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A Dual Fluorescence–Spin Label Probe for Visualization and Quantification of Target Molecules in Tissue by Multiplexed FLIM–EPR Spectroscopy
Author(s) -
Dong Pin,
Stellmacher Johannes,
Bouchet Lydia M.,
Nieke Marius,
Kumar Amit,
OsorioBlanco Ernesto R.,
Nagel Gregor,
Lohan Silke B.,
Teutloff Christian,
Patzelt Alexa,
SchäferKorting Monika,
Calderón Marcelo,
Meinke Martina C.,
Alexiev Ulrike
Publication year - 2021
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.202012852
Subject(s) - fluorescence , electron paramagnetic resonance , fluorescence cross correlation spectroscopy , molecule , multiplexing , nuclear magnetic resonance , spectroscopy , fluorescence spectroscopy , materials science , chemistry , visualization , analytical chemistry (journal) , chromatography , physics , computer science , optics , data mining , telecommunications , organic chemistry , quantum mechanics
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.