In vitro analysis of cell metabolism using a long-decay pH-sensitive lanthanide probe and extracellular acidification assay

Hynes, James T.; O’Riordan, Tomás C.; Zhdanov, Alexander V.; Uray, Georg; Will, Yvonne; Papkovsky, Dmitri B.

doi:10.1016/j.ab.2009.04.016

Cited by 55 publications

(59 citation statements)

References 34 publications

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“…[22] These complexes are widely applied as labels in various luminescence assays both as free dyes bearing a functional group for conjugation and in the form of dye-doped nanoparticles, [22,23] but application in optical sensors and analyte-sensitive probes is comparably rare. [24][25][26][27] For example, luminescent Eu(III) complexes were applied as optical temperature probes, [27][28][29][30][31] pH indicators, [32][33][34][35][36] fluoroionophores for bicarbonate, [37,38] citrate [39] and lactate [40] and as hydrogen peroxide [41] and nitrogen monooxide probes. [42] Several Eu(III) complexes were applied as luminescent oxygen indicators.…”

Section: Introductionmentioning

confidence: 99%

Exceptional Oxygen Sensing Properties of New Blue Light‐Excitable Highly Luminescent Europium(III) and Gadolinium(III) Complexes

Borisov

Fischer

Saf

et al. 2014

Adv Funct Materials

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New europium(III) and gadolinium(III) complexes bearing 8-hydroxyphenalenone antenna combine efficient absorption in the blue part of the spectrum and strong emission in polymers at room temperature. The Eu(III) complexes show characteristic red luminescence whereas the Gd(III) dyes are strongly phosphorescent. The luminescence quantum yields are about 20% for the Eu(III) complexes and 50% for the Gd(III) dyes. In contrast to most state-of-the-art Eu(III) complexes the new dyes are quenched very efficiently by molecular oxygen. The luminescence decay times of the Gd(III) complexes exceed 1 ms which ensures exceptional sensitivity even in polymers of moderate oxygen permeability. These sensors are particularly suitable for trace oxygen sensing and may be good substitutes for Pd(II) porphyrins. The photophysical and sensing properties can be tuned by varying the nature of the fourth ligand. The narrow-band emission of the Eu(III) allows efficient elimination of the background light and autofluorescence and is also very attractive for use e.g. in multi-analyte sensors. The highly photostable indicators incorporated in nanoparticles are promising for imaging applications. Due to the straightforward preparation and low cost of starting materials the new dyes represent a promising alternative to the state-ofthe-art oxygen indicators particularly for such applications as e.g. food packaging.

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Section: Introductionmentioning

confidence: 99%

Exceptional Oxygen Sensing Properties of New Blue Light‐Excitable Highly Luminescent Europium(III) and Gadolinium(III) Complexes

Borisov

Fischer

Saf

et al. 2014

Adv Funct Materials

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“…It can, however, be harnessed for screening whereby OCR is used as the primary indicator and ECA is used as a confirmatory parameter in both 96 and 384 well plate format. In such case, true mitochