Molecular Fluorescence
.pdfContents xi
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 di usion coe cients |
367 |
|||
11.3.3 |
Chemical kinetic studies |
368 |
|
|
|
11.3.4 |
Determination of rotational di usion coe cients |
371 |
|||
11.4 |
Single-molecule fluorescence spectroscopy 372 |
|
|||
11.4.1 |
General remarks |
372 |
|
|
|
11.4.2 |
Single-molecule detection in flowing solutions 372 |
11.4.3Single-molecule detection using advanced fluorescence microscopy
techniques 374
11.5Bibliography 378
Epilogue 381
Index 383
Molecular Fluorescence: Principles and Applications. Bernard Valeur
> 2001 Wiley-VCH Verlag GmbH
ISBNs: 3-527-29919-X (Hardcover); 3-527-60024-8 (Electronic)
xii
Preface
This book is intended for students and researchers wishing to gain a deeper understanding of molecular fluorescence, with particular reference to applications in physical, chemical, material, biological and medical sciences.
Fluorescence was first used as an analytical tool to determine the concentrations of various species, either neutral or ionic. When the analyte is fluorescent, direct determination is possible; otherwise, a variety of indirect methods using derivatization, formation of a fluorescent complex or fluorescence quenching have been developed. Fluorescence sensing is the method of choice for the detection of analytes with a very high sensitivity, and often has an outstanding selectivity thanks to specially designed fluorescent molecular sensors. For example, clinical diagnosis based on fluorescence has been the object of extensive development, especially with regard to the design of optodes, i.e. chemical sensors and biosensors based on optical fibers coupled with fluorescent probes (e.g. for measurement of pH, pO2, pCO2, potassium, etc. in blood).
Fluorescence is also a powerful tool for investigating the structure and dynamics of matter or living systems at a molecular or supramolecular level. Polymers, solutions of surfactants, solid surfaces, biological membranes, proteins, nucleic acids and living cells are well-known examples of systems in which estimates of local parameters such as polarity, fluidity, order, molecular mobility and electrical potential is possible by means of fluorescent molecules playing the role of probes. The latter can be intrinsic or introduced on purpose. The high sensitivity of fluorimetric methods in conjunction with the specificity of the response of probes to their microenvironment contribute towards the success of this approach. Another factor is the ability of probes to provide information on dynamics of fast phenomena and/or the structural parameters of the system under study.
Progress in instrumentation has considerably improved the sensitivity of fluorescence detection. Advanced fluorescence microscopy techniques allow detection at single molecule level, which opens up new opportunities for the development of fluorescence-based methods or assays in material sciences, biotechnology and in the pharmaceutical industry.
The aim of this book is to give readers an overview of molecular fluorescence, allowing them to understand the fundamental phenomena and the basic techniques, which is a prerequisite for its practical use. The parameters that may a ect the
Preface xiii
characteristics of fluorescence emission are numerous. This is a source of richness but also of complexity. The literature is teeming with examples of erroneous interpretations, due to a lack of knowledge of the basic principles. The reader’s attention will be drawn to the many possible pitfalls.
Chapter 1 is an introduction to the field of molecular fluorescence, starting with a short history of fluorescence. In Chapter 2, the various aspects of light absorption (electronic transitions, UV–visible spectrophotometry) are reviewed.
Chapter 3 is devoted to the characteristics of fluorescence emission. Special attention is paid to the di erent ways of de-excitation of an excited molecule, with emphasis on the time-scales relevant to the photophysical processes – but without considering, at this stage, the possible interactions with other molecules in the excited state. Then, the characteristics of fluorescence (fluorescence quantum yield, lifetime, emission and excitation spectra, Stokes shift) are defined.
The e ects of photophysical intermolecular processes on fluorescence emission are described in Chapter 4, which starts with an overview of the de-excitation processes leading to fluorescence quenching of excited molecules. The main excitedstate processes are then presented: electron transfer, excimer formation or exciplex formation, proton transfer and energy transfer.
Fluorescence polarization is the subject of Chapter 5. Factors a ecting the polarization of fluorescence are described and it is shown how the measurement of emission anisotropy can provide information on fluidity and order parameters.
