There is still great need in better characterizing new targeted therapies in vivo, especially prior to clinical translation. In this regard, preclinical molecular imaging is a central tool in the targeted drug development pipeline. However, there is still a lack of imaging platforms that can enable to monitor target engagement, a key in vitro/in vivo parameter to assess the probability of success for a lead compound. Here, we will report on our methodologies to quantify target engagement via lifetime FRET in live subjects. These will include our recent instrumental developments, including large SPAD arrays and structured light strategies, as well as, software developments, including AI based data processing pipelines for 2D/3D imaging and unmixing.
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