Principles of Fluorescence Spectroscopy

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880 APPENDIX I P CORRECTED EMISSION SPECTRA Table I.8. Corrected Emission Spectra in Relative Quanta per Wavelength Interval f Quinine sulfate b 3-APT c N,N-DMANB d 4,4'-DMANS e λ (nm) a I(λ) λ (nm) I(λ) λ (nm) I(λ) λ (nm) I(λ) 384.6 1.4 434.8 1.4 444.4 2.2 555.6 2.6 388.3 3.5 439.6 2.0 449.4 2.9 563.4 3.4 392.2 5.5 444.4 4.0 454.5 4.2 571.4 4.1 396.0 8.7 449.4 7.7 459.8 8.3 579.7 6.4 400.0 13.8 454.5 13.9 465.1 14.2 588.2 9.4 404.0 19.4 459.8 21.5 470.6 21.1 597.0 13.6 408.2 26.6 465.1 33.7 476.2 30.2 606.1 19.1 412.4 36.6 470.6 46.4 481.9 40.8 615.4 24.9 416.7 45.5 476.2 60.8 487.8 50.9 625.0 33.2 421.1 54.7 481.9 74.0 493.8 61.0 634.9 42.8 425.5 64.6 487.8 84.8 500.0 71.2 645.2 53.2 430.1 74.6 493.8 93.4 506.3 81.4 655.7 64.0 434.8 82.5 500.0 98.4 512.8 88.7 666.7 74.7 439.6 90.0 506.3 100.0 519.5 94.1 678.0 84.5 444.4 95.0 512.8 99.0 526.3 98.5 689.7 91.9 449.4 98.6 519.5 95.0 533.3 100.0 701.8 96.4 454.5 100.0 526.3 89.2 540.5 99.3 714.3 99.4 459.8 99.2 533.3 82.3 547.9 96.7 727.3 100.0 465.1 97.5 540.5 73.5 555.6 92.2 740.7 98.4 470.6 93.8 547.9 63.3 563.4 87.3 754.7 93.3 476.2 88.3 555.6 54.8 571.4 81.8 769.2 86.7 481.9 81.7 563.4 46.3 579.7 75.5 784.3 78.1 487.8 74.9 571.4 39.9 588.2 69.6 800.0 67.9 493.8 67.9 579.7 34.1 597.0 63.8 816.3 57.1 500.0 60.3 588.2 29.0 606.1 58.0 833.3 46.6 506.3 53.4 597.0 24.5 615.4 52.4 851.1 37.6 512.8 46.9 606.1 20.9 625.0 45.9 869.6 29.6 519.5 41.0 615.4 17.5 634.9 40.2 888.9 22.2 526.3 35.0 625.0 14.7 645.2 35.0 909.1 16.0 533.3 30.0 634.9 12.3 655.7 30.5 930.2 11.5 540.5 24.9 645.2 10.0 666.7 26.6 952.4 7.4 547.9 20.0 655.7 7.9 678.0 22.5 555.6 16.4 666.7 5.9 689.7 19.0 563.4 13.6 678.0 4.2 701.8 16.3 571.4 11.6 689.7 2.7 714.3 13.4 579.7 10.0 701.8 1.6 727.3 11.0 588.2 8.5 714.3 0.8 740.7 9.0 597.0 6.8 754.7 6.9 606.1 5.5 769.2 5.4 615.4 4.2 784.3 4.0 625.0 3.2 800.0 2.7 634.9 2.4 816.3 1.8 645.2 1.5 833.3 0.8 655.7 0.7 666.7 0.0 a Calculated from Table I.7. b Quinine sulfate (10 –3 M) in 0.1 N H2 SO 4 , 20°C. c 3-aminophthalimide (5 x 10 –4 M) in 0.1 N H2 SO 4 , 20°C. d N,N'-dimethylamino-m-nitrobenzene (10 –4 M) in 30% benzene, 70% m-heptane, 20°C e 4-dimethylamino-4'-nitrostilbene in o-dichlorobenzene, 20°C. f Calculated from Table I.7 using I(λ) = λ –2 I(ν), followed by normalization of the peak intensity to 100.
PRINCIPLES OF FLUORESCENCE SPECTROSCOPY 881 Table I.9. Corrected Emission Spectrum of 2- Aminopyridine a ν (cm –1 ) I(ν) λ (nm) I(λ) b 20,800 0.010 480.8 0.006 21,500 0.038 465.1 0.024 22,200 0.073 450.5 0.049 23,000 0.133 434.8 0.095 23,800 0.264 420.2 0.202 24,700 0.450 404.9 0.371 25,600 0.745 390.6 0.660 26,600 0.960 375.9 0.918 27,200 1.00 367.7 1.00 27,800 0.939 359.7 0.981 28,900 0.587 346.0 0.663 30,150 0.121 331.7 0.149 31,000 0.033 322.6 0.049 a 10 –5 M in 0.1 N2 SO 4 . From [10] and [11]. b Calculated using I(λ) = I(ν)λ –2 followed by normalization of the maximum to unity. (Table I.9 and Figure I.10). Corrected emission spectra have been reported 10 for phenol and the aromatic amino acids, phenylalanine, tyrosine and tryptophan (Figure I.11). 6. ADDITIONAL CORRECTED EMISSION SPECTRA Corrected spectra as I(ν) versus ν can be found in the compendium by Berlman. 11 Included in that volume are a number of UV-emitting species including indole and phenol, which can be used to obtain corrected emission spectra of proteins. For convenience Berlman's spectra for phenol and Figure I.10. Corrected emission spectra of 2-aminopyridine in relative quanta per wavenumber interval I(ν) and per wavelength interval I(λ). See Table I.8 for additional details. From [8] and [9]. Figure I.11. Corrected emission spectrum (I(λ)) for phenylalanine (!), phenol (△), tyrosine (), and tryptophan ("). The areas underneath the curves are proportional to the quantum yields. Data from [10]. Figure I.12. Corrected emission spectra of phenol in methanol and indole in ethanol. From [11]. indole are provided as Figure I.12. Cresyl violet in methanol has been proposed as a quantum yield and emission spectral standard for red wavelengths. 12 In methanol at 22°C the quantum yield of cresyl violet is reported to be 0.54, with an emission maximum near 614 nm. The use of quinine as a standard has occasionally been questioned. 13–16 Additional discussion about corrected emission spectra can be found in [17–20]. REFERENCES 1. Gardecki JA, Maroncelli M. 1998. Set of secondary emission standards for calibration of the spectral responsibility in emission spectroscopy. Appl Spectrosc 52:1179–1189.
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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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16 The biochemical applications of
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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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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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794 SINGLE-MOLECULE DETECTION Polar
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Principles of Fluorescence Spectroscopy

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