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Introduction Figure 1.16: On excitation of a fluorophore by a modulated light source the emission is shifted in phase by φ and demodulated according to the relationship for m shown. (a) (b) Figure 1.17: (a) Emission of a 5 ns decay time fluorophore in response to modulated excitation at 2.5, 25, and 250 MHz. (b) Comparison of a 10 ns decay with modulation frequencies of 2.5 to 250 MHz. Reproduced from [36]. 30
Introduction case, the molecules excited at the peak of the intensity continue to emit during the entire excitation cycle. The resultant averaging of the decay across the cycle gives an increasing phase shift and a greater demodulation of the emission signal. The influence of different lifetimes on the phase and demodulation responses is shown in the simulated FD curves in Figure 1.18. The phase and demodulation curves cross at low modulation frequencies for short lifetime fluorophores and at higher modulation frequencies for longer lifetime fluorophores. The cross-over point gives an indication of the optimal modulation frequency to be used to excite the sample as this is where the maximum rate of change in phase and demodulation occurs. Figure 1.18: Simulated FD data (square: modulation and circle: phase) for single exponential decays of lifetime 2.5 (yellow points) and 10 ns (blue points). The phase shift increases and the modulation decreases with increasing modulation frequency. The theory of the FD method are described in many publications [36, 53, 54] and the complete derivation is reproduced here. Referring back to Figure 1.16, if the excitation is sinusoidally modulated light, then: I0(t) = a + b sin ωt (1.20) 31
Page 1 and 2: Time-Resolved Fluorescence Spectros
Page 3 and 4: Abstract behaviour allowing calcula
Page 5 and 6: Contents List of Publications and P
Page 7 and 8: Contents 2.3 Temperature based Life
Page 9 and 10: List of Publications and Presentati
Page 11 and 12: Abbreviations Abbrev Definition API
Page 13 and 14: Chapter 1 Introduction For many yea
Page 15 and 16: Introduction temperature increases
Page 17 and 18: A seal or cap-rock that prevents th
Page 19 and 20: Introduction Figure 1.2: Crude oil
Page 21 and 22: Introduction eroatoms (O, S, N) and
Page 23 and 24: Introduction toluene, dichlorometha
Page 25 and 26: 1.2 Fluorescence Spectroscopy 1.2.1
Page 27 and 28: Introduction S1 state for a certain
Page 29 and 30: Introduction The peaks in the absor
Page 31 and 32: Introduction Figure 1.9: Jablonski
Page 33 and 34: Introduction Figure 1.10: Static qu
Page 35 and 36: Introduction the ratio of τ 0/τ i
Page 37 and 38: 1.2.5 Measuring fluorescence lifeti
Page 39 and 40: Introduction Figure 1.14: Time-reso
Page 41: Introduction The autocorrelation fu
Page 45 and 46: From Equation 1.26, Introduction ω
Page 47 and 48: m = 1 P 2 + Q 2 Introduction (1.36
Page 49 and 50: Introduction based average lifetime
Page 51 and 52: Introduction Figure 1.21: General p
Page 53 and 54: Introduction TAC output pulse is gi
Page 55 and 56: Introduction at each frequency acco
Page 57 and 58: Introduction diagram showing the op
Page 59 and 60: Introduction Figure 1.26: Schematic
Page 61 and 62: Introduction provides enhanced cont
Page 63 and 64: Introduction PAH’s. It has been s
Page 65 and 66: to heavy oils (low API gravity). In
Page 67 and 68: Introduction The balance between en
Page 69 and 70: Introduction Figure 1.34: Fluoresce
Page 71 and 72: Introduction used [126, 129, 132] a
Page 73 and 74: Introduction Figure 1.36: Plot of a
Page 75 and 76: fractional contributions over time
Page 77 and 78: Introduction concentration and Ksv
Page 83 and 84: 1.5 Crude oil photophysics Introduc
Page 85 and 86: Introduction stages of thermal matu
Page 87 and 88: Introduction yellow to red fluoresc
Page 89 and 90: 1.7 Thesis goal Introduction This i
Page 91 and 92: Materials and Methods concentration
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Table 2.2: Fractionation Data for B
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Materials and Methods 2.2 Frequency
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Materials and Methods was within ±
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Materials and Methods The dichroic
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Figure 2.5: Transmission characteri
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Materials and Methods Figure 2.7: S
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Materials and Methods The variation
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Materials and Methods Figure 2.9: L
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Materials and Methods Figure 2.10:
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Materials and Methods Comparison ca
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Materials and Methods with ρ param
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Materials and Methods 2.3 Temperatu
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Materials and Methods IRF of the sy
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Materials and Methods 2.4 Temperatu
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Materials and Methods a series of w
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Materials and Methods 2. Using the
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Lifetime study on crude oils by the
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Table 3.3: Average lifetime and sin
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Table 3.5: Average lifetime and sin
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3.5 Single Lifetime distributions L
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Energy Transfer and Quenching proce
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Influence of low temperature on flu
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Conclusions and Future Work model (
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Appendix A Reference Lifetime Stand
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Appendix Figure A.2: FD response fo
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A.3 SPA N-(3-sulfopropyl) acridiniu
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A.4 Rhodamine B Appendix Rhodamine
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A.5 Acridone Appendix The highly ph
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Appendix Figure A.11: FD response f
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Table B.1: Frequency Domain average
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Appendix Table B.3: Emission λmax
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Table B.5: Frequency Domain lifetim
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Appendix for the determination of t
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Appendix Figure C.3: Multifrequency
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Appendix Figure C.5: FLIM intensity
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Appendix Table C.1: Fluorescence li
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Appendix D Published Work 281
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998 J Fluoresc (2008) 18:997-1006 s
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1000 J Fluoresc (2008) 18:997-1006
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1002 J Fluoresc (2008) 18:997-1006
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1004 J Fluoresc (2008) 18:997-1006
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1006 J Fluoresc (2008) 18:997-1006
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Bibliography [10] W. G. Dow (1977),
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Bibliography [31] M. A. Ali and W.
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Bibliography [53] R. D. Spencer and
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Bibliography [75] K. Konig (2000),
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Bibliography [93] C. Pasquini and A
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Bibliography [113] C. Ralston, X. W
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Bibliography [132] A. Ryder (2004),
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Bibliography variable-angle synchro
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Bibliography [171] R. K. McLimans (
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Bibliography using fluorescence spe
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Bibliography [210] H. Mishra, S. Pa
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Bibliography [229] N. Boens, W. W.
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