Principles of Fluorescence Spectroscopy

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938 INDEX Internal charge transfer (ICT) state, 207, 220–221, 223–224, 229, 232 Internal conversion in fluorophores, 5 Intersystem crossing, 5–6, 334–335, 815–816 Intracellular protein folding, 454–455 Intramolecular quenching, 314–317 Intrinsic fluorophores, 15 Intrinsic/natural fluorescence, 15, 63–67 enzyme cofactors, 63–65 multiphoton excitation, 613–616 of proteins, 594–596 In-vivo imaging, sensors, 656–658 Iodide absorption spectra, 303 autocorrelation function in fluorescence correlation spectroscopy, 816 quenching by, 18, 278, 279, 287, 288, 290, 303–304, 306 chloride probe, 631 proteins, 290, 546–547 Iodine, 5 Iodoacetamides, 68 Ionic quenchers, 290. See also specific ions Ion indicators/probes/sensors. See also specific elements/ions analyte recognition probes, 644–650 calcium and magnesium, 647–650 cation probe specificity, 644 sodium and potassium, 645–647 chloride, 631–632 energy-transfer mechanisms, 635–636 oxygen, 627–630 photoinduced electron-transfer (PET) probes, 316, 318, 335–336, 627, 641–643 Ionization naphthol, 260–262 tyrosine, 534–535 IR-125, 74 IR-144, 174 Iridium metal–ligand complexes, 684 I7S, 539 Isomers, rotational, 253–255, 529, 578 Isoxanthopterin (IXP), 75, 76 Iterative reconvolution, 131 J Jablonski, Alexander, 4–5 Jablonski diagram, 3–6 collisional quenching and resonance energy transfer, 11 lanthanide probe, 87 metal-enhanced fluorescence, 842–843 red-edge excitation, 257 solvent effects, 207 solvent relaxation, 206, 238 JF9, 780 K Karyotyping, spectral, 730–732 Kasha's rule, 7, 359 Kerr lens mode locking, 109 ∆5-3-Ketosteroid isomerase (KSI), 557 KFU 111, 635, 636 Kinase, intracellular, 823 Kinesin, 784 Kinetics, literature references, 838 Klenow fragment of DNA polymerase, 417 Kretschmann configuration, 862 L Labeling, incomplete, energy-transfer studies, 452, 487–489 α-Lactalbumin, 806 Lactate dehydrogenase, 559–560, 584 Lactoglobulin, 368 LaJolla Blue, 719 Lamps. See Light sources Lanthanide MLCs, 87–88 long-lifetime probes, 87–88 quenching, 523 retinal energy-transfer study, 523 Lanthanides, 3, 467 fingerprint detection, 683 as fluorophores, 679–683 nanoparticles, 682 near-infrared emitting, 682, 683 resonance energy transfer, 680–681 as sensors, 681–682 Laplace method, 130 Laser diodes, 21 DNA technology, 711 frequency-domain lifetime measurements, 174 human serum albumin measurement with, 71, 72 instrumentation, 33, 34 time-domain lifetime measurements, 107–108 Laser line filter, 39 Lasers DNA technology, 711 dual-color fluorescence cross correlation spectroscopy, 824 frequency-domain lifetime measurements, 173–174 monochromators, 34 multiphoton excitation, 577 time-correlated single-photon counting film, 748–750 picosecond dye devices, 105–107 solid-state devices, 107 time-resolved protein fluorescence, 577 Laser scanning confocal microscopy (LSCM), 764 Laser scanning microscopy (LSM), 748–750 frequency-domain, 750–751 imaging of cellular biomolecules, 750 Lateral diffusion in membranes, 300–301, 812–813 Laurdan, 219, 220, 221 Least-squares analysis, 160, 487 frequency-domain lifetime measurements, 161–162 Stern-Volmer plots, 304 time-domain lifetime measurements, 129–130 TRES, 254 Least-squares fitting, 103 Lecithin:cholesterol acyltransferase, 454 Lecithin vesicles, 248 LED. See Light-emitting diodes Leucine, 294 L-format anisotropy measurement or single-channel method, 361–363 Lifetime-based probes and sensing, 626 DNA sequencing, 712 metal–ligand complexes, 88 (See also Metal–ligand complexes (MLCs)) oxygen, 628–629
