- Specific and photostable rhodamine-based tracker for 3D video imaging of single acidic organelles
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Three-dimensional video imaging has emerged as an indispensable tool for real-time monitoring of dynamic acidic organelles. However, the limitation of video imaging is the absence of a specific stain for acidic organelle trackers. The aim of this work was to investigate the applicability of a potential acidic organelle tracker, Lyso-R, in three-dimensional video imaging in live cells. In a close examination of three differently designed rhodamine dyes, Lyso-R outperformed the other two with a suitable pKa value and higher membrane permeability. The uninterrupted fluorescence of Lyso-R towards macromolecules, e.g. lecithin and proteins, led to higher specificity and signal-to-background ratio than LysoTracker DND 189 and DND 99 for imaging acidic organelles. In addition, Lyso-R was photostable, and MTT assays confirmed its low toxicity towards cells. Inspired by these facts, three-dimensional tracking of a single acidic organelle in a live cell was obtained by staining with Lyso-R under confocal microscopy. The measurement of this organelle demonstrated that the distance change of the organelle centroid on XY plane was sharper than its depth change. The usage of Lyso-R was further extended with two-dimensional video imaging of acidic organelles during various cell metabolisms. All of these results demonstrate the potential applicability of Lyso-R as a three-dimensional imaging tracker of acidic organelles. the Partner Organisations 2014.
- Ye, Zhiwei,Xiao, Yi,Guo, Haiying,Wang, Chao
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- Synthesis of isomerically pure carboxylate- and sulfonate-substituted xanthene fluorophores
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Xanthene-based fluorophores such as fluorescein and rhodamine are typically prepared by acid-catalyzed condensation of the appropriate resorcinol or 3-aminophenol with phthalic anhydride. Condensation of substituted phthalic anhydride species results in functionalized fluorophores that are formed as mixed isomers. Crystallization approaches to isomer separation have been reported elsewhere for symmetric fluorescein carboxylates. We describe crystallization-based separation of protected fluorescein sulfonates and coupling conditions to form sulfonamides, precursors for carboxylate-substituted rhodamines, and precursors for asymmetrically substituted fluoresceins and rhodafluors.
- Woodroofe, Carolyn C.,Lim, Mi Hee,Bu, Weiming,Lippard, Stephen J.
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- Boronate ester cross-linked PVA hydrogels for the capture and H2O2-mediated release of active fluorophores
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A new set of PVA hydrogels were formed using the boronate ester fluorescent probe PF1 and the novel boronate fluorescent probe PT1 as the covalent crosslinkers. Treatment with aqueous H2O2 allowed triggered release of the fluorescent dye accompanied by complete dissolution of the hydrogel.
- Williams, George T.,Sedgwick, Adam C.,Sen, Sajal,Gwynne, Lauren,Gardiner, Jordan E.,Brewster, James T.,Hiscock, Jennifer R.,James, Tony D.,Jenkins, A. Toby A.,Sessler, Jonathan L.
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supporting information
p. 5516 - 5519
(2020/06/10)
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- Triarylmethane Fluorophores Resistant to Oxidative Photobluing
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Spectral stability of small-molecule fluorescent probes is required for correct interpretation and reproducibility of multicolor fluorescence imaging data, in particular under high (de)excitation light intensities of super-resolution imaging or in single-molecule applications. We propose a synthetic approach to a series of spectrally stable rhodamine fluorophores based on sequential Ru- and Cu-catalyzed transformations, evaluate their stability against photobleaching and photoconversion in the context of other fluorophores using chemometric analysis, and demonstrate chemical reactivity of fluorophore photoproducts. The substitution patterns providing the photoconversion-resistant triarylmethane fluorophores have been identified, and the applicability of nonbluing labels in live-cell STED nanoscopy is demonstrated.
- Butkevich, Alexey N.,Bossi, Mariano L.,Lukinavi?ius, Gra?vydas,Hell, Stefan W.
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supporting information
p. 981 - 989
(2019/01/23)
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- Ferric hydrogensulphate as a recyclable catalyst for the synthesis of fluorescein derivatives
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Polycondensation reactions of phenols with phthalic anhydride were carried out in the presence of ferric hydrogensulphate under melt conditions. The reactions proceeded in short reaction times by using a catalytic amount of Fe(HSO4)3 and the corresponding fluorescein derivatives were obtained in high yields. The simplicity, scale-up, along with the use of an inexpensive, non-toxic, recyclable catalyst of an environmentally benign nature, are other remarkable features of the procedure. The absorption and emission properties of these fluorescein derivatives were studied.
- Eshghi, Hossein,Mirzaie, Narges
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experimental part
p. 504 - 509
(2012/07/13)
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