analyze bi-exponential decays. One major drawback of this method is that it cannot take into account the instrument response effect and for this reason the early part of the measured decay curves should be ignored in the analyses. This means that part of the signal is discarded and the accuracy for estimating short lifetimes goes down. One of the interesting features of the convolution theorem is that the integral of the convolution is the product of the factors that make up the integral. There are a few techniques which work in transformed space that exploit this property to recover the pure decay curve from the measured curve. Laplace and Fourier transformation along with Laguerre gauss expansion have been used to estimate the lifetime in transformed space. These approaches are faster than the deconvolution based methods but they suffer from truncation and sampling problems. Moreover, application of methods like Laguerre gauss expansion is mathematically complicated. In Fourier methods the lifetime of a single exponential decay curve is given by: Where: and n is the harmonic number and T is the total time range of detection. Applications FLIM has primarily been used in biology as a method to detect photosensitizers in cells and tumors as well as FRET in instances where ratiometric imaging is difficult. The technique was developed in the late 1980s and early 1990s (Gating method: Bugiel et al. 1989. König 1989, Phase modulation: Lakowicz at al. 1992,) before being more widely applied in the late 1990s. In cell culture, it has been used to study EGF receptor signaling and trafficking. Time domain FLIM (tdFLIM) has also been used to show the interaction of both types of nuclear intermediate filament proteins lamins A and B1 in distinct homopolymers at the nuclear envelope, which further interact with each other in higher order structures. FLIM imaging is particularly useful in neurons, where light scattering by brain tissue is problematic for ratiometric imaging. In neurons, FLIM imaging using pulsed illumination has been used to study Ras, CaMKII, Rac, and Ran family proteins. FLIM has been used in clinical multiphoton tomography to detect intradermal cancer cells as well as pharmaceutical and cosmetic compounds. More recently FLIM has also been used to detect flavanols in plant cells. Autofluorescent coenzymes NAD(P)H and FAD Multi-photon FLIM is increasingly used to detect auto-fluorescence from coenzymes as markers for changes in mammalian metabolism. FRET imaging Since the fluorescence lifetime