Fluorescence lifetime imaging microscopy

The usehilness of FLIM is a result of the difficulties of quantifying fluorescence intensities in fluorescence microscopy. The steady-state fluorescence images can be difficult to interpret and quantify because there is no prac- 

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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. 

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. 

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. 

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. 

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],... 

Pelet, S., Previte, M. J., Laiho, L. H. and So, P. T. (2004). A fast global fitting algorithm for fluorescence lifetime imaging microscopy based on image segmentation. Biophys. J. 87, 2807-17. 

Wang, X. F., Periasamy, A. and Herman, B. (1992). Fluorescence lifetime imaging microscopy (FLIM) Instrumentation and applications. Crit. Rev. Anal. Chem. 23, 369-95. 

Total internal reflection fluorescence lifetime imaging microscopy... 

Van Munster, E. B. and Gadella, T. W. J., Jr. (2004). phiFLIM A new method to avoid aliasing in frequency-domain fluorescence lifetime imaging microscopy. J. Microsc. 213, 29-38. 

Fluorescence lifetime imaging microscopy (FLIM) that measures the donor excited state lifetime in the presence and absence of an acceptor [23, 25, 28, 47, 52, 53, 57, 58] (see Chapters 2-4). 

Elangovan, M., Day, R. N. and Periasamy, A. (2002). Nanosecond fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy to localize the protein interactions in a single living cell. J. Microsc. 205, 3-14. 

Measuring FRET by fluorescence lifetime imaging microscopy (FRET-FLIM) offers the ability to see beyond the resolution of the optical system ( 10-100 times that of modern far field microscopes [5]). FRET efficiency can be used as a proxy for molecular distance, thereby allowing the easy detection and somewhat more challenging quantification of molecular interactions. Although many types of assay exist, FRET-FLIM is a highly suitable technique that is capable of in situ measurements of molecular interactions and conformation in living and fixed cells. 

Sanders, R., Draaijer, A., Gerritsen, H. C., Houpt, P. M. and Levine, Y. K. (1995). Quantitative pH imaging in cells using confocal fluorescence lifetime imaging microscopy. Anal. Biochem. 227, 302-8. 

Tramier, M., Zahid, M., Mevel, J. C., Masse, M. J. and Coppey-Moisan, M. (2006). Sensitivity of CFP/YFP and GFP/mCherry pairs to donor photobleaching on FRET determination by fluorescence lifetime imaging microscopy in living cells. Microsc. Res. Tech. 69, 933-9. 

Peltan, I. D., Thomas, A. V., Mikhailenko, I., Strickland, D. K., Hyman, B. T. and von Arnim, C. A. (2006). Fluorescence lifetime imaging microscopy (FLIM) detects stimulus-dependent phosphorylation of the low density lipoprotein receptor-related protein (LRP) in primary neurons. Biochem. Biophys. Res. Commun. 349, 24-30. 

Calleja, V., Ameer-Beg, S. M., Yojnovic, B., Woscholski, R., Downward, J. and Larijani, B. (2003). Monitoring conformational changes of proteins in cells by fluorescence lifetime imaging microscopy. Biochem. J. 372, 33 40. 

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