Chapter 6 deals with fluorescence techniques, with the aim of helping the reader to understand the operating principles of the instrumental set-up he or she utilizes, now or in the future. The section devoted to the sophisticated time-resolved techniques will allow readers to know what they can expect from these techniques, even if they do not yet utilize them. Dialogue with experts in the field, in the course of a collaboration for instance, will be made easier.
The e ect of solvent polarity on the emission of fluorescence is examined in Chapter 7, together with the use of fluorescent probes to estimate the polarity of a microenvironment.
Chapter 8 shows how parameters like fluidity, order parameters and molecular mobility can be locally evaluated by means of fluorescent probes.
Chapter 9 is devoted to resonance energy transfer and its applications in the cases of donor–acceptor pairs, assemblies of donor and acceptor, and assemblies of like fluorophores. In particular, the use of resonance energy transfer as a ‘spectroscopic ruler’, i.e. for the estimation of distances and distance distributions, is presented.
In Chapter 10, fluorescent pH indicators and fluorescent molecular sensors for cations, anions and neutral molecules are described, with an emphasis on design principles in regard to selectivity.
Finally, in Chapter 11 some advanced techniques are briefly described: fluorescence up-conversion, fluorescence microscopy (confocal excitation, two-photon excitation, near-field optics, fluorescence lifetime imaging), fluorescence correlation spectroscopy, and single-molecule fluorescence spectroscopy.
This book is by no means intended to be exhaustive and it should rather be
xiv Preface
considered as a textbook. Consequently, the bibliography at the end of each chapter has been restricted to a few leading papers, reviews and books in which the readers will find specific references relevant to their subjects of interest.
Fluorescence is presented in this book from the point of view of a physical chemist, with emphasis on the understanding of physical and chemical concepts. E orts have been made to make this book easily readable by researchers and students from any scientific community. For this purpose, the mathematical developments have been limited to what is strictly necessary for understanding the basic phenomena. Further developments can be found in accompanying boxes for aspects of major conceptual interest. The main equations are framed so that, in a first reading, the intermediate steps can be skipped. The aim of the boxes is also to show illustrations chosen from a variety of fields. Thanks to such a presentation, it is hoped that this book will favor the relationship between various scientific communities, in particular those that are relevant to physicochemical sciences and life sciences.
I am extremely grateful to Professors Elisabeth Bardez and Mario Nuno BerberanSantos for their very helpful suggestions and constant encouragement. Their critical reading of most chapters of the manuscript was invaluable. The list of colleagues and friends who should be gratefully acknowledged for their advice and encouragement would be too long, and I am afraid I would forget some of them. Special thanks are due to my son, Eric Valeur, for his help in the preparation of the figures and for enjoyable discussions. I wish also to thank Professor Philip Stephens for his help in the translation of French quotations.
Finally, I will never forget that my first steps in fluorescence spectroscopy were guided by Professor Lucien Monnerie; our friendly collaboration for many years was very fruitful. I also learned much from Professor Gregorio Weber during a one-year stay in his laboratory as a postdoctoral fellow; during this wonderful experience, I met outstanding scientists and friends like Dave Jameson, Bill Mantulin, Enrico Gratton and many others. It is a privilege for me to belong to Weber’s ‘family’.
