Long fluorescence lifetime triangulenium dyes in imaging and fluorescence polarization assay

The development of new fluorescent dyes-new fluorochromes-has a large potential to improve the established methods in enzymology, by empowering both detection capability and the scope of the individual method. Unfortunately, there are huge barriers when adopting new improved fluorescent dyes in established methods. The dyes have to be generally available, protocols for labeling and analysis must be in place, and the field has to be aware how the new improved dye can enhance their method of choice. In this chapter, we will address these issues for the triangulenium dyes. A class of dyes that has a long fluorescence lifetime and emission in the red. A unique combination that opens up new possibilities for the study of protein rotational motion, when developing fluorescence polarization (FP) assays, and for all time-resolved imaging or analysis platforms. To make these dyes generally available, the features of the long fluorescence lifetime triangulenium dyes are described and an optimized labelling protocol are reported.