Principles of Fluorescence Spectroscopy

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924 INDEX Anisotropy, 353–378 applications, 20–21 biochemical applications, 372–374 helicase-catalyzed DNA unwinding, 373–374 melittin association detected from homotransfer, 374 peptide binding to calmodulin, 372–373 trp repressor binding to DNA, 373 definition, 353–355 electronic state resolution from polarization spectra, 360–361 emission wavelength and, 437–438 energy transfer, 453 excitation anisotropy spectra, 358–361, 531–533 excitation photoselection of fluorophores, 357–358 instrumentation, 36–37 lifetime-resolved, 435–436 measurement, 361–366 comparison of methods, 363–364 depolarization, trivial causes, 365–366 factors affecting anisotropy, 366 L-format or single channel method, 361–363 magic-angle polarizer conditions, 364 polarizer alignment, 364 resonance energy-transfer effects, 364–365 T-format or two-channel method, 363–364 total intensity, 364 membranes and membrane-bound proteins, 374–377 distribution, 375–377 membrane microviscosity, 374–375 metal–ligand complexes, 685, 688–689 multiphoton excitation, 612–613, 615 excitation photoselection for two-photon excitation, 612 two-photon anisotropy for DPH, 612–613 Perrin equation, rotational diffusion effects, 366–370 examples of Perrin plots, 369–370 rotational motions of proteins, 367–369 Perrin plots of proteins, 370–372 segmental motion effects, 436 photoselection, 357–358 polarization, molecular information from fluorescence, 19 principles, 12–13 proteins association reactions, 372–374 Perrin plots, 370–372 rotational motion, 102–103 segmented motions, 436 single-molecule detection, 775–776 Soleillet's rule, depolarization factor multiplication, 436–437 theory, 355–358 transition moments, 377–378 tryptophan, 531–532 Anisotropy, advanced concepts, 413–438 associated anisotropy decay, 413–417 theory, 414–415 time-domain measurements, 415–417 biochemical examples, 417–419 frequency-domain measurements, 417–418 time-domain studies of DNA binding to Klenow fragment, 417 DNA, 432–433 ellipsoids, anisotropy decay, 419–420 ellipsoids of revolution, 420–425 oblate, intuitive description of, 422–423 rotational correction times, 423–425 simplified, 421–422 stick vs. slip rotational diffusion, 425 global anisotropy decay analysis, 429–432 with collisional quenching, 430–431 with multi-wavelength excitation, 429–430 quenching application to protein anisotropy decays, 431–432 planar fluorophores with high symmetry, 435 rotational diffusion of ellipsoids, theory, 425–426 frequency-domain studies of, 427–429 of non-spherical molecules, 418–419 time-domain studies, 426–427 transition moments, 433–435 Anisotropy decay laws, 390–394 associated delays, 393 correlation time distributions, 393 hindered rotors, 391–392 non-spherical fluorophores, 391 segmental mobility of biopolymer-bound fluorophore, 392–393 Anisotropy decays, 14, 102–103, 413–417 energy transfer, 453 in frequency domain, 588–589 melittin, 590–591 proteins, 587–588, 589–591 ribonuclease T 1 , 584–585 Anisotropy decays, time-dependent, 383–409 analysis, 387–389 frequency-domain decays, 390 r 0 value, 389 time-domain decays, 387–389 correlation time imaging, 406–408 laws governing, 390–394 associated decays, 393 correlation time distributions, 393 decays of rhodamine green and rhodamine green-dextran, 394 hindered rotors, 391–392 non-spherical fluorophores, 391 segmental mobility of biopolymer-bound fluorophore, 392–393 membrane probe, characterization of, 401–402 membranes, hindered rotational diffusion in, 399–402 microsecond, 408–409 long-lifetime metal–ligand complexes, 408–409 phosphorescence, 408 nucleic acids, 402–406 DNA binding to HIV integrase, 404–406 DNA oligomer hydrodynamics, 403 intracellular DNA dynamics, 403–404 proteins, frequency-domain decays of, 383–387, 397–399 apomyoglobin, rigid rotor, 397–398 melittin, 398 oxytocin, picosecond rotational diffusion, 399 proteins, time-domain decays of, 383–387, 394–397 alcohol dehydrogenase, 395 domain motions of immunoglobulins, 396–397 free probe, effects of, 397 phospholipase A 2 , 395 Subtilisin Carlsberg, 395–396 Anisotropy fluorescence correlation spectroscopy, 829 Anisotropy polarization, 624–625 molecular information from fluorescence, 19 Anisotropy spectra, excitation, 358–361, 531–533 Annexins, 148 Annexin V, 148, 585–587
