Gryczynski, Zygmunt Karol2020-12-262020-12-2620202020https://repository.tcu.edu/handle/116099117/42825Fluorescence technology has enabled many practical applications in chemistry, biology, and medicine. However, the fluorescence intensity dependence on fluorophore absorption/concentration can develop non-linearity because of the Primary Inner-Filter Effect (IFE). The primary IFE is a consequence of the progressive excitation light attenuation due to sample absorption. For low sample concentrations/absorptions, this effect is minimal, yet still detectable (even for absorbances < 0.05). The Inner-Filter Effect has been recognized for many decades, yet there is no method that adequately corrects the loss in linearity. Hence, I am proposing an original strategy based on the calculation of a spectrofluorometer’s “geometrical sensitivity factor”, which can be found for any instrument. Using a thin layer sample (= 1 mm) that is mounted on a movable holder, we can determine the sensitivity factor that will allow us to correct the emission/excitation spectra and restore the linear dependence.Format: OnlineenPhysicsBiophysicsFluorescenceInner-filter effectText