Principles of Fluorescence Spectroscopy

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948 INDEX Rehm-Weller equation, 337 Relaxation, 7, 12 Relaxation dynamics, 237–270 analysis of excited-state reactions, 265–270 continuous and two-state spectral relaxation, 237–239 phase modulation studies of solvent relaxation, 265–267 excited-state ionization of naphthol, 260–262 excited-state processes, overview, 237–240 excited-state reactions, 259–262 lifetime-resolved emission spectra, 255–257 multi-exponential spectral relaxation, measurement, 252–253 picosecond relaxation in solvents, 249–252 multi-exponential relaxation in water, 251–252 theory of time-dependent solvent relaxation, 250–251 red-edge excitation shifts, 257–259 energy transfer, 259 membranes, 258–259 solvent relaxation vs. rotational isomer formation, 253–255 time-resolved emission spectra (TRES) analysis, 246–248 anthroyloxy fatty acids, 248–249 labeled apomyoglobin, 243–244 measurement, 240–242 membranes, spectral relaxation in, 245–249 overview, 239–240 proteins, spectral relaxation in, 242–243 synthetic fluorescent amino acid, 244–245 time-resolved emission spectra (TRES) measurement direct recording, 240–241 from wavelength-dependent decays, 241–242 TRES vs. DAS, 255 Resonance, surface-plasmon, 861–865 Resonance energy transfer (RET), 13, 144, 375, 443. See also Energy transfer applications, 20–21, 490–496 characteristics of, 443–445 comparison with quenching, 331–334 distance dependence, 332–333 encounter complexes and quenching efficiency, 333–334 Dexter interaction, 335 and diffusive motions in biopolymers, 501 effects on anisotropy, 364–365 efficiency, 333 literature references, 839 molecular information from fluorescence, 19–20 principles, 13–14 sensors, 626 time-resolved RET imaging, 497–498 Restricted geometries, energy transfer, 516–519 Restriction enzymes, 712–713, 826 Restriction fragment length polymorphisms (RFLPs), 713 RET. See Resonance energy transfer (RET) Retinal, 522–524 Reversible two-state model, 262–264 differential wavelength methods, 264 steady-state fluorescence of, 262–263 time-resolved decays for, 263–264 Rhenium complex, 520 Rhenium metal–ligand complexes, 88, 684, 686 immunoassays, 692–693 spectral properties, 687 Rhod-2, 645 Rhodamine, 2, 3, 20, 314, 315 anisotropy decay, 394, 416 DNA technology, 723, 725, 729 glucose sensor, 635 quantum yield standards, 54 time-resolved RET imaging, 498 Rhodamine 800, 74 Rhodamine B, 51, 279 metal-enhanced fluorescence, 848 Rhodamine derivatives, 74, 76 Förster distances, 468 structures of, 74–75 Rhodamine 6G, 122, 514 concentration in fluorescence correlation spectroscopy, 805–806 single-molecule detection, 759–760 Rhodamine 6G dye laser. See R6G laser Rhodamine green (RhG), 824 Rhodamines, 68–69 Rhodopsin disk membranes, retinal in, 522–524 Riboflavin, 64 Ribonuclease A, 501, 558–559 Ribonuclease T i , 536, 548, 588–589 anisotropy decays, 584, 585 Ribozymes energy transfer, 460 hairpin, 493 representative literature references, 505 in single-molecule detection, 23–24, 775 Ribozyme substrate binding, 311–312 Rigid rotor, 397–398 Rigid vs. flexible hexapeptide, distance distributions, 479–481 RNA energy transfer, 459–461 imaging of intracellular RNA, 460–461 Room-temperature phosphorescence of proteins, 599 Rose bengal, 848 Rotamers (rotational isomers), 253–255, 529, 578–580 Rotational correlation time, 102–103, 353 ellipsoids, 423–425 Rotational diffusion, 12, 13, 102, 168, 365, 422–423, 828–830 anisotropy decay ellipsoids, theory, 425–426 frequency-domain studies of, 427–429 non-spherical molecules, 418–419 time-domain studies of, 426–427 membranes, hindered, 399–402 oxytocin, 399 Perrin equation, 366–370 rotational motions of proteins, 367–369 stick vs. slip rotational diffusion, 425 Rotational isomer formation, 253–255, 529, 578–580 Rotational motion measurement of, 102 single-molecule detection, 775–779 Rotors hindered, 391–392 rigid, 397–398 R6G laser, 111, 112, 120 fluorescence intensity distribution analysis, 818, 819 single-molecule detection, 777–778 R3809U, 117
PRINCIPLES OF FLUORESCENCE SPECTROSCOPY 949 Ru(bpy) 2 phe-C 12 , 189, 190 Rugate notch filters, 766 Ruthenium metal–ligand complexes, 88, 684, 686 S Sample geometry effects, 55–57 Sample preparation, common errors in, 57–58 SBFI (sodium-binding benzofuran isophthalate), 645–647 Scanning fluorescence correlation spectroscopy literature references, 839 Scattered light, 366, 766 phase-sensitive detection, 196, 197 Scattered light effect, frequency-domain lifetime measurements, 172–173 Second-order transmission, monochromator, 37 Segmental mobility, biopolymer-bound fluorophore, 392–393 Selex, 725 Self-quenching, 69–70 metal-enhanced fluorescence, 853–854 Semiconductor nanoparticles, 675–678 Seminaphthofluoresceins (SNAFLS), 640–641 Seminaphthorhodofluors (SNARFS), 640–641 Senile plaques, 498 Sensing and sensors, 78–79, 623–663 analyte recognition probes, 643–650 calcium and magnesium, 647–650 cation probe specificity, 644 intracellular zinc, 650 sodium and potassium, 645–647 theory of, 644–645 calcium, 784 cardiac, 662 chloride, 