Optical methods

Molecular imaging requires the highest possible signal-to-noise ratios (SNRs) at the target of interest. In order to maximize the SNR for optical imaging techniques, various strategies have been developed to design fluorescent probes that can be activated, for example, by proteolytic degradation. Generally speaking, these probes are quenched in their native state-e.g., by fluorescence resonance energy transfer (FRET)-and dequenched after cleavage or hybridization, which is associated with a strong fluorescence signal increase. Different strategies of fluorescence signal amplification ranging from large and small protease-sensing molecules to oligonucleotide-sensing and nanoparticle-based probes are presented in this chapter.