Paris, May 2001 |
Bernard Valeur |
Molecular Fluorescence: Principles and Applications. Bernard Valeur
> 2001 Wiley-VCH Verlag GmbH
ISBNs: 3-527-29919-X (Hardcover); 3-527-60024-8 (Electronic)
383
Index
a
acridine 59, 101, 319f |
||||
ACRYLODAN |
|
214 |
|
|
a nity constant |
339 |
|
||
4-amino-9-fluorenone |
213 |
|||
4-aminophthalimide |
207, 213, 219 |
|||
2-aminopyridine |
160 |
|
||
2-anilinonaphthalene |
218 |
|||
ANS 188, 214 |
|
|
||
anthracene |
54, 99, 160, 190 |
|||
anthracene-9-carboxylate 101 |
||||
anthracene-9-carboxylic acid 55, 77 |
||||
9-cyanoanthracene |
190 |
|||
9,10-diphenylanthracene 160, 190 |
||||
anthrone |
57 |
|
|
|
aromatic hydrocarbons 47, 54 |
||||
substituted |
56 |
|
||
association constant |
339 |
|||
auramine O |
66, 230f |
|
b
BCECF (biscarboxyethylcarboxyfluorescein)
280, 282, 284 |
|
|
|
Beer-Lambert Law |
23 |
|
|
benz(a)anthracene |
69 |
|
|
benzo[c]xanthene dyes |
279, 284 |
||
benzophenone |
57 |
|
|
benzoxazinone |
55, 60 |
|
|
bianthryl 65 |
|
|
|
binding constant |
|
339 |
|
biological membranes |
12, 237, 243, 262, 271 |
||
broadening |
|
|
|
homogeneous |
|
67 |
|
inhomogeneous |
67, 201, 265 |
||
Brownian motion, rotational 140 |
c
calcium crimson 296 calcium green 296
calcium orange 296 |
|
|
||
carbazole |
60, 77 |
|
|
|
CF (carboxyfluorescein) |
283 |
|||
charge transfer, intramolecular 62, 65, 206, |
||||
353 |
|
|
|
|
PCT (photoinduced charge transfer) cation |
||||
sensors |
298 |
|
|
|
TICT (twisted intramolecular charge |
||||
transfer) |
63, 64, 65, 206, 215, 230, 300, |
|||
302, 326 |
|
|
|
|
chromosome mapping |
|
358 |
||
CNF (carboxynaphthofluorescein) 280, 282 |
||||
Collins-Kimball’s theory |
80 |
|||
cooperativity |
345 |
|
|
|
correlation time, rotational 147, 241 |
||||
coumarin |
153, 213, 283 |
|||
7-alkoxycoumarins |
221 |
|||
7-aminocoumarins |
60 |
|||
4-amino-7-methylcoumarin 204 |
||||
cresyl violet |
160 |
|
|
|
crystal violet |
230f |
|
|
|
cyclodextrins |
265, 267, 323 |
d
DANCA |
214 |
|
|
dansyl |
325 |
|
|
DCM |
213 |
|
|
delayed fluorescence |
41 |
||
DENS |
77 |
|
|
di usion coe cient |
79 |
||
rotation |
146, 227, 230, 241 |
||
translation 227, 229, 234 |
|||
di usion-controlled reactions 80 |
|||
diphenylanthracene |
160, 190 |
||
diphenylhexatriene |
15, 240 |
||
diphenylmethane dyes 230 |
|||
dissociation constant |
339 |
DMABN (dimethylaminobenzonitrile) 63,
64, 213, 215, 217, 230f
384 |
Index |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
DNA |
12, 375, 376 |
|
|
|
|
|
erythrosin |
62, 190 |
|
|
|
|
||||||
|
|
|
|
|
|
|
|
|
|
||||||||||
|
dual-wavelength measurements 279, 338, 344 |
excimer |
73, 75, 94 |
|
|
|
|
||||||||||||
|
|
|
|
|
|
|
|
|
|
|
based cation sensors |
308 |
|
|
|||||
|
e |
|
|
|
|
|
|
|
|
|
intermolecular |
232, 234f |
|
|
|||||
|
Einstein coe cients |
|
28 |
|
|
|
intramolecular |
235f, 238 |
|
|
|||||||||
|
electron transfer |
|
73, 75, 90, 91, 279, 286, |
exciplex |
73, 94 , 99, 212 |
|
|
||||||||||||
292, 353 |
|
|
|
|
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|
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|
|
|
|
|
|||
|
electronic transitions |
20 |
|
|
f |
|
|
|
|
|
|
|
|
||||||
|
emission anisotropy |
|
125 , 227, 237, 243 |
fluidity |
226, 237 |
|
|
|
|
|
|||||||||
|
additivity law |
|
132 |
|
|
|
|
fluorenone |
57 |
|
|
|
|
|
|||||
|
anisotropic rotations |
147 |
|
