

6.1.5 Problems in steady-state fluorescence measurements: inner filter effects and polarization effects 161

6.1.6 Measurement of steady-state emission anisotropy. Polarization spectra 165

6.2 Time-resolved fluorometry 167

6.2.1 General principles of pulse and phase-modulation fluorometries 167

6.2.2 Design of pulse fluorometers 173

6.2.2.1 Single-photon timing technique 173

6.2.2.2 Stroboscopic technique 176

6.2.2.3 Other techniques 176

6.2.3 Design of phase-modulation fluorometers 177

6.2.3.1 Phase fluorometers using a continuous light source and an electro-optic modulator 178

6.2.3.2 Phase fluorometers using the harmonic content of a pulsed laser 180

6.2.4 Problems with data collection by pulse and phase-modulation fluorometers 180

6.2.4.1 Dependence of the instrument response on wavelength. Color effect 180

6.2.4.2 Polarization effects 181

6.2.4.3 Effect of light scattering 181

6.2.5 Data analysis 181

6.2.5.1 Pulse fluorometry 181

6.2.5.2 Phase-modulation fluorometry 182

6.2.5.3 Judging the quality of the fit 183

6.2.5.4 Global analysis 184

6.2.5.5 Complex fluorescence decays. Lifetime distributions 185

6.2.6 Lifetime standards 186

6.2.7 Time-dependent anisotropy measurements 189

6.2.7.1 Pulse fluorometry 189

6.2.7.2 Phase-modulation fluorometry 192

6.2.8 Time-resolved fluorescence spectra 192

6.2.9 Lifetime-based decomposition of spectra 194

6.2.10 Comparison between pulse and phase fluorometries 195

6.3 Appendix: Elimination of polarization effects in the measurement of fluorescence intensity and lifetime 196

7 Effect of polarity on fluorescence emission. Polarity probes 200

7.1 What is polarity? 200

7.2 Empirical scales of solvent polarity based on solvatochromic shifts 202

7.2.1 Single-parameter approach 202

7.2.2 Multi-parameter approach 204

7.3 Photoinduced charge transfer (PCT) and solvent relaxation 206

7.4 Theory of solvatochromic shifts 208

7.5 Examples of PCT fluorescent probes for polarity 213

7.6 Effects of specific interactions 217

7.6.1 Effects of hydrogen bonding on absorption and fluorescence spectra 218

7.6.2 Examples of the effects of specific interactions 218

7.6.3 Polarity-induced inversion of n-[pi] and [pi]-[pi] states 221

7.7 Polarity-induced changes in vibronic bands. The Py scale of polarity 222

8 Microviscosity, fluidity, molecular mobility. Estimation by means of fluorescent probes 226

8.1 What is viscosity? Significance at a microscopic level 226

8.2 Use of molecular rotors 230

8.3 Methods based on intermolecular quenching or intermolecular excimer formation 232

8.4 Methods based on intramolecular excimer formation 235

8.5 Fluorescence polarization method 237

8.5.1 Choice of probes 237

8.5.2 Homogeneous isotropic media 240

8.5.3 Ordered systems 242

8.5.4 Practical aspects 242

9 Resonance energy transfer and its applications 247

9.2 Determination of distances at a supramolecular level using RET 249

9.2.1 Single distance between donor and acceptor 249

9.2.2 Distributions of distances in donor-acceptor pairs 254

9.3 RET in ensembles of donors and acceptors 256

9.3.1 RET in three dimensions. Effect of viscosity 256

9.3.2 Effects of dimensionality on RET 260

9.3.3 Effects of restricted geometries on RET 261

9.4 RET between like molecules.

Excitation energy migration in assemblies of chromophores 264

9.4.1 RET within a pair of like chromophores 264

9.4.2 RET in assemblies of like chromophores 265

9.4.3 Lack of energy transfer upon excitation at the red-edge of the absorption spectrum (Weber's red-edge effect) 265

9.5 Overview of qualitative and quantitative applications of RET 268

10 Fluorescent molecular sensors of ions and molecules 273

10.2 pH sensing by means of fluorescent indicators 276

10.2.2 The main fluorescent pH indicators 283

10.2.2.1 Coumarins 283

10.2.2.2 Pyranine 283

10.2.2.3 Fluorescein and its derivatives 283

10.2.2.4 SNARF and SNAFL 284

10.2.2.5 PET (photoinduced electron transfer) pH indicators 286

10.3 Fluorescent molecular sensors of cations 287

10.3.1 General aspects 287

10.3.2 PET (photoinduced electron transfer) cation sensors 292

10.3.2.2 Crown-containing PET sensors 293

10.3.2.3 Cryptand-based PET sensors 294

10.3.2.4 Podand-based and chelating PET sensors 294

10.3.2.5 Calixarene-based PET sensors 295

10.3.2.6 PET sensors involving excimer formation 296

10.3.2.7 Examples of PET sensors involving energy transfer 298

10.3.3 Fluorescent PCT (photoinduced charge transfer) cation sensors 298

10.3.3.2 PCT sensors in which the bound cation interacts with an electron-donating group 299

10.3.3.3 PCT sensors in which the bound cation interacts with an electron-withdrawing group 305

10.3.4 Excimer-based cation sensors 308

10.3.5.1 Oxyquinoline-based cation sensors 310

10.3.5.2 Further calixarene-based fluorescent sensors 313

10.4 Fluorescent molecular sensors of anions 315

10.4.1 Anion sensors based on collisional quenching 315

10.4.2 Anion sensors containing an anion receptor 317

10.5 Fluorescent molecular sensors of neutral molecules and surfactants 322

10.5.1 Cyclodextrin-based fluorescent sensors 323

10.5.2 Boronic acid-based fluorescent sensors 329

10.5.3 Porphyrin-based fluorescent sensors 329

10.6 Towards fluorescence-based chemical sensing devices 333

Appendix A. Spectrophotometric and spectrofluorometric pH titrations 337

Appendix B. Determination of the stoichiometry and stability constant of metal complexes from spectrophotometric or spectrofluorometric titrations 339

11 Advanced techniques in fluorescence spectroscopy 351

11.1 Time-resolved fluorescence in the femtosecond time range: fluorescence up-conversion technique 351

11.2 Advanced fluorescence microscopy 353

11.2.1 Improvements in conventional fluorescence microscopy 353

11.2.1.1 Confocal fluorescence microscopy 354

11.2.1.2 Two-photon excitation fluorescence microscopy 355

11.2.1.3 Near-field scanning optical microscopy (NSOM) 356

11.2.2 Fluorescence lifetime imaging spectroscopy (FLIM) 359

11.2.2.1 Time-domain FLIM 359

11.2.2.2 Frequency-domain FLIM 361

11.2.2.3 Confocal FLIM (CFLIM) 362

11.2.2.4 Two-photon FLIM 362

11.3 Fluorescence correlation spectroscopy 364

11.3.1 Conceptual basis and instrumentation 364

11.3.2 Determination of translational diffusion coefficients 367

11.3.3 Chemical kinetic studies 368

11.3.4 Determination of rotational diffusion coefficients 371

11.4 Single-molecule fluorescence spectroscopy 372

11.4.2 Single-molecule detection in flowing solutions 372

11.4.3 Single-molecule detection using advanced fluorescence microscopy techniques 374.

Notes:

Includes bibliographical references and index.

Local Notes:

Acquired for the Penn Libraries with assistance from the Alumni and Friends Memorial Book Fund.

ISBN:

352729919X

OCLC:

45541665