Fluorescence Lifetime Imaging Microscopy FLIM

What type of chemical imaging will be possible using FLIM technology Based on our current understanding of FLIM, and factors that affect fluorescence lifetimes, 

Chromosome imaging Acridine lifetimes depend on DNA base composition 

Microviscosity imaging Identify viscosity-lifetime probes 

Proximity imaging by energy transfer Protein-protein binding Protein-membrane association 

Abbreviations NADH, nicotinamide-adenine dinucleotide phosphate TNS, 2-(p-toluidino)naphthalene-6-sulfonic acid. 

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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. 

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... 

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

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],... 

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

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). 

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. 

Figure 1.8. Fluorescence lifetime imaging microscopy (FLIM).

R.R. Duncan, A. Bergmann, M.A. Cousin, D.K. Apps, M.J. Shipston, Multidimensional time-correlated single-photon counting (TCSPC) fluorescence lifetime imaging microscopy (FLIM) to deetct FRET in cells, J. Microsc. 215, 1-12 (2004)... 

R.V. Krishnan, H. Saitoh, H. Terada, V.E. Centonze, B. Herman, Development of a multiphoton fluorescence lifetime imaging microscopy (FLIM) system using a streak camera. Rev. Sci. Instrum. 74, 2714-2721 (2003)... 

P. Tinnefeld, V. Buschmann, D-P. Herten, K.T. Han, M. Sauer, Confocal fluorescence lifetime imaging microscopy (FLIM) at the single molecule level, Single Mol. 1, 215-223 (2000)... 

Fluorescence lifetime imaging microscopy (FLIM) measures a chromophore s fluorescence lifetime, allowing spatial resolution of biochemical processes. The fluorescence lifetime of a donor dye decreases under FRET conditions, independent of fluorophore concentration or of excitation intensity (35). 

Fluorescence Lifetime Imaging Microscopy (FLIM) Measurement... 

While phase-soisitive detection of fluorescence is somewhat outdated for the resolution of multicomponent mixtures, this mediod has found use in fluorescence lifetime imaging microscopy (FLIM). The concept of FLIM is illusirated in Figure 22.16. Suppose that die sample contains two regions in which die fluorophore displays different lifetimes, T and tj. The intensity from both regions may be the same, so dial die difference in probe environment is not visible in die steady-state image. If the lifetimes... 

Fluorescence lifetime imaging microscopy (FLIM)-based guantitative fluorescence resonance energy transfer (FRET). Direct detection of biomolecular interactions is possible with FRET measurements, where a donor fluorophore transfers the energy to an acceptor fluorophore in case they are close in space. The combination of this technique together with FLIM, based on the decrease in donor fluorophore lifetime that is induced by FRET, has recently enabled the quantitative assessment of the protein-interacting fractions [12]. 

In order to determine the efficacy of palmostatin B to inhibit APTl in cells, fluorescence lifetime imaging microscopy (FLIM) was performed with... 

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