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WIREs Nanomed Nanobiotechnol
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NIR fluorescent small molecules for intraoperative imaging

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Recent advances in bioimaging and nanomedicine have permitted the exploitation of molecular optical imaging in image‐guided surgery; however, the parameters mediating optimum performance of contrast agents are not yet precisely determined. To develop ideal contrast agents for image‐guided surgery, we need to consider the following criteria: (1) excitation and emission wavelengths in the near‐infrared (NIR) window, (2) optimized optical characteristics for high in vivo performance, (3) overcoming or harnessing biodistribution and clearance, and (4) reducing nonspecific uptake. The design considerations should be focused on optimizing the optical and physicochemical property criteria. Biodistribution and clearance should first be considered because they mediate the fate of a contrast agent in the body such as how long after intravenous injection a contrast agent reaches the peak signal‐to‐background ratio (SBR) and how long the signal lasts (retention). WIREs Nanomed Nanobiotechnol 2015, 7:828–838. doi: 10.1002/wnan.1337 This article is categorized under: Diagnostic Tools > In Vivo Nanodiagnostics and Imaging
(a) PET imaging. 100 µCi of scVEGF/Cu (top row) or inVEGF/Cu (bottom row) were injected into 4T1luc tumor‐bearing mice in the left axillary fat pads. Radioactivity for each resected organ was obtained 2 h post‐injection. (b) SPECT imaging. 100 µCi of scVEGF/Tc (left) or inVEGF/Tc (right) were injected into 4T1luc tumor‐bearing mice 1 h prior to imaging and resection. Arrows mark left mammary fat pad tumor; arrowheads and dotted boxes indicate nonspecific uptake. H, heart; L, liver; scale bar, 1 cm (Reprinted with permission from Ref . Copyright 2007 Nature Publishing Group).
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Biodistribution and clearance of ICG and ZW800‐1. ICG shows elevated nonspecific uptake in liver and duodenum, while ZW800‐1 represents ultralow background and renal clearance. Animals were housed in an AAALAC‐certified facility and were studied under the supervision of BIDMC IACUC in accordance with approved institutional protocols (#101‐2011 for rodents and #046‐2010 for pigs). NIR fluorophores were injected intravenously into ∼20 g CD‐1 mice (10 nmol), ∼250 g SD rats (50 nmol), and ∼35 kg Yorkshire pigs (1 µmol) 1 h prior to imaging. Shown are color and 800 nm NIR fluorescence images of surgically exposed organs taken by the FLARE intraoperative imaging system. NIR fluorescence images have identical exposure times and normalizations. Bl: bladder; BD: bile duct; Du: duodenum; In: intestine; Ki: kidneys; Li: liver; Sk: skin; St: stomach; Ur: ureter. Scale bars = 1 cm.
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Current state‐of‐the‐art NIR fluorophores used in image‐guided surgery. The chemical structures of three NIR fluorescent contrast agents and the corresponding optical properties. The negatively charged moieties are highlighted in red and the positively charged regions are designated in blue.
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Chemical structures of various classes of small molecule fluorophores that have been explored for biophotonic imaging, their general hydrophobicity maps, highlighted characteristics concerning their potential for in vivo performance and the overall optical properties concerning the general core structure.
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In vivo optical properties of injected fluorophores along with wavelength.
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