Fluorescence lifetime determination of photo-bleaching correlation

FLIM is relatively stable about photo-bleaching effect. In case of dye has multi-exponential decay property, fluorescence lifetime can be affected by photo-bleaching. Such lifetime variation can cause critical error in many FLIM applications. If lifetime is changed by photo-bleaching, we can´t measure accurately donor lifetime with acceptor. It is difficult to separate lifetime variation by photo-bleaching and FRET. For this reason we carry out the experiment to know influence of such fluorescence intensity loss on fluorescence lifetime.