fluorescein |
62, 77, 279f, 282, 283, 285 |
||||||||||||||
|
applications |
151 |
|
|
|
|
|
fluorescence |
15 |
|
|
|
|
|
|||||
|
asymmetric rotors |
148 |
|
|
analytical techniques |
15 |
|
|
|||||||||||
|
ellipsoids |
148 |
|
|
|
|
|
characteristics |
34 |
|
|
|
|
||||||
|
excitation polarization spectrum |
139, 141, |
e ects of hydrogen bonding 218 |
||||||||||||||||
166 |
|
|
|
|
|
|
|
|
e ects of intermolecular photophysical |
||||||||||
|
fundamental anisotropy |
136 |
|
processes |
72 |
|
|
|
|
||||||||||
|
G factor |
165 |
|
|
|
|
|
|
|
e ects of molecular structure |
54 |
||||||||
|
hindered rotations |
150, 152, 242 |
e ects of polarity |
200 |
|
|
|||||||||||||
|
isotropic rotations |
|
145, 240 |
|
e ects of specific interactions |
217 |
|||||||||||||
|
limiting anisotropy |
136 |
|
|
environmental e ects |
67 |
|
|
|||||||||||
|
measurement |
|
165 |
|
|
|
inner filter e ects |
161 |
|
|
|||||||||
|
polarization ratio |
|
130 |
|
|
lifetime |
190 |
|
|
|
|
|
|||||||
|
time-resolved |
|
189 |
|
|
|
|
polarization e ects |
163 |
|
|
||||||||
|
wobble-in-cone model |
150, 242 |
|
quantum yield |
46f, 53, 109, 159, 161 |
||||||||||||||
|
energy transfer |
|
73, 109, 359 |
|
spectra |
50, 53 |
|
|
|
|
|
||||||||
|
applications |
268 |
|
|
|
|
|
supersonic jets |
70 |
|
|
|
|||||||
|
coulombic mechanism 117 |
|
temperature e ects |
48 |
|
|
|||||||||||||
|
determination of distance |
249 |
|
fluorescence correlation spectroscopy (FCS) |
|||||||||||||||
|
Dexter theory |
|
122 |
|
|
|
|
364 , 375 |
|
|
|
|
|
||||||
|
di usion e ects |
257 |
|
|
|
chemical kinetic studies |
368 |
|
|||||||||||
|
dipole-dipole mechanism |
119, 122, 257 |
rotational di usion coe cients |
371 |
|||||||||||||||
|
distributions of distances |
254 |
|
single-molecule detection |
375 |
|
|||||||||||||
|
e ect of viscosity |
256 |
|
|
translational di usion coe cients 367 |
||||||||||||||
|
e ects of dimensionality |
260 |
|
fluorescence microscopy |
17, 353 |
||||||||||||||
|
e ects of restricted geometries |
261 |
clinical imaging |
359 |
|
|
|
||||||||||||
|
energy migration |
|
264 |
|
|
confocal |
354 |
|
|
|
|
|
|||||||
|
exchange mechanism |
117, 122f, 257 |
DNA sequencing |
359 |
|
|
|||||||||||||
|
excitation transport |
264 |
|
|
fluorescence lifetime imaging (FLIM) 359 |
||||||||||||||
|
Fo¨rster radius |
|
120, 248 |
|
|
NSOM |
|
357f |
|
|
|
|
|
||||||
|
Fo¨rster theory |
|
119 |
|
|
|
single-molecule detection |
374 |
|
||||||||||
|
non-radiative |
|
75, 82, 110, 113 , 247 |
single-photon timing |
359 |
|
|||||||||||||
|
orientational factor |
121, 249 |
|
two-photon excitation |
355, 358, 375 |
||||||||||||||
|
radiative |
110, 112 , 162 |
|
|
fluorescence polarization see emission |
||||||||||||||
|
rapid di usion limit |
259 |
|
|
anisotropy |
|
|
|
|
|
|||||||||
|
red-edge e ect |
265 |
|
|
|
fluorescence standards |
159f |
|
|||||||||||
|
selection rules |
122 |
|
|
|
fluorescence up-conversion |
210, 351 |
||||||||||||
|
spectroscopic ruler |
249 |
|
|
fluorescent pH indicators 276 , 280, 282f, |
||||||||||||||
|
strong coupling |
117 |
|
|
|
336 |
|
|
|
|
|
|
|
|
|||||
|
transfer e ciency |
|
121 |
|
|
fluorescent probes |
11 |
|
|
|
|||||||||
|
very weak coupling |
119 |
|
|
choice |
|
16 |
|
|
|
|
|
|
||||||
|
weak coupling |
119 |
|
|
|
microviscosity |
226 |
|
|
|
|||||||||
|
eosin |
62, 77, 279 |
|
|
|
|
|
