Principles of Fluorescence Spectroscopy

PRINCIPLES OF FLUORESCENCE SPECTROSCOPY 931
DPPG vesicles, 408, 691
DsRed, 82
DTAC micelles, 286
Dual-color cross fluorescence correlation spectroscopy, 823–828
Dye lasers, 110–112
Dyes. See also specific dyes
energy-transfer, 709–710
red and near-IR, 74–75
solvent relaxation, 253
Dynamic quenching. See Collisional quenching; Quenching
Dynamics of relaxation. See Relaxation dynamics
Dynode chains, 44–45, 46–47
Dynorphin, 590
E
EcoRI, 826
EDANS, 721
Efficiency, quenching, 281, 333–334, 542
EGFP (excited green fluorescent protein)
fluorescence correlation spectroscopy, 816–817
EIA 5-(iodoacetamido)eosin
Förster distances, 468
Einstein equation, 12
Electrical bias, 166
Electron-exchange quenching, 335
Electronics
frequency-domain fluorometers, 164–165
photomultiplier tube, 44–45
time-correlated single-photon counting, 114–117
Electronic state resolution from polarization spectra, 360–361
Electronic states, 1, 5
of metal–ligand complexes, 684–685
Electron scavengers, quenching by, 278
Electron transfer
photoinduced (PET), 335–336, 627
quenching in flavoprotein, 315–317
Electrooptic modulators, 165–166
ELISA, 659
Ellipsoids, 418–419
anisotropy decay, 419–420
of revolution, 420–425
rotational diffusion of, 425–426
Elongation factor (Ta-GDP), 548
Emission center of gravity, 246–247
Emission filter, 41
Emission spectra, 3–4, 7–8
decay-associated, 591–592, 596–598
instrumentation, 27–31
distortions in, 30–31
for high throughput, 29–30
ideal spectrofluorometer, 30
molecular information from fluorescence, 17–18
phase-sensitive, 194–197
solvent effects, 219–221
through optical filters, 43–44
tryptophan decay-associated, 581
Emission spectra, corrected
instrumentation, 52–54
comparison with known emission spectra, 52–53
conversion between wavelength and wavenumber, 53–54
correction factors obtained with standard lamp, 53
quantum counter and scatterer use, 53
Emission spectra, quenching-resolved, 301–304
fluorophore mixtures, 301–302
Tet repressor, 302–304
Emission wavelength, and anisotropy, 437–438
Encounter complexes, 333–334
Endonuclease, 290–291
Energy gap law, 687–688
Energy transfer, 443–468
biochemical applications, 453–458
intracellular protein folding, 454–455
orientation of protein-bound peptide, 456–457
protein binding to semiconductor nanoparticles, 457–458
protein folding measured by RET, 453–454
RET and association reactions, 455–456
data analysis, 485–487
diffusion-enhanced, 467
distance distributions, 477–479
distance measurements
melittin, α helical, 451–452
distance measurements using RET, 451–453
incomplete labeling effects, 452
orientation factor effects on possible range of distances,
448–449, 452–453
DNA hybridization, one donor- and acceptor-labeled probe, 717–718
effect of colloids, 847
energy-transfer efficiency from enhanced fluorescence, 461–462
Förster distances, 443–444, 445–446, 468
GFP sensors, 654–655
homotransfer, 450
incomplete labeling effects, 452
lanthanides, 680–681
long-wavelength long-lifetime fluorophores, 696
in membranes, 462–465
lipid distributions around Gramicidin, 463–465
membrane fusion and lipid exchange, 465
nucleic acids, 459–461
imaging of intracellular RNA, 460–461
orientation factor, 465
protein kinase C activation, fluorescence-lifetime imaging
microscopy, 746–747
proteins, 539–545
anisotropy decreases, detection of ET by, 531–532
interferon-γ, tyrosine-to-tryptophan energy transfer in, 540–541
phenylalanine-to-tyrosine, 543–545
RET efficiency quantitation, 541–543
proteins as sensors, 652–654
quantum dots, 678
radiationless, anisotropy measurement, 365
red-edge excitation shifts, 259
representative R 0 values, 467–468
RET sensors, 458–459
imaging of intracellular protein phosphorylation, 459
intracellular RET indicator for estrogens, 458
Rac activation in cells, 459
sensor mechanisms, 633–637
glucose, 634–635
ion, 635–636
pH and CO 2 , 633–634
theory for, 636–637
silver particles effect, 854–855
single-molecule detection, 773–775
literature references, 794
in solution, 466–467