Insights into the etiology of stroke and myocardial infarction suggest that rupture of unstable atherosclerotic plaque is the precipitating event. Clinicians lack tools to detect lesion instability early enough to intervene, and are often left to manage patients empirically, or worse, after plaque rupture. Noninvasive imaging of the molecular events signaling prerupture plaque progression has the potential to reduce the morbidity and mortality associated with myocardial infarction and stroke by allowing early intervention. Here, we demonstrate proof-of-principle in vivo molecular imaging of C-type natriuretic peptide receptor in focal atherosclerotic lesions in the femoral arteries of New Zealand white rabbits using a custom built fiber-based, fluorescence molecular tomography (FMT) system. Longitudinal imaging showed changes in the fluorescence signal intensity as the plaque progressed in the air-desiccated vessel compared to the uninjured vessel, which was validated by ex vivo tissue studies. In summary, we demonstrate the potential of FMT for noninvasive detection of molecular events leading to unstable lesions heralding plaque rupture.
Fluorescence quenching properties of copper(II) ions have been used for designing Cu(II) sensitive fluorescent molecular probes. In this paper, we demonstrate that static quenching plays a key role in free Cu(II)-mediated fluorescence quenching of a near infrared (NIR) fluorescent dye cypate. The Stern-Volmer quenching constant was calculated to be KSV = 970,000 M-1 in 25 mM MES buffer, pH 6.5 at room temperature. We synthesized LS835, a compound containing cypate attached covalently to chelated Cu(II) to study fluorescence quenching by chelated Cu(II). The fluorescence quenching mechanism of chelated Cu(II) is predominantly dynamic or collisional quenching. The quenching efficiency of chelated Cu(II) was calculated to be 58% ± 6% in dimethylsulfoxide at room temperature. Future work will involve further characterization of the mechanism of NIR fluorescence quenching by Cu(II) and testing its reversibility for potential applications in designing fluorophore-quencher based molecular probes for biological imaging.
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