Fluorescence Reduction by a Triplet-State Population

The range of applications for fluorescent dyes in medicine and biology is increasing greatly. At the same time, the demands on the dyes are getting bigger. The dye molecules are often expected to emit maximum-intensity fluorescence. The primary requirement for this is naturally a high fluorescence quantum yield. Beyond this, it must be considered that fluorescent molecules located in a meta-stable triplet state are not able to emit fluorescent radiation. This represents a reduction of the intensity of the fluorescence, which is generally underestimated in its importance. The size of this perturbation can be grasped by knowledge of the quantum yield for occupation of the triplet state and its lifetime. Unfortunately these parameters are completely unknown, even for frequently used dyes. On investigating some medically and biologically relevant dyes, it is shown that the attainable fluorescence intensity could be reduced strongly by occupation of the triplet state.