A FRET-based ratiometric redox probe for detecting oxidative stress by confocal microscopy, FLIM and flow cytometry

Kaur, Amandeep; Haghighatbin, Mohammad A.; Hogan, Conor F.; New, Elizabeth J.

doi:10.1039/c5cc03394b

Cited by 66 publications

(68 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We have previously demonstrated that synthetic flavins can act as redox-reporting groups in sensors (28,29,46). Here, we have designed a class of flavin-rhodamine redox (FRR) sensors bearing a flavin redox-responsive group and a second fluorophore, rhodamine, which acts as a Förster resonance energy transfer (FRET) acceptor (Fig.…”

Section: Resultsmentioning

confidence: 99%

Studies of Hematopoietic Cell Differentiation with a Ratiometric and Reversible Sensor of Mitochondrial Reactive Oxygen Species

Kaur

Jankowska

Pilgrim

et al. 2016

Antioxidants & Redox Signaling

Self Cite

View full text Add to dashboard Cite

Aims: Chronic elevations in cellular redox state are known to result in the onset of various pathological conditions, but transient increases in reactive oxygen species (ROS)/reactive nitrogen species (RNS) are necessary for signal transduction and various physiological functions. There is a distinct lack of reversible fluorescent tools that can aid in studying and unraveling the roles of ROS/RNS in physiology and pathology by monitoring the variations in cellular ROS levels over time. In this work, we report the development of ratiometric fluorescent sensors that reversibly respond to changes in mitochondrial redox state. Results: Photophysical studies of the developed flavin-rhodamine redox sensors, flavin-rhodamine redox sensor 1 (FRR1) and flavin-rhodamine redox sensor 2 (FRR2), confirmed the reversible response of the probes upon reduction and re-oxidation over more than five cycles. The ratiometric output of FRR1 and FRR2 remained unaltered in the presence of other possible cellular interferants (metals and pH). Microscopy studies indicated clear mitochondrial localization of both probes, and FRR2 was shown to report the time-dependent increase of mitochondrial ROS levels after lipopolysaccharide stimulation in macrophages. Moreover, it was used to study the variations in mitochondrial redox state in mouse hematopoietic cells at different stages of embryonic development and maturation. Innovation: This study provides the first ratiometric and reversible probes for ROS, targeted to the mitochondria, which reveal variations in mitochondrial ROS levels at different stages of embryonic and adult blood cell production. Conclusions: Our results suggest that with their ratiometric and reversible outputs, FRR1 and FRR2 are valuable tools for the future study of oxidative stress and its implications in physiology and pathology. Antioxid. Redox Signal. 24, 667-679.

show abstract

Section: Resultsmentioning

confidence: 99%

Studies of Hematopoietic Cell Differentiation with a Ratiometric and Reversible Sensor of Mitochondrial Reactive Oxygen Species

Kaur

Jankowska

Pilgrim

et al. 2016

Antioxidants & Redox Signaling

Self Cite

View full text Add to dashboard Cite

show abstract

“…FCR1 [50] Global 820 TP /470, 520 520/470 6-fold increase HeLa -NAC and H 2 O 2 Angewandte Chemie Minireviews upon reversible reduction to the diamagnetic hydroxylamine or irreversible formation of alkoxyamine derivatives (Figure 2a). Since fluorescence is activated upon reduction, these probes are often referred to as profluorescent nitroxides,i n analogy to prodrugs.…”

Section: Nitroxide-based Probesmentioning

confidence: 98%

“…450/512 % 9-fold increase HeLa -Na 2 S 2 O 4 and H 2 O 2 NpFR1 [48] Global 405/5451 25-fold increase 3T3-L1 preadipocytes and differentiated adipoctyes -Glucose and H 2 O 2 NpFR2 [49] Global 488/5451 15-fold increase Mice bone marrow,thymus and spleen -H 2 O 2 and Mitotracker deep red. FCR1 [50] Global 820 TP /470, 520 520/470 6-fold increase…”

Section: Raw2647 Cells-h 2 O 2 E Po Kbr and Gstmentioning

confidence: 99%

“…[49] Both probes demonstrate greater than 100-fold decreases in fluorescence upon reduction, with excellent reversibility behavior with ar ange of oxidizing agents.N pFR1 was used to reveal the impact of glucose on oxidative stress in 3T3-L1 adipocytes,w hile NpFR2 was employed in flow cytometry studies to distinguish between cell populations with different mitochondrial oxidative capacities within the bone marrow,thymus,and spleen of mice.Asecond-generation ratiometric redox probe,F CR1 (Figure 7c), was also reported by our group,and consisted of ac oumarin/flavin, donor-acceptor FRET pair. [50] In the oxidized form, aF RET process results in green emission from the flavin acceptor,w hile upon reduction, energy transfer is inhibited, thus resulting in blue donor emission. In addition to fluorescence microscopy and flow cytometry, FCR1 was also shown to report on cellular oxidative capacity by fluorescence lifetime imaging microscopy (FLIM).…”

Section: Nicotinamides and Flavinsmentioning

confidence: 99%

See 1 more Smart Citation

Reversible Fluorescent Probes for Biological Redox States

2015

Self Cite

View full text Add to dashboard Cite

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.

show abstract

“… Reversible Redoxumwandlungen von a) Nicotinamid und b) der Flavineinheit. c) Ausgewählte Beispiele von Sonden, die auf der Basis dieser redoxresponsiven Gruppen entwickelt wurden …”

Section: Nicotinamide Und Flavineunclassified

Reversible Fluoreszenzsonden für biologische Redoxzustände

2015

Self Cite

View full text Add to dashboard Cite

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.

show abstract

A FRET-based ratiometric redox probe for detecting oxidative stress by confocal microscopy, FLIM and flow cytometry

Cited by 66 publications

References 29 publications

Studies of Hematopoietic Cell Differentiation with a Ratiometric and Reversible Sensor of Mitochondrial Reactive Oxygen Species

Studies of Hematopoietic Cell Differentiation with a Ratiometric and Reversible Sensor of Mitochondrial Reactive Oxygen Species

Reversible Fluorescent Probes for Biological Redox States

Reversible Fluoreszenzsonden für biologische Redoxzustände

Contact Info

Product

Resources

About