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Introduction Figure 1.13: Simulated time domain traces for a fluorophore that exhibits a 1, 5, and 10 ns excited-state lifetime. be fitted to a sum of exponentials: I(t) = n αie −t/τi (1.14) i=0 where αi is the pre-exponential factor denoting the contribution to the total time- resolved decay of the component with lifetime τi. Figure 1.14 shows situation where there are two fluorophores with different lifetimes, one short and one long lifetime component. The fraction of the steady state intensity due to each component is given by: fi = αiτi j αjτj (1.15) For TD experiments, the intensity of the decay (I(t)) is convolved with the Instru- ment Response Function (IRF). The IRF represents the response of the measure- ment system to the excitation pulse and can be generated by recording the output from a sample that scatters the excitation light or a fluorophore of very short lifetime. The convolution of the (IRF(t)) and the sample decay results in an observed 26
Introduction Figure 1.14: Time-resolved decay of fluorescence intensity of a mixture of two species, the lifetimes of which are represented by τ 1 and τ 2. (N(t)) decay profile of the form: N(t) = n 0 IRF(t)I(t − t ′ )dt ′ (1.16) where t ′ defines the variable time delays and the infinitesimally small time widths dt ′ of which I(t) is composed. The goal is then to extract I(t), and, in turn, the kinetic parameters from the experimental measurables (N(t) and IRF(t)). Numerous methods (such as Fourier and Laplace transforms) exist to extract I(t) from N(t) and IRF(t) [44]. The most common approach is to use a least-squares method in concert with an iterative reconvolution scheme [45, 46]. In this approach, a test model is selected (Equation 1.14), and then the convolution integral (Equation 1.16) is calculated based on an initial set of αi and τi values and the measured IRF. The calculated response (C(t)) is compared with the observed data (N(t)) and the αi and τi terms adjusted until a best fit is obtained. The quality of the fit is judged by the chi-squared (χ 2 ) (goodness of fit) test: χ 2 np = Wj [N(t) − C(t)] 2 j=1 27 (1.17)
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: 1.2.5 Measuring fluorescence lifeti
Page 41 and 42: Introduction The autocorrelation fu
Page 43 and 44: Introduction case, the molecules ex
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
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1.7 Thesis goal Introduction This i
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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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