Imaging fluorescence lifetime

Fluorescence Lifetime Imaging Study on Living Cells with Particular Regard to Electric Field Effects and pH Dependence 607... 

Kuimova MK, Yahioglu G, Levitt JA, Suhling K (2008) Molecular rotor measures viscosity of live cells via fluorescence lifetime imaging. J Am Chem Soc 130(21) 6672-6673... 

Malo GD, Pouwels LJ, Wang M, Weichsel A, Montfort WR, Rizzo MA, Piston DW, Wachter RM (2007) X-ray structure of Cerulean GFP a tryptophan-based chromophore useful for fluorescence lifetime imaging. Biochemistry 46 9865-9873... 

Gadella, T., Jovin, T. and Clegg, R. (1994). Fluorescence lifetime imaging microscopy (FLIM) Spatial resolution of microstructures on the nanosecond time scale. Biophys. Chem. 48, 221-39. 

Holub, O., Seufferheld, M. J., Gohlke, C., Govindjee, G. J., Heiss, G. J. and Clegg, R. M. (2007). Fluorescence lifetime imaging microscopy of Chlamydomonas reinhardtii Non-photochemical quenching mutants and the effect of photosynthetic inhibitors on the slow chlorophyll fluorescence transients. J. Microsc. 226, 90-120. 

Redford, G. and Clegg, R. M. (2005). Real time fluorescence lifetime imaging and FRET using fast-gated image intensifiers. In Methods in... 

While publications on fluorescence lifetime imaging microscopy (FLIM) have been relatively evenly divided between time and frequency domain methods, a majority of the 10 most highly cited papers using FLIM have taken advantage of the frequency domain method [1, 2-9]. Both techniques have confronted similar challenges as they have developed and, as such, common themes may be found in both approaches to FLIM. One of the most important criteria is to retrieve the maximum information out of a FLIM... 

Bastiaens, P. I. H. and Squire, A. (1999). Fluorescence lifetime imaging microscopy Spatial resolution of biochemical processes in the cell. Trends Cell Biol. 9, 48-52. 

Colyer, R. A., Lee, C. and Gratton, E. (2008). A novel fluorescence lifetime imaging system that optimizes photon efficiency. Microsc. Res. Tech. 71, 201-13. 

Fluorescence lifetime imaging Multi-point calibration, minimum resolvable differences, and artifact suppression. Cytometry 43, 248-60. 

Clayton, A. H. A., Hanley, Q. S. and Verveer, P. J. (2004). Graphical representation and multicomponent analysis of single-frequency fluorescence lifetime imaging microscopy data. J. Microsc. 213, 1-5. 

Anonymous. (2003). LIFA system for fluorescence lifetime imaging microscopy (FLIM). J. Fluoresc. 13, 365-7. 

Hanley, Q. S., Arndt-Jovin, D. J. and Jovin, T. M. (2002). Spectrally resolved fluorescence lifetime imaging microscopy. Appl. Spectrosc. 56,155-66. 

Hanley, Q. S. and Ramkumar, V. (2005). An internal standardization procedure for spectrally resolved fluorescence lifetime imaging. Appl. Spectrosc. 59, 261-6. 

The lifetime of the excited state of fluorophores may be altered by physical and biochemical properties of its environment. Fluorescence lifetime imaging microscopy (FLIM) is thus a powerful analytical tool for the quantitative mapping of fluorescent molecules that reports, for instance, on local ion concentration, pH, and viscosity, the fluorescence lifetime of a donor fluorophore, Forster resonance energy transfer can be also imaged by FLIM. This provides a robust method for mapping protein-protein interactions and for probing the complexity of molecular interaction networks. 

At the end of the 1980s and early 1990s, first experiments were carried out to combine fluorescence lifetime measurements with imaging using both time domain [1-4] and frequency domain [5-7] based approaches. This chapter will deal exclusively with time domain based fluorescence lifetime imaging methods. For the frequency domain based methods, refer Chapter 2. 

Lifetime imaging can be implemented both in wide field and in scanning microscopes such as confocal microscopes and two-photon excitation microscopes. The most common implementations in time-domain fluorescence lifetime imaging microscopy (FLIM) are based on TCSPC [8, 9] and time-gating (TG) [2, 10],... 

Conventional TCSPC equipment has been successfully employed in LSM for fluorescence spectroscopy on discrete microscopic volumes [18, 19] and for fluorescence lifetime imaging at a low acquisition speed [1], The use of conventional TCSPC equipment for imaging results in very long acquisition times, several to many minutes per (time-resolved) image. Importantly, operating the TCSPC detection system at too high detection rates, above 5% of the excitation frequency, results in distortion of the recorded decay curve [20],... 

Also global fitting techniques, where the space invariance (or any other invariance property) of one or more fitting parameters is exploited, have been successfully used to analyze fluorescence lifetime images [45, 46]. When applicable, global analysis techniques provide more homogeneous SNRs and reduce the number of fitted parameters. 

Wang, X. F., Uchida, T., Coleman, D. M. and Minami, S. (1991). A 2-dimensional fluorescence lifetime imaging-system using a gated image intensifier. Appl. Spectrosc. 45, 360-6. 

Becker, W., Bergmann, A., Hink, M. A., Konig, K., Benndorf, K. and Biskup, C. (2004b). Fluorescence lifetime imaging by time-correlated single-photon counting. Microsc. Res. Tech. 63, 58-66. 

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