pH see fluorescent pH indicators |
|||||||||||
|
equilibrium constants |
339 |
|
|
polarity |
200, 213 |
|
|
|
|
Index 385
fluorescent sensors |
273 |
fluoroimmunoassay |
12, 153 |
|
|||||||||
Agþ |
302 |
|
|
|
fluoroionophore |
275, 288, 290, 292 |
|||||||
anions |
315 |
|
|
fluorometry, steady-state |
155 |
|
|||||||
ATP |
319 |
|
|
|
fluorometry, time-resolved |
167 |
|
||||||
Ba2þ |
298, 307, 310, 312 |
color e ect |
180 |
|
|
|
|||||||
benzene |
327 |
|
|
comparison between pulse and phase |
|||||||||
boronic acid-based |
329 |
fluorometries |
195 |
|
|
||||||||
Br |
315 |
|
|
|
data analysis |
181 |
|
|
|
||||
Ca2þ |
295, 299, 302, 307, 312 |
deconvolution |
181 |
|
|
||||||||
carboxylates |
319f, 321 |
e ect of light scattering |
181 |
|
|||||||||
cations 287 , 336 |
global analysis |
184 |
|
|
|||||||||
chemical sensing devices 333 |
lifetime distributions 185, 187 |
|
|||||||||||
Cd2þ |
298 |
|
|
|
lifetime standards |
186 |
|
|
|||||
Cl |
315, 320 |
|
|
magic angle |
198 |
|
|
|
|||||
citrate |
323 |
|
|
|
maximum entropy method 187 |
||||||||
CuII |
293f |
|
|
|
phase-modulation fluorometry |
168, 177, |
|||||||
cyclodextrin-based |
323 |
180, 182, 192, 194 |
|
|
|||||||||
diols |
330 |
|
|
|
polarization e ects |
181, 196 |
|
||||||
F |
317 |
|
|
|
POPOP |
186 |
|
|
|
|
|||
Fura-2 |
304 |
|
|
|
pulse fluorometry |
167, 173 , 180f, 189, |
|||||||
gas |
336 |
|
|
|
194 |
|
|
|
|
|
|
||
glucose |
331 |
|
|
reduced chi square |
182 |
|
|||||||
H2PO4 320 |
|
|
single-photon timing technique |
173 |
|||||||||
hydroquinones |
333 |
stroboscopic technique |
176 |
|
|||||||||
Indo-1 |
304 |
|
|
time-correlated single-photon counting |
|||||||||
I 315 |
|
|
|
|
(TCSPC) |
173 |
|
|
|
||||
Kþ |
293, 298, 302, 305, 307, 312 |
weighted residuals |
183 |
|
|||||||||
Liþ |
299, 302, 312 |
Fo¨rster cycle |
103 |
|
|
|
|
||||||
Mag-Fura2 |
304 |
|
Fo¨rster theory |
247 |
|
|
|
||||||
Mag-indo1 |
304 |
|
Franck-Condon principle |
31 |
|
||||||||
main classes |
275 |
|
free volume |
228, 230, 232, 234, 236, 238 |
|||||||||
Mg2þ 295, 302, 307, 312 |
friction coe cient |
227, 229 |
|
||||||||||
Naþ |
295, 304, 307, 309, 313 |
Fura-2 |
302 |
|
|
|
|
|
|
||||
neutral molecules |
322f |
|
|
|
|
|
|
|
|
||||
NiII |
294 |
|
|
|
g |
|
|
|
|
|
|
|
|
nucleotides |
319f |
|
glass transition |
236, 238 |
|
|
|||||||
optode |
333 |
|
|
|
|
|
|
|
|
|
|
||
oxyquinoline-based cation sensors 310 |
h |
|
|
|
|
|
|
|
|||||
Pb2þ |
298 |
|
|
|
Ham e ect |
222 |
|
|
|
|
|||
PET (photoinduced electron transfer) cation |
heavy atom e ect |
|
|
|
|
||||||||
sensors |
292 |
|
intermolecular |
73 |
|
|
|||||||
pH see fluorescent pH indicators |
internal |
56 |
|
|
|
|
|
||||||
phosphates |
319f |
|
heterocyclic compounds |
59 |
|
||||||||
porphyrin-based |
329 |
hydrogen bonds |
59 |
|
|
|
|||||||
pyrophosphate ions 318 |
hydroxycoumarins |
60, 279 |
|
||||||||||
quinones 332 |
|
8-hydroxyquinoline |
59 |
|
|
||||||||
saccharide |
329f |
|
|
|
|
|
|
|
|
|
|||
SCN |
315 |
|
|
|
i |
|
|
|
|
|
|
|
|
Sr2þ |
310 |
|
|
|
immunoassays |
271 |
|
|
|
||||
steroid molecules |
322 |
Indo-1 |
302 |
|
|
|
|
|
|
||||
sulfonates |
320 |
|
indole |
60, 141 |
|
|
|
|
|
||||
surfactants |
322, 328 |
interactions |
|
|
|
|
|
|
|||||
uronic and sialic acids 321 |
dielectric |
201 |
|
|
|
|
|||||||
ZnII |
293, 296 |
|
hydrogen bonding |
202 |
|
386Index
Interactions (cont.)