631–632 clinical chemistry, 623–624 by collisional quenching, 627–633 chloride, 631–632 miscellaneous, 632 oxygen, 627–630 energy transfer glucose, 634–635 ion, 635–636 pH and CO 2 , 633–634 theory for, 636–637 energy-transfer, 633–637 GFP sensors, 654–655 intrinsic, 655 using resonance energy transfer, 654–655 glucose-sensitive fluorophores, 650–651 immunoassays, 658–663 ELISA, 659 energy transfer, 660–661 fluorescence polarization, 661–663 time-resolved, 659–660 in-vivo imaging, 656–658 lanthanides, 659, 681–682 literature references, 795 mechanisms of sensing, 626–627 metal–ligand complexes (See Metal–ligand complexes (MLCs)) molecular information from fluorescence, 17–20 new approaches to, 655–656 pebble sensors and lipobeads, 655–656 pH, two-state sensors, 637–641 blood gases, optical detection of, 637 pH sensors, 637–641 phosphorescence, 629 photoinduced electron-transfer (PET), 316, 318 photoinduced electron-transfer (PET) probes for metal ions and anions, 641–643 probes, 79–81 proteins as sensors, 651–652 based on resonance energy transfer, 652–654 RET, 458–459 spectral observables, 624–626 lifetime-based sensing, 626 optical properties of tissues, 625 Serotonin, 615–616 Serum albumin, 13, 71, 72, 304, 584, 589, 678 anisotropy decay of, 413–414 intensity decay of, 583, 584 laser diode excitation, measurement with, 71, 72 metal-enhanced fluorescence, 851–853 rotational correlation time, 368 Shear stress on membrane viscosity, 225 β-Sheet structure, 561–562, 563 Side-window dynode chain PMTs, 118 Signaling, intracellular, 459 Signal-to-noise ratio in single-molecule detection, 784–786 Silicone, oxygen sensor support materials, 627, 628, 629 Silver, 278, 279 effect on resonance energy transfer, 854–855 Silver colloids, 845–846 Silver island films, 848 Simazine, 660 Simulated intensity decay, 101, 102 Single-channel anisotropy measurement method, 361–363 Single-exponential decay, 120, 121 spherical molecules, 367 time-dependent intensity, 198 Single-exponential decay law, 346 Single-exponential fit, FD intensity decay approximation, 479 Single-exponential intensity and anisotropy decay of ribonuclease T I , 584, 585 Single-molecule detection, 23–24, 757–788 advanced topics, 784–788 lifetime estimation, 787–788 polarization measurements and mobility of surface-bound fluorophores, 786–787 polarization of single immobilized fluorophores, 786 signal-to-noise ratio in single-molecule detection, 784–786 biochemical applications, 770–773, 780–784 chaperonin protein, 771–773 conformational dynamics of Holliday junction, 782–783 enzyme kinetics, 770 molecular beacons, 782 motions of molecular motors, 784 single-molecule ATPase activity, 770–771 single-molecule calcium sensor, 784 turnover of single enzyme molecules, 780–781, 782 detectability of single molecules, 759–761 instrumentation, 764–768 detectors, 765–766 optical filters, 766–768 internal reflection and confocal optics, 760–762 confocal detection optics, 761–762 total internal reflection, 760–761 literature references, 788, 791–795 optical configurations for, 762–764
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Principles of Fluorescence Spectros
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Joseph R. Lakowicz Center for Fluor
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Preface The first edition of Princi
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x GLOSSARY OF ACRONYMS DNS dansyl o
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xii GLOSSARY OF ACRONYMS TRES time-
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xiv GLOSSARY OF MATHEMATICAL TERMS
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xvi CONTENTS 2.9.4. Conversion betw
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xviii CONTENTS 6. Solvent and Envir
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xx CONTENTS 10.4.4. Alignment of Po
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xxii CONTENTS 14.7.1. Simulations o
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xxiv CONTENTS 19.12. New Approaches
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xxvi CONTENTS 25.3. Optical Propert
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2 INTRODUCTION TO FLUORESCENCE Figu
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6 INTRODUCTION TO FLUORESCENCE inst
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10 INTRODUCTION TO FLUORESCENCE Fig
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12 INTRODUCTION TO FLUORESCENCE ns,
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14 INTRODUCTION TO FLUORESCENCE 1.7
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24 INTRODUCTION TO FLUORESCENCE due
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28 INSTRUMENTATION FOR FLUORESCENCE
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3 Fluorescence probes represent the
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PRINCIPLES OF FLUORESCENCE SPECTROS
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98 TIME-DOMAIN LIFETIME MEASUREMENT
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100 TIME-DOMAIN LIFETIME MEASUREMEN
