Abstract
Time-tagged TCSPC (time-correlated single photon counting) is a special acquisition mode of TCSPC with which one determines not only the excitation-emission delay time of detected photons but also their arrival times measured from the start of the experiment. Time-tagged TCSPC enables us to examine slow fluctuation of fluorescence lifetimes, which is particularly important in the study of heterogeneous or fluctuating systems at the single-molecule level. In this chapter, we describe recent development of new methods using time-tagged TCSPC, aiming at showing their high potential in studying dynamics of complex systems. We depict two closely related methods based on fluorescence correlation spectroscopy (FCS), i.e., lifetime-weighted FCS and two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS). These methods enable us to quantify fluorescence lifetime fluctuations on the microsecond timescale. Showing examples including the study of a biological macromolecule, we demonstrate the usefulness of these two methods in real applications. In addition, we present another application of time-tagged TCSPC, which analyzes photon interval time for characterizing timing instability of photon detectors.
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References
Becker W (2005) Advanced time-correlated single photon counting techniques. Springer series in chemical physics, vol 81. Springer, Heidelberg
Eggeling C, Fries JR, Brand L, Günther R, Seidel CAM (1998) Monitoring conformational dynamics of a single molecule by selective fluorescence spectroscopy. Proc Natl Acad Sci U S A 95:1556–1561
Palo K, Brand L, Eggeling C, Jaeger S, Kask P, Gall K (2002) Fluorescence intensity and lifetime distribution analysis: toward higher accuracy in fluorescence fluctuation spectroscopy. Biophys J 83:605–618
Yang H, Xie XS (2002) Probing single-molecule dynamics photon by photon. J Chem Phys 117:10965–10979
Boehmer M, Wahl M, Rahn H-J, Erdmann R, Enderlein J (2002) Time-resolved fluorescence correlation spectroscopy. Chem Phys Lett 353:439–445
Gregor I, Enderlein J (2007) Time-resolved methods in biophysics. 3. Fluorescence lifetime correlation spectroscopy. Photochem Photobiol Sci 6:13–18
Kapusta P, Wahl M, Benda A, Hof M, Enderlein J (2007) Fluorescence lifetime correlation spectroscopy. J Fluoresc 17:43–48
Kapusta P, Macháň R, Benda A, Hof M (2012) Fluorescence lifetime correlation spectroscopy (FLCS): concepts, applications and outlook. Int J Mol Sci 13:12890–12910
Ishii K, Tahara T (2010) Resolving inhomogeneity using lifetime-weighted fluorescence correlation spectroscopy. J Phys Chem B 114:12383–12391
Ishii K, Tahara T (2012) Extracting decay curves of the correlated fluorescence photons measured in fluorescence correlation spectroscopy. Chem Phys Lett 519–520:130–133
Ishii K, Tahara T (2013) Two-dimensional fluorescence lifetime correlation spectroscopy. 1. Principle. J Phys Chem B 117:11414–11422
Ishii K, Tahara T (2013) Two-dimensional fluorescence lifetime correlation spectroscopy. 2. Application. J Phys Chem B 117:11423–11432
Otosu T, Ishii K, Tahara T (2013) Note: simple calibration of the counting-rate dependence of the timing shift of single photon avalanche diodes by photon interval analysis. Rev Sci Instrum 84:036105
Lakowicz JR (2006) Principles of fluorescence spectroscopy, 3rd edn. Springer, New York
Yang H, Xie XS (2002) Statistical approaches for probing single-molecule dynamics photon-by-photon. Chem Phys 284:423–437
Burstyn HC, Sengers JV (1983) Time dependence of critical concentration fluctuations in a binary liquid. Phys Rev A 27:1071–1085
Livesey AK, Brochon JC (1987) Analyzing the distribution of decay constants in pulse-fluorimetry using the maximum entropy method. Biophys J 52:693–706
Brochon JC (1994) Maximum entropy method of data analysis in time-resolved spectroscopy. Methods Enzymol 240:262–311
Hanbury Brown R, Twiss RQ (1956) Correlation between photons in two coherent beams of light. Nature 177:27–29
Berglund AJ, Doherty AC, Mabuchi H (2002) Photon statistics and dynamics of fluorescence resonance energy transfer. Phys Rev Lett 89:068101
Nettels D, Gopich IV, Hoffmann A, Schuler B (2007) Ultrafast dynamics of protein collapse from single-molecule photon statistics. Proc Natl Acad Sci U S A 104:2655–2660
Rech I, Labanca I, Ghioni M, Cova S (2006) Modified single photon counting modules for optimal timing performance. Rev Sci Instrum 77:033104
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Ishii, K., Otosu, T., Tahara, T. (2014). Lifetime-Weighted FCS and 2D FLCS: Advanced Application of Time-Tagged TCSPC. In: Kapusta, P., Wahl, M., Erdmann, R. (eds) Advanced Photon Counting. Springer Series on Fluorescence, vol 15. Springer, Cham. https://doi.org/10.1007/4243_2014_65
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DOI: https://doi.org/10.1007/4243_2014_65
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