Defining a Polymethine Dye for Fluorescence Anisotropy Applications in the Near‐Infrared Spectral Range

Gustafson, Tiffany P.; Cao, Qian; Achilefu, Samuel; Berezin, Mikhail Y.

doi:10.1002/cphc.201100916

Cited by 21 publications

(32 citation statements)

References 58 publications

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“…Fluorescence anisotropy was conducted in L-format with automated Glan-Thompson polarizing prisms controlled by FluorEssence software (Horiba) as described earlier. 47 …”

Section: Methodsmentioning

confidence: 99%

Blood triggered rapid release porous nanocapsules

et al. 2013

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Rapid-release drug delivery systems present a new paradigm in emergency care treatments. Such systems combine a long shelf life with the ability to provide a significant dose of the drug to the bloodstream in the shortest period of time. Until now, development of delivery formulations has concentrated on slow release systems to ensure a steady concentration of the drug. To address the need for quick release system, we created hollow polyacrylate nanocapsules with nanometer-thin porous walls. Burst release occurs upon interaction with blood components that leads to escape of the cargo. The likely mechanism of release involves a conformational change of the polymer shell caused by binding albumin. To demonstrate this concept, a near-infrared fluorescent dye indocyanine green (ICG) was incorporated inside the nanocapsules. ICG-loaded nanocapsules demonstrated remarkable shelf life in aqueous buffers with no release of ICG for twelve months. Rapid release of the dye was demonstrated first in vitro using albumin solution and serum. SEM and light scattering analysis demonstrated the retention of the nanocapsule architecture after the release of the dye upon contact with albumin. In vivo studies using fluorescence lifetime imaging confirmed quick discharge of ICG from the nanocapsules following intravenous injection.

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“…Fluorescence anisotropy was conducted in L-format with automated Glan-Thompson polarizing prisms controlled by FluorEssence software (Horiba) as described earlier. 47 …”

Section: Methodsmentioning

confidence: 99%

Blood triggered rapid release porous nanocapsules

et al. 2013

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“…Dyes 1,1′,3,3,3′,3′-hexamethylindotricarbocyanine (HITC) was purchased from Exciton, IRDye800CW from LI-COR Biosciences, ADS740WS was obtained from American Dye Source, LS851 was prepared in the lab according to a standard cyanine synthesis procedure [25]. Reduced hydrocyanines HITC-H, IRdye800CW-H, ADS740WS–H, and LS851-H were prepared according to a previously published procedure [20].…”

Section: Methodsmentioning

confidence: 99%

Imaging of radicals following injury or acute stress in peripheral nerves with activatable fluorescent probes

Zhou

Yan

et al. 2016

Free Radical Biology and Medicine

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Peripheral nerve injury evokes a complex cascade of chemical reactions including generation of molecular radicals. Conversely, the reactions within nerve induced by stress are difficult to directly detect or measure to establish causality. Monitoring these reactions in vivo would enable deeper understanding of the nature of the injury and healing processes. Here, we utilized near-infrared fluorescence molecular probes delivered via intra-neural injection technique to enable live, in vivo imaging of tissue response associated with nerve injury and stress. These initially quenched fluorescent probes featured specific sensitivity to hydroxyl radicals and become fluorescent upon encountering reactive oxygen species (ROS). Intraneurally delivered probes demonstrated rapid activation in injured rat sciatic nerve but minimal activation in normal, uninjured nerve. In addition, these probes reported activation within sciatic nerves of living rats after a stress caused by a pinprick stimulus to the abdomen. This imaging approach was more sensitive to detecting changes within nerves due to the induced stress than other techniques to evaluate cellular and molecular changes. Specifically, neither histological analysis of the sciatic nerves, nor the expression of pain and stress associated genes in dorsal root ganglia could provide statistically significant differences between the control and stressed groups. Overall, the results demonstrate a novel imaging approach to measure ROS in addition to the impact of ROS within nerve in live animals.

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“…31 For oblate molecules such as cyanine dyes, especially NIR cyanine dyes (see below), this lack of symmetry leads to larger initial anisotropies ( r min > 0.17) that limit the dynamic range. 34 Large values for r min is the major limitation to the use of near infrared (NIR) dyes, for example, in drug assays. 35 …”

Section: Steady-state Fluorescence Anisotropymentioning

confidence: 99%

“…This leads to incomplete immobilization of the fluorescent probe on the macromolecule of interest and, therefore, to higher local mobility of the fluorophore via the propeller effect. To address these problems, Gustafson et al 34 evaluated a number of NIR dyes with the goal of identifying a dye for FA in the NIR spectral range. The study identified a dye with low anisotropy in the free (non-bound) state and demonstrated the potential of NIR anisotropy in analytical applications.…”

Section: Steady-state Fluorescence Anisotropymentioning

confidence: 99%

“…86, 87 FA has also been utilized to verify the conjugation efficiency of fluorophores to nanoparticles and as a quality control method to verify fluorescent labeling in diagnostic probes. 34, 88 Using FA, Sahoo et al 89 confirmed the labeling of biocompatible magnetic nanoparticles with Rhodamine 110. In water this free rhodamine exhibited almost no FA ( r ~ 0), but FA increased up to 0.4 when the dye was bound to nanoparticles.…”

Section: Novel Applications: Biologics Nanotechnology and Imaginmentioning

confidence: 99%

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Fluorescence anisotropy (polarization): from drug screening to precision medicine

Zhang

Berezin

2015

Expert Opinion on Drug Discovery

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Introduction Fluorescence anisotropy (FA) is one of the major established methods accepted by industry and regulatory agencies for understanding the mechanisms of drug action and selecting drug candidates utilizing a high-throughput format. Areas covered This review covers the basics of FA and complementary methods, such as fluorescence lifetime anisotropy and their roles in the drug discovery process. The authors highlight the factors affecting FA readouts, fluorophore selection, and instrumentation. Furthermore, the authors describe the recent development of a successful, commercially valuable FA assay for Long QT syndrome drug toxicity to illustrate the role that FA can play in the early stages of drug discovery. Expert opinion Despite the success in drug discovery, the FA-based technique experiences competitive pressure from other homogeneous assays. That being said, FA is an established yet rapidly developing technique, recognized by academic institutions, the pharmaceutical industry, and regulatory agencies across the globe. The technical problems encountered in working with small molecules in homogeneous assays are largely solved, and new challenges come from more complex biological molecules and nanoparticles. With that, FA will remain one of the major work-horse techniques leading to precision (personalized) medicine.

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Defining a Polymethine Dye for Fluorescence Anisotropy Applications in the Near‐Infrared Spectral Range

Cited by 21 publications

References 58 publications

Blood triggered rapid release porous nanocapsules

Blood triggered rapid release porous nanocapsules

Imaging of radicals following injury or acute stress in peripheral nerves with activatable fluorescent probes

Fluorescence anisotropy (polarization): from drug screening to precision medicine

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