solute-solvent |
202 |
|
Van der Waals |
201 |
|
internal conversion |
37 |
|
rate constant |
42 |
|
intersystem crossing |
38, 41 |
|
quantum yield |
46 |
|
rate constant |
42 |
|
j
Job’s method 346
l
Langmuir-Blodgett films 358 lasers
mode-locked 175 Ti:sapphire 175
LAURDAN 214 lifetime 47
amplitude-averaged 173 excited-state 42 intensity-averaged 172 radiative 44
triplet state 46 lipid bilayers 245
Lippert-Mataga equation 211 liquid crystals 358
living cells 12 luminescence 3
m
Mag-Fura2 |
303 |
|
|
||
Mag-indo1 |
303 |
|
|
||
malachite green |
66 |
|
|||
Marcus theory |
93 |
|
|
||
matrix isolation spectroscopy |
69 |
||||
6-methoxyquinoline |
101 |
|
|||
9-methylcarbazole |
190 |
|
|||
micelles |
87, 107, 188, 219, 237, 245, 369 |
||||
microviscosity |
226, 228, 242, 359 |
||||
modulator, electro optic 178 |
|
||||
molar absorption coe cient |
24, 27 |
||||
molecular mobility |
226, 235, 237 |
n
naphthacene 54 naphthalene 54, 56, 160 2-naphthol 101, 107
2-naphthol-6,8-disulfonate 101, 107
2-naphthol-6-sulfonate 101, 107
2-naphthylamine 101 NATA 190
near-field scanning optical microscopy
(NSOM) |
17, 356 |
nucleic acids |
12, 269, 271, 368, 375 |
o
orientation autocorrelation function 145 orientation polarizability 211
oscillator strength 24, 28 oxazines 62
p
parinaric acid |
240 |
|
|
||||
PBFI |
302 |
|
|
|
|
||
pentacene |
54 |
|
|
|
|
||
Perrin s equation |
146 |
|
|
||||
Perrin-Jablonski diagram |
34f |
||||||
perylene |
|
140 |
|
|
|
|
|
pH titrations |
337 |
|
|
|
|||
phenol |
|
101 |
|
|
|
|
|
phospholipid bilayers |
235 |
||||||
phospholipid vesicles |
242, 262 |
||||||
phosphorescence |
41 |
|
|
||||
quantum yield |
46 |
|
|
||||
photoluminescence |
4 |
|
|
||||
photomultipliers |
|
|
|
||||
microchannel plate |
175 |
||||||
polarity |
200 |
|
|
|
|
||
betain dyes |
202 |
|
|
||||
E T(30) scale |
203 |
|
|
||||
empirical scales |
202 |
|
|||||
p* scale |
204 |
|
|
|
|||
Py scale |
222 |
|
|
|
|||
polymers |
12, 232, 235, 245, 265, 270, 358 |
||||||
polynuclear complexes |
|
265 |
|||||
POPOP |
190 |
|
|
|
|
||
porous solids |
270 |
|
|
|
|||
PPO |
190 |
|
|
|
|
|
|
PRODAN |
77, 213f, 216 |
|
|||||
proteins |
12, 271, 358, 368, 375 |
||||||
proton transfer |
73, 75, 99 , 279, 353 |
||||||
pyranine |
|
14, 101, 108, 110, 278, 336 |
|||||
pyrene |
|
14, 95f, 222f, 234 |
|||||
pyrenebutyric acid |
77 |
|
|
||||
pyrenecarboxaldehyde |
221 |
||||||
pyrenehexadecanoic acid |
224 |
||||||
pyronines |
61 |
|
|
|
|
q
quantum counter 156, 158 quenchers 76
quenching of fluorescence 73 , 227, 232 dynamic 75, 77 , 86, 87, 90 heterogeneously emitting systems 89
|
|
|
|
|
|
|
|
Index |
387 |
|
|
|
|
|
|
||||
oxygen |
48 |
|
|
lifetime-based decomposition 194 |