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5 In the preceding chapter we descr
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6 Solvent polarity and the local en
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238 DYNAMICS OF SOLVENT AND SPECTRA
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278 QUENCHING OF FLUORESCENCE 8.1.
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280 QUENCHING OF FLUORESCENCE Figur
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282 QUENCHING OF FLUORESCENCE ly ev
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290 QUENCHING OF FLUORESCENCE relat
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298 QUENCHING OF FLUORESCENCE σ ar
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320 QUENCHING OF FLUORESCENCE 47. R
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322 QUENCHING OF FLUORESCENCE 113.
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328 QUENCHING OF FLUORESCENCE P8.3.
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330 QUENCHING OF FLUORESCENCE P8.11
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332 MECHANISMS AND DYNAMICS OF FLUO
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10 Measurements of fluorescence ani
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444 ENERGY TRANSFER Figure 13.1. Fl
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446 ENERGY TRANSFER wavelength is e
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448 ENERGY TRANSFER Table 13.1. Cal
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450 ENERGY TRANSFER Figure 13.6. Ab
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452 ENERGY TRANSFER safe for many p
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454 ENERGY TRANSFER Figure 13.12. P
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456 ENERGY TRANSFER Figure 13.16. E
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458 ENERGY TRANSFER Figure 13.21. R
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460 ENERGY TRANSFER Figure 13.24. R
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462 ENERGY TRANSFER tiple acceptors
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464 ENERGY TRANSFER Figure 13.29. A
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466 ENERGY TRANSFER Figure 13.33. F
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468 ENERGY TRANSFER Table 13.3. Rep
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470 ENERGY TRANSFER 55. Clegg RM. 1
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472 ENERGY TRANSFER Tong AK, Jockus
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474 ENERGY TRANSFER Figure 13.39. O
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14 In the previous chapter we descr
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16 The biochemical applications of
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578 TIME-RESOLVED PROTEIN FLUORESCE
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608 MULTIPHOTON EXCITATION AND MICR
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19 Fluorescence sensing of chemical
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676 NOVEL FLUOROPHORES Figure 20.1.
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678 NOVEL FLUOROPHORES Figure 20.7.
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680 NOVEL FLUOROPHORES Figure 20.12
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698 NOVEL FLUOROPHORES 33. Acherman
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700 NOVEL FLUOROPHORES 103. Demas J
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702 NOVEL FLUOROPHORES 176. Montalt
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21 During the past 20 years there h
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22 Fluorescence microscopy is one o
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758 SINGLE-MOLECULE DETECTION Figur
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766 SINGLE-MOLECULE DETECTION small
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770 SINGLE-MOLECULE DETECTION Table
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786 SINGLE-MOLECULE DETECTION face
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788 SINGLE-MOLECULE DETECTION TCSPC
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790 SINGLE-MOLECULE DETECTION 53. H
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792 SINGLE-MOLECULE DETECTION Ke PC
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794 SINGLE-MOLECULE DETECTION Polar
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862 RADIATIVE-DECAY ENGINEERING: SU
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Appendix I Corrected Emission Spect
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Appendix II Fluorescence Lifetime S
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890 APPENDIX III P ADDITIONAL READI
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892 APPENDIX III P ADDITIONAL READI
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894 ANSWERS TO PROBLEMS A1.4. A. Th
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