Using a TEMPO-based fluorescent probe for monitoring oxidative stress in living cells

Liu, Yang; Zhu, Mingyang; Xu, Jingji; Zhang, Hong; Tian, Mei

doi:10.1039/c1an15417f

Cited by 17 publications

(14 citation statements)

References 21 publications

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“…[19] This probe could detect H 2 O 2 -induced oxidative stress in Hep2G hepatocellular carcinoma cells,and was stable to the hydrolytic activity of trypsin, for at ime period of 60 minutes,invitro. [20] While further red-shifting of excitation and emission wavelengths was achieved by conjugating the TEMPO radical to either as tyryl BODIPY,N ile Red, or ar uthenium polypyridyl complex, biological application of these probes was precluded by low quantum yields (0.002 to 0.050) and relatively small dynamic ranges (two-to eightfold changes). [25] In another study,awater-insoluble silicato-phtalocyaninebased profluorescent nitroxide (R2c), with emission at l = 700 nm, was encapsulated in liposomes and used to quantify ascorbic acid levels in HeLa cells.…”

Section: Hela -Nac and H 2 Omentioning

confidence: 99%

“…One of the probes that successfully addressed the limited applicability of PFNs with short wavelength excitations in biological studies is TMB (Figure b), a green‐emitting BODIPY fluorophore decorated with a nitroxide moiety, with a sixfold increase in emission upon reduction . This probe could detect H 2 O 2 ‐induced oxidative stress in Hep2G hepatocellular carcinoma cells, and was stable to the hydrolytic activity of trypsin, for a time period of 60 minutes, in vitro …”

Section: Nitroxide‐based Probesmentioning

confidence: 99%

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Reversible Fluorescent Probes for Biological Redox States

2015

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The redox chemistry of the cell is key to its function and health, and the development of chemical tools to study redox biology is important. While fluxes in oxidative state are essential for healthy cell function, a chronically elevated oxidative capacity is linked to disease. It is therefore essential that probes of biological redox states distinguish between these two conditions by the reversible sensing of changes over time. In this review, we discuss the current progress towards such probes, and identify key directions for future research in this nascent field of vital biological interest.

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Section: Hela -Nac and H 2 Omentioning

confidence: 99%

Section: Nitroxide‐based Probesmentioning

confidence: 99%

Reversible Fluorescent Probes for Biological Redox States

2015

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“…Eine der Sonden, mit denen das Problem der begrenzten Anwendbarkeit von PFNs mit Anregungen im kurzwelligen Bereich in biologischen Untersuchungen gemeistert werden konnte, ist TMB (Abbildung b), ein grün emittierender BODIPY‐Fluorophor mit einem Nitroxidbaustein, der bei Reduktion eine sechsfache Zunahme der Emission zeigt . Diese Sonde konnte H 2 O 2 ‐induzierten oxidativen Stress in den hepatozellulären Hep2G‐Karzinomzellen detektieren und war in vitro 60 min stabil gegen die hydrolytische Aktivität von Trypsin …”

Section: Sonden Auf Nitroxidbasisunclassified

Reversible Fluoreszenzsonden für biologische Redoxzustände

2015

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Die Redoxchemie der Zelle ist für deren Funktionstüchtigkeit und Gesundheit von entscheidender Bedeutung. Dementsprechend ist es wichtig, chemische Hilfsmittel für die Untersuchung der Redoxbiologie zu entwickeln. Während Veränderungen im oxidativen Zustand für eine gesunde Zellfunktion wesentlich sind, wird eine chronisch erhöhte oxidative Kapazität mit Krankheiten in Verbindung gebracht. Es ist daher entscheidend, dass Sonden biologischer Redoxzustände zwischen diesen beiden Bedingungen unterscheiden können, indem sie Veränderungen reversibel über die Zeit erfassen. In diesem Kurzaufsatz diskutieren wir die gegenwärtigen Fortschritte bei solchen Sonden und ermitteln die Hauptrichtungen für zukünftige Forschungen auf diesem aufstrebenden Gebiet, das von grundlegendem biologischem Interesse ist.

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“…21 The reaction principle of the TEMPO-based fluorescent probe was detailed in Scheme 1. 23 The newly synthesized probe was used to evaluate the changes of redox status of L02 cells which was pretreated by H 2 O 2 . Since the cell membrane is permeable to the new dye, the incubation time of 30 min was used to guarantee complete derivatization of intracellular reducing species.…”

Section: Considerations In Redox-sensitive Fluorescent Probe For Livimentioning

confidence: 99%

Evaluating intracellular redox status in L02 cells on microchip

Xu¹,

Cai²

2013

J. Braz. Chem. Soc.

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Um novo dispositivo microfluídico integrado com introdução contínua de células individuais e detecção de espécies redutoras intracelulares por detector de fluorescência induzida por laser (LIF) é relatado. Um canal central de amostragem foi desenhado sendo flanqueado por dois outros canais auxiliares. As espécies intracelulares redutoras foram derivadas por uma sonda fluorescente baseada na recém-sintetizada 2,2,6,6-tetrametil-piperidina-1-oxilo (TEMPO). As células marcadas foram hidrodinamicamente focadas pelas correntes da bainha de fluxo e sequencialmente introduzidas no canal de amostragem sob pressão hidrostática gerada por ajuste dos níveis de líquido nos reservatórios. O nível de redução intracelular em células vivas intactas L02 foi detectada por LIF sem citólise. Após estimulação com 100 mmol L-1 de peróxido de hidrogênio, durante 30 minutos, o nível de redução intracelular diminuiu em resposta ao estresse oxidativo. Foi obtida uma taxa de transferência de 41-44 células min-1. A novel microfluidic device integrated with continuous introduction of individual cells and detection of intracellular reducing species by laser induced fluorescence (LIF) detector is reported. A single channel with one sheath-flow channel located on each side of the sampling channel was designed. The intracellular reducing species were derivatized by a newly synthesized 2,2,6,6-tetramethyl-piperidine-1-oxyl (TEMPO)-based fluorescent probe. The labeled cells were hydrodynamically focused by sheath-flow streams and sequentially introduced into the sampling channel under hydrostatic pressure generated by adjusting liquid levels in the reservoirs. Intracellular reducing level in intact living L02 cells was detected by LIF without cytolysis. Upon stimulation with 100 μmol L-1 hydrogen peroxide for 30 min, the intracellular reducing level decreased in response to oxidative stress. A throughput of 41-44 cells min-1 was obtained.

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Using a TEMPO-based fluorescent probe for monitoring oxidative stress in living cells

Cited by 17 publications

References 21 publications

Reversible Fluorescent Probes for Biological Redox States

Reversible Fluorescent Probes for Biological Redox States

Reversible Fluoreszenzsonden für biologische Redoxzustände

Evaluating intracellular redox status in L02 cells on microchip

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