Synthesis of Through-bond Energy Transfer Cassettes and Their Encapsulation in Silica and Calcium Phosphate Nanoparticles

Water-soluble fluorescent probes with emission in the 600-800 nm region have significant potential in biological applications such as cell imaging. Most fluorescent probes however suffer from limited fluorescence brightness in aqueous media due to aggregation and self-quenching. Their photostability in animal models for an extended period of time is also a concern. One way of improving their photophysical properties is to encapsulate them in a protective matrix to form fluorescent nanoparticles.

We have synthesized a set of six through-bond energy transfer cassettes which emit in the 600-800 nm region with Fluorescein or BODIPY as donor and benzophenoxazine dye Nile Red or cyanine dye Cy5 as acceptor. Their photophysical properties in organic and aqueous media were evaluated. Some of these cassettes were encapsulated in silica or calcium phosphate nanoparticles (20 nm in diameter) to improve their solubility and photophysical properties in aqueous media. We also synthesized some water-soluble benzophenoxazine based fluorophores and the impact of different water-soluble groups on their emission characteristics in aqueous media was studied. Selected fluorophores were used for in vitro cellular imaging studies.