12 research outputs found

    Influence of polymer composition on the sensitivity towards nitrite and nitric oxide of colorimetric disposable test strips

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    The influence of polymer composition on the sensitivity towards nitrite (NO 2 − ) and nitric oxide (NO) of a series of 19 polymeric hydrogel films has been stud- ied. The polymers, based on the hydrophilic monomer 2- hydroxyethylmethacrylate (HEMA), are able to encapsu- late the colorimetric indicator 1,2-diaminoanthraquinone (DAQ) and to respond to NO 2 − and NO by visual chang- es. In the case of nitrite, the calculated limits of detection (LOD) for two of the polymeric sensors (10 μ M) are very close to the sensitivity estimated for free DAQ in solution (LOD 5 μ M), but with the advantage of a solid supported sensor with the format of a disposable test- strip made with affordable starting chemicals. The re- sults are interpreted taking into account the nature and proportions of monomers and cross-linkers used for the synthesis of polymers. Key factors for obtaining sensi- tive materials are the hydro philic character of the film along with the utilization of low levels of cross-linker and the use of an acidic monomer, like acrylic acid, as a building block.Financial support from the Spanish MINECO (CTQ2015-68429-R) and Fundació Caixa Castelló-UJI (P1·1B2015-76) is acknowledged. V. F. thanks the financial support from UJI (predoctoral fellowship). We thank SCIC/UJI for technical assistance

    A photoinduced electron transfer-based nanoprobe as a marker of acidic organelles in mammalian cells

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    Photoinduced electron transfer (PET)-based molecular probes have been successfully used for the intracellular imaging of the pH of acidic organelles. In this study, we describe the synthesis and characterization of a novel PET-based pH nanoprobe and its biological application for the signaling of acidic organelles in mammalian cells. A fluorescent ligand sensitive to pH via the PET mechanism that incorporates a thiolated moiety was synthesized and used to stabilize gold nanoparticles (2.4?±?0.6 nm), yielding a PET-based nanoprobe. The PET nanoprobe was unambiguously characterized by transmission electron microscopy, proton nuclear magnetic resonance, Fourier transform infrared, ultraviolet-visible absorption, and steady-state/time-resolved fluorescence spectroscopies which confirmed the functionalization of the gold nanoparticles with the PET-based ligand. Following a classic PET behavior, the fluorescence emission of the PET-based nanoprobe was quenched in alkaline conditions and enhanced in an acidic environment. The PET-based nanoprobe was used for the intracellular imaging of acidic environments within Chinese hamster ovary cells by confocal laser scanning microscopy. The internalization of the nanoparticles by the cells was confirmed by confocal fluorescence images and also by recording the fluorescence emission spectra of the intracellular PET-based nanoprobe from within the cells. Co-localization experiments using a marker of acidic organelles, LysoTracker Red DND-99, and a marker of autophagosomes, GFP-LC3, confirm that the PET-based nanoprobe acts as marker of acidic organelles and autophagosomes within mammalian cells
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