PRINCIPLES OF FLUORESCENCE SPECTROSCOPY 925 ANS time-resolved fluorescence correlation spectroscopy, 819 8,1-ANS (8-anilino-1-naphthalenesulfonate), 226–227, 258 ANS (1-anilinonaphthalene-6-sulfonic acid), 71–72 ANS (1-anilinonaphthalene-8-sulfonic acid) anisotropy decay, 417–418 red-edge excitation, 258 solvent effects, 212 6-ANS (6-anilinonaphthalene-2-sulfonic acid), 230 Förster distances, 468 ANS derivatives, solvent effects, 212 Anthracene, 3, 56, 59, 316, 632 emission spectrum, 9, 199 frequency-domain lifetime measurements, 170–171 orientation factor, 465 quenchers of, 278, 279 TRES and DAS comparison, 255 Anthranilic acid frequency-domain lifetime measurements, 180–185 mixture lifetime resolution, 138–141 time-domain lifetime measurements, 137–138 9-Anthroxycholine (9-AC), 226–227 Anthroyl fatty acid, 72 Anthroyloxy fatty acids, 279 9-AS, 359–360 12-AS, 360, 375 Förster distances, 468 solvent relaxation, 248–249 TRES, 248–249 12-Anthroyloxy stearate, 375 9-Anthroyloxy stearic acid, 359–360, 369 Anthroylxanthyl cations, quenchers of, 279 Antibody labeling, 69 APC (allophycocyanin) Förster distances, 468 Apoazurin, 546–547, 583–584 Apo E3, 454–455 Apo E4, 454–455 Apolipoproteins, 454 Apomyoglobin, 17, 368, 417–418, 549 anisotropy decay, 397–398 protein folding, 560 time-resolved emission spectra (TRES), 243–244 Apoperoxidase, 368 Apparent distance distributions, 478–480 Apparent phase and modulation lifetimes, 191–192, 266–267 Aptamers, 724–726 APTRA, 644, 649 Archaebacterial histone-like protein (HTa), 544–545 Arc lamps, 28, 31, 33 Argon-hydrogen flashlamps, 113–114 Argon-ion lasers, 39, 109, 110–111, 715–716 Aromatic amino acids, 314. See also Phenylalanine; Tryptophan; Tyrosine fluorescence parameters of, 64 intensity decays, 578 quantum yield standards, 54 spectral properties, 530–535 Arrays DNA, 732–734 multiplexed microbead:suspension, 726–728 Arsine ligand, 693 9-Arylxanthyl cations, 279 9-AS, 359–360 12-AS, 360, 375 Asparaginase, 548 Associated anisotropy decays, 393, 413–417 Association constant, 282 Association reactions anisotropy, 372–374 helicase-catalyzed DNA unwinding, 373–374 melittin association detected from homotransfer, 374 peptide-calmodulin binding, 372–373 Trp repressor-DNA binding, 373 calmodulin, tryptophan mutants, calcium binding site resolution, 552 energy transfer, 455–456 literature references, 837 orientation of bound peptide, 456–457 Prodan effects on emission spectra, 217–219 quenching, 304–305 accessibility to quenchers, 312–313 substrate binding to ribozymes, 311–312 specific binding reactions, 304–305 Asymptotic standard errors (ASEs), 134–135, 163, 180 ATP, 455 ATPase, single-molecule activity, 770–771 Autocorrelation function, 132–133 Autocorrelation function of fluorescence fluctuations, 800–802, 804–805, 810–811, 813 Avalanche photodiodes (APDs), 114 Avidin, 450, 676 7AW, 564, 565 Azacrown, 79 Azacrown ethers, 644 Azide, 348 Azurin, 536, 546, 547, 583, 588 emission spectra of molecules with one or two tryptophan, 539 fluorescence studies of, 538 site-directed mutagenesis of single-tryptophan, 538–539 B BABAPH, 253–255 BABP, 253–255 Background signal, 366 Bandpass filters, 39 BAPTA, 79, 644, 647–648 BAPTA-based probes, 648–649 Barnase, 556–557 decay-associated spectra, 591–592 β-Barrel, 538 BCECF, 751 Bead sensors, 656 Beam splitter, 29, 777 Beer-Lambert law, deviation from, 58–59 Beer's law, deviation from, 59 Benzene, 230 quenching of indole, 537 Benzo(b)fluoranthene, 632 Benzodiazepines, 23 5,6-Benzoquinoline, 279 Bialkali photocathode, 45–46 Bilins, 84 Bimolecular quenching, 281–282, 294, 304, 547–549 Bimolecular quenching constant, 339 Binding. See Association reactions
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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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578 TIME-RESOLVED PROTEIN FLUORESCE
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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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758 SINGLE-MOLECULE DETECTION Figur
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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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Page 881 and 882: Appendix I Corrected Emission Spect
Page 883 and 884: PRINCIPLES OF FLUORESCENCE SPECTROS
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Page 929: Index A Absorption spectroscopy, 12
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