Fluorescence Lifetime Imaging Microscopy (FLIM)

Fluorescence lifetime-resolved imaging microscopy (FLIM) refers to the technique of image formation in which the fluorescence lifetimes of emitting fluorophores are resolved for every image location in a specimen. By utilizing differences in excited-state lifetimes of the fluorophores as the contrast mechanism for image formation, many of the limitations of conventional fluorescence microscopy methods, based on spectrally resolved fluorescence intensity measurements, are avoided. The information represented in lifetime-resolved images may be the actual (or apparent) lifetimes, or another spectroscopic parameter that is able to be determined only by virtue of the fluorescence signal being lifetime-resolved.

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