PRINCIPLES OF FLUORESCENCE SPECTROSCOPY 939 Lifetime (decay time), 5–6 antibodies, 498 collisional quenching and, 256–257 estimation, single-molecule detection, 787–788 fluorescence-lifetime imaging microscopy, 741–752 frequency-domain (See Frequency-domain lifetime measurements) long-wavelength long-lifetime fluorophores, 695–697 meaning of, 99 metal-enhanced fluorescence, 842–843 metal–ligand complexes, 683–695 natural, 10, 537 phase-modulation apparent lifetimes, 191–192 phase angle and modulation, 192–194 principles, 9–12 quenching, 12 timescale of molecular processes in solution, 12 protein folding and, 558 quantum yield and, 537 solvent effects, 222–223 steady-state intensity and, 14 time-domain (See Time-domain lifetime measurements) Lifetime distributions, time-domain lifetime measurements, 143 Lifetime imaging, 743 Lifetime reference, 133 Lifetime-resolved anisotropy, 435–436 Lifetime-resolved emission spectra, relaxation dynamics, 255–257 Lifetime standards, 169 Lifetime-weighted quantum yield, 142 Ligand analogs, 80 Ligand-centered absorption, 687 Light absorption and deviation from Beer-Lambert law, 59 Light diffraction, 37 Light-emitting diodes, 33 frequency-domain lifetime measurements, 174–175, 189 instrumentation, 33–34 time-domain lifetime measurements, 108 Light-generated DNA arrays, 734 Light-generated probe arrays, DNA hybridization, 734 Light modulators frequency-domain lifetime measurements, 165–166 monochromators, 34–35 Light sources, 28 instrumentation, 31–34 lamps in corrected emission spectra, 34 time-correlated single-photon counting, 107–114 femtoscecond titanium:sapphire lasers, 108–109 flashlamps, 112–114 picosecond dye lasers, 110–112 synchrotron radiation, 114 Linear-focused dynode chain PMTs, 118 Linearity, time-to-amplitude converter (TAC), 115–116 Lipid probes, 690–691 Lipids energy transfer, 521–522 membrane (See Membranes) metal–ligand complex probes, 690 pyrenyl-labeled, 72 pyrenyl-PC, 72 Lipids, boundary lipid-water partitioning effects, 298–300 Lipids, membrane boundary, 298 distributions around Gramicidin, 463–465 energy transfer, 513 exchange of, 465 Lipobeads, 656 Lipocalin, 561, 563 Lippert equation, 208–213 application of, 212–213 derivation of, 210–212 Lippert-Mataga equation, 208–210 Lippert plots, solvent effects, 215–216 Liquid crystals, 377 Lissamine rhodamine B, 70 Liver alcohol dehydrogenase (LADH), 395, 584 Localized quenchers, membranes, 332 Locally excited state, 220–221 Longitudinal relaxation times, 250, 252 Long-lifetime probes, 86–88 Long-pass filters, 38, 39 Long-wavelength long-lifetime fluorophores, 695–697 Lorentzian distributions, 487 correlation time, 393 Lorentzian lifetime distributions, 143 Low-frequency polarization, 210 Low-pressure mercury lamps, 33 Lucigenin, 631 Lumazine, 589 Luminescence, defined, 1 Luminescence decays, 129 Lysozyme, 288 M MACA (N-methylacridinium-9-carboxamide), 631 Macromolecules, detection of conformational dynamics, 820–821 Magic-angle conditions, 43 Magic-angle polarizer conditions, 364 Mag-INDO-1, 646 Magnesium indicators/probes/sensors, 17, 187–188, 647–650 Magnesium Orange, 646, 649 Magnesium oxide scatterer, 53 Magnetic resonance imaging (MRI), 406, 742 Mag-Quin-2, 187–188 Malachite green, 514 Maleimides, 68 Maltose binding protein (MBP), 457–458, 653 Manganese, 278 Maximum entropy analysis (MEM), 185–186 Maximum entropy method, 148–149 Melittin, 20, 547, 548 anisotropy decay, 398, 453, 590–591 association reactions detection from homotransfer, 374 distance distributions, 483–485 frequency-domain lifetime measurements, 171, 188–189 phosphorescence, 599 RET distance measurements, 451–452 rotational correlation time, 590 Membrane binding site polarity, solvent effects on emission spectra, 206–207
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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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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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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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Page 929 and 930: Index A Absorption spectroscopy, 12
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