|||||
Perrin s model |
84 |
|
pH dependence |
106, 110 |
|
||||
static |
75, 84 , 90 |
|
time-resolved |
192, 207 |
|
||||
quinine sulfate dihydrate |
160 |
spectrofluorometer |
156 |
|
|
||||
|
|
|
|
stability constant |
339 |
|
|
||
r |
|
|
|
Stern-Volmer kinetics |
77 |
|
|||
ratiometric measurements |
279; see also dual- |
Stern-Volmer relation |
78, 232 |
|
|||||
wavelengths measurements |
Strickler-Berg relation |
44 |
|
||||||
red-edge e ects |
67, 265, 267 |
stimulated emission |
39 |
|
|
||||
Rehm-Weller equation 92 |
|
Stokes-Einstein relation |
79, 147, 226, 228 |
|
|||||
rhodamines 61, 66 |
|
Stokes shift 38, 54, 211, 219 |
|||||||
rhodamine 101 |
66, 156, 160 |
streak camera |
176 |
|
|
|
|||
rhodamine B 61, 66, 190, 356 |
supramolecular systems |
270 |
|
||||||
rhodamine 6G |
55, 61, 378 |
surfactant solutions |
12 see also micelles |
||||||
rotors, molecular |
227, 230 |
|
|
|
|
|
|
||
rubrene |
190 |
|
|
t |
|
|
|
|
|
s
SBFI 302 selection rules 30
Shpol skii spectroscopy 69
single molecule fluorescence spectroscopy
17, 372 |
|
|
|
|
flowing solutions |
372 |
|
||
NSOM |
377 |
|
|
|
single-photon timing |
373 |
|||
site-selection spectroscopy 70 |
||||
Smoluchowski s theory |
80 |
|||
SNAFL |
279, 282, 284, 286, 336 |
|||
SNARF |
279, 282, 284, 286, 336 |
|||
solid matrices |
68 |
|
|
|
solid surfaces |
12 |
|
|
|
solvation dynamics |
209, 353 |
|||
solvatochromic shifts |
201, 202 |
|||
theory |
208 |
|
|
|
solvent relaxation 63, 206 |
SPA (sulfopropyl acridinium) 190, 316 spectra
correction |
158f |
emission |
48, 50, 157, 159 |
excitation |
48, 52, 157 |
p-terphenyl 190
TICT (twisted intramolecular charge transfer) see charge transfer
time-resolved fluorometry see fluorometry, time-resolved
TNS 214
transition moment 27 , 127, 134, 141
triphenylmethane dyes |
65, 230 |
|
triplet-triplet transition |
42 |
|
tryptophan 60, 160, 362 |
||
two-photon excitation 355 |
||
FLIM |
362 |
|
tyrosine |
77 |
|
u
UV-visible absorption spectroscopy 20 v
vesicles |
12, 219, 237, 245, 271 |
viscosity |
226, 230 |
w
Weber’s e ect 68 wobble-in-cone model 243
Molecular Fluorescence: Principles and Applications. Bernard Valeur
> 2001 Wiley-VCH Verlag GmbH
ISBNs: 3-527-29919-X (Hardcover); 3-527-60024-8 (Electronic)
1
Prologue
La lumie`re joue dans notre vie un roˆle essentiel: elle intervient dans la plupart de nos activite´s. Les Grecs de l’Antiquite´ le savaient bien de´ja`, eux qui pour dire ‘‘mourir’’ disaient ‘‘perdre la lumie`re’’.
Louis de Broglie, 1941
[Light plays an essential role in our lives: it is an integral part of the majority of our activities. The ancient Greeks, who for ‘‘to die’’ said ‘‘to lose the light’’, were already well aware of this.]