Principles of Fluorescence Spectroscopy

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932 INDEX Energy transfer [cont'd] theory of, 445–451 for donor–acceptor pair, 445–448 homotransfer and heterotransfer, 450–451 orientation factor, 448–449 transfer rate dependence on distance, overlap integral, and orientation factor, 449–450 Energy transfer, time-resolved, 477–501 acceptor decays, 489 advanced topics single-protein-molecule distance distribution, 496–497 biochemical applications of distance distributions, 490–496 calcium-induced changes in troponin C conformation, 490–493 DNA, four-way Holliday junction in, 493–494 hairpin ribozyme, 493 unfolding of yeast phosphoglycerate kinase, 494–495 diffusion effects for linked D–A pairs, 498–501 experimental measurement, 500–501 RET and diffusive motions in biopolymers, 501 simulations of RET for flexible pair, 499–500 distance distribution data analysis, 485–489 frequency-domain analysis, 485–487 time-domain analysis, 487 distance distribution in proteins analysis with frequency domain data, 485–487 distance distribution functions, 487 from time-domain measurements, 487 distance distributions from steady-state data, 443 D–A pairs with different R 0 values, 445–446 quenching, changing R 0 by, 447 distance distributions in glycopeptide, 495–496 distance distributions in peptides, 479–482 concentration of D–A pairs and, 482 cross-fitting data to exclude alternative models, 481–482 donor decay without acceptor, 482 rigid vs. flexible hexapeptide, 479–481 distance distributions in proteins, 482–485 domain motion in, 501 melittin, 483–485 incomplete labeling effects, 487–489 orientation factor effects, 489 representative literature references, 504 RET imaging, 497–498 Energy-transfer efficiency from enhanced fluorescence, 461–462 Energy-transfer immunoassay, 660–661 Energy-transfer stains and dyes, DNA, 715 Energy transfer to multiple acceptors in one, two, or three dimensions, 507–524 dimensionality effect on RET, 511–515 one dimension, 514–515 two dimensions, 512–514 with multiple acceptors, 515–516 aggregation of β-amyloid peptides, 515–516 RET imaging of fibronectin, 516 in presence of diffusion, 519–524 restricted geometries, 516–519 effect of excluded area, 518–519 RET in rapid-diffusion limit, 520–524 acceptor location in lipid vesicles, 521–522 retinal location in rhodopsin disk membranes, 522–524 three dimensions, 507–511 diffusion effects on RET with unlinked donors and acceptors, 508–509 experimental studies, 509–511 Enzyme cofactors intrinsic fluorescence, 63–65 time-domain lifetime measurement, 147 Enzyme kinetics, 770 Enzymes DNA, degradation, 808 turnover of, 780–781, 782 Eosin, 10, 633 Eosin-labeled ethanolamine, 448 Eosin-labeled lipid, 447 Epifluorescence, 41 Epi-illumination, 761 Erbium, 682, 683 Erythrosin, 408 Erythrosin B, 10 Escherichia coli maltose-binding protein, 653 Escherichia coli Tet repressor, 302–304 Estradiol, 80 Estrogens, intracellular RET indicator for, 458 1,2-Ethanedithiol (EDT), 86 Ethanol indole emission spectra in cyclohexane-ethanol mixture, 533–534 indole lifetime in, 533–534 polarizability properties, 207 rotational diffusion in, 367 specific solvent effects, 213, 214 tryptophan anisotropy spectra in, 533 Ethenoadenine derivatives, 287–288 Förster distances, 468 Ethers, 209, 279 Ethidium bromide, 75, 286 anisotropy decay, 402, 403–404, 432–433 DNA technology, 713, 714 energy transfer in one dimension, 514, 515 metal–ligand probes, 688 Ethidium homodimer, 75 Ethoxycoumarin, 230 Ethyl acetate, 206–207 Ethylaniline, 338 Ethylcellulose, 634 Ethylenediamine, dansylated, 229, 230 1-Ethylpyrene, 286 Europium, 3 as fluorophores, 679 Förster distances, 468 immunoassays, 659 metal-enhanced fluorescence, 843, 844 Excimers, 9, 269–270 DNA hybridization, 225, 718–719 Exciplex formation, 9, 278 Excitation, polarized, 778 Excitation anisotropy, two-photon and multiphoton, 21–22, 612–613 Excitation anisotropy spectra, 358–361, 531–533 Excitation filter, 41 Excitation photoselection of fluorophores, 357–358 Excitation spectra anisotropy, 358–360, 531–533 instrumentation, 27–31
PRINCIPLES OF FLUORESCENCE SPECTROSCOPY 933 corrected spectra, 51–52 distortions in, 30–31 ideal spectrofluorometer, 30 spectrofluorometer for high throughput, 29–30 polarization, tyrosine and tryptophan, 531–533 Excitation wavelength independence of fluorescence, 7–8 Excited charge-transfer complex (exciplex), 9, 278 Excited-state intramolecular photon transfer (ESIPT), 221–222 Excited-state reactions analysis by phase-modulation fluorometry, 265–270 apparent phase and modulation lifetimes, effects on, 266–267 wavelength-dependent phase and modulation values, 267–269 differential wavelength methods, 264 frequency-domain measurement of excimer formation, 196–197 naphthol dissociation, time-domain studies of, 264–265 naphthol ionization, 260–262 overview, 237–240 phase modulation, 265–270 phase-sensitive detection, 196–197 relaxation dynamics, 259–262 reversible two-state model, 262–264 steady-state fluorescence, 262–263 time-resolved decays for, 263–264 two-photon, 822–823 Exponential decays, frequency-domain lifetime measurements, 170–171 Extinction coefficients, 677 Extrinsic fluorophores, 15, 67 F FAD (flavin adenine dinucleotide), 64 imaging of, multiphoton microscopy, 617–618 quenching by adenine moiety, 278 time-domain lifetime measurement, 147–148 turnover of cholesterol oxidase, 780, 781 Fatty acid binding proteins, 226 Fatty acids. See also specific fatty acids fluorenyl, 295–296 membrane probes, 72 FCVJ, 225 Femtosecond titanium:sapphire lasers, 108–109 Ferredoxin, 584 Fibronectin, 516 Film polarizers, 51, 366 Films, 433–434 laser scanning TCSPC film, 748–750 surface-plasmon resonance, 863 wide-field frequency-domain, 746–747 Filters. See also Optical filters double dichroic, 824 holographic, 766 Rugate notch, 766 for single-molecule detection, 766–768 Fingerprint detection, 683 FITC. See Fluorescein isothiocyanate (FITC) Fitted function, 106 FKBP5901, 149, 592–593 Flash-EDT2 ,86 Flashlamps, 32 time-correlated single-photon counting, 112–114 Flavin adenine dinucleotide. See FAD Flavin mononucleotide, 16, 64, 316, 317, 397, 398 Flavoprotein, electron transfer, 315–316, 317 Flavoproteins, 65 Flexible D–A pairs, 499–500 Flexible hexapeptide, distance distributions, 479–481 Flow cytometry, 85 DNA fragment sizing, 715 Flow measurements using fluorescence correlation spectroscopy, 830–832 Fluid flow, 225 Fluorenyl fatty acids, 295–296, 299 Fluorenyl-PC, 72 Fluorescein, 2, 3, 20, 69, 70, 122, 695 anisotropic rotational diffusion, 428 brightness, 818 DNA energy-transfer reaction, 454 DNA technology, 707, 717, 719, 726 filters, 38 melittin labeled with, 374 metal–ligand complexes, 691 methionyl-tRNA synthetase labeled with, 370–371 as pH sensor, 637–639 quantum yield standards, 52 quenching, 310–312 resonance energy transfer, 365 time-resolved RET imaging, 498 Fluorescein isothiocyanate (FITC), 16, 17, 68 FITC-dextran, 635 Förster distances, 468 Fluorescence energy-transfer efficiency, 461–462 intensity fluctuations, 798–799, 800 metal-enhanced, 841 Fluorescence anisotropy. See Anisotropy Fluorescence correlation spectroscopy, 22–23, 757, 797–832 applications to bioaffinity reactions, 807–810 association of tubulin subunits, 807–808 DNA applications, 808–810 protein binding to chaperonin GroEL, 807 detection of conformational dynamics in macromolecules, 820–821 dual-color, 823–828 applications of, 826–828 DNA cleavage by restriction enzyme, 826 instrumentation for, 824 literature references, 837 theory of, 824–826 effects of chemical reactions, 816–817 examples of experiments, 805–807 effect of fluorophore concentration, 805–806 effect of molecular weight on diffusion coefficients, 806–807 flow measurements, 830–832 fluorescence intensity distribution analysis, 817–819 intersystem crossing, 815–816 theory for, 816 literature references, 837–839 in membranes, 810–815 binding to membrane-bound receptors, 813–815 lateral diffusion, 812–813 principles of, 798–799 rotational diffusion and photon antibunching, 828–830 theory of, 800–805 multiple diffusing species, 804–805 occupation numbers and volumes, 804 translational diffusion, 802–804 time-resolved, 819–820
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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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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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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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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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