Principles of Fluorescence Spectroscopy

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