Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 60, Issue 27, Pages 14938-14944Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202012852
Keywords
3-carboxy-proxyl (PCA); fluorescence; multiplexed FLIM– EPR spectroscopy; Rhodamine B; spin labels
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Funding
- German Research Foundation
- DFG
- MINECO project [RTI2018-099227-B-I00]
- Dahlem Research School
- China Scholarship Council
- Projekt DEAL
- IKERBASQUE-Basque Foundation for Science
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The combination of FLIM and EPR in a multiplexed approach using DL compound enables simultaneous visualization and concentration quantification of molecules in biological tissue, paving the way for elegant studies from fundamental biological investigations to preclinical drug research. This novel approach has been successfully demonstrated in proof-of-principle penetration experiments in human skin ex vivo.
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.
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