Hyper-spectral modulation fluorescent imaging using double acousto-optical tunable filter based on TeO2-crystals

Abstract

We have proposed a method for hyper-spectral fluorescent imaging based on acousto-optical filtering. The object of interest was pumped using ultraviolet radiation of mercury lamp equipped with monochromatic excitation filter with the window of transparency centered at 365 nm. Double TeO2-based acousto-optical filter, tunable in range from 430 to 780 nm and having 2 nm bandwidth of spectral transparency, was used in order to detect quasimonochromatic images of object fluorescence. Modulating of ultraviolet pump intensity was used in order to reduce an impact of non-fluorescent background on the sample fluorescent imaging. The technique for signal-to-noise ratio improvement, based on fluorescence intensity estimation via digital processing of modulated video sequence of fluorescent object, was introduced. We have implemented the proposed technique for the test sample studying and we have discussed its possible applications.