Fluorescence lifetime imaging microscopy, FLIM, is a technique for producing an image based on the differences in the exponential decay rate (lifetime) of the fluorescent molecule.
FLIM is an easy and fast application to image FRET as the lifetime of the donor decreases upon FRET. CFP-YFP and GFP-DsRed pairs have been successfully used.
Our Lambert Instruments LIFA is a fast frequency domain FLIM system, which is attached to an inverted Carl Zeiss AxioImager microscope body. With this system we can image only fixed samples in widefield mode. The light source is multi-LED with excitation lines 406, 469 and 533nm. With the LIFA system we can image lifetimes with a resolution < 100 ps in a range of 0.1 ns – 1 ms.
The system is capable of high-content imaging.
For basic information, please read ‘Fluorescence lifetime imaging microscopy: spatial resolution of biochemical processes in the cell’ by Bastiaens and Squire. (Trends Cell Biol. 1999 Feb;9(2):48-52.)
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