A Dual Fluorescence-Spin Label Probe for Visualization and Quantification of Target Molecules in Tissue by Multiplexed FLIM-EPR Spectroscopy
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Date
2021-06-25Author
Dong, Pin
Stellmacher, Johannes
Bouchet, Lydia M.
Nieke, Marius
Kumar, Amit
Osorio Blanco, Ernesto Rafael
Nagel, Gregor
Lohan, Silke B.
Teutloff, Christian
Patzelt, Alexa
Schäfer-Korting, Monika
Calderón, Marcelo
Meinke, Martina C.
Alexiev, Ulrike
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Angewandte Chemie International Edition 60(27) : 14938-14944 (2021)
Abstract
Simultaneous visualization and concentration quantification of molecules in biological tissue is an important though challenging goal. The advantages of fluorescence lifetime imaging microscopy (FLIM) for visualization, and electron paramagnetic resonance (EPR) spectroscopy for quantification are complementary. Their combination in a multiplexed approach promises a successful but ambitious strategy because of spin label-mediated fluorescence quenching. Here, we solved this problem and present the molecular design of a dual label (DL) compound comprising a highly fluorescent dye together with an EPR spin probe, which also renders the fluorescence lifetime to be concentration sensitive. The DL can easily be coupled to the biomolecule of choice, enabling in vivo and in vitro applications. This novel approach paves the way for elegant studies ranging from fundamental biological investigations to preclinical drug research, as shown in proof-of-principle penetration experiments in human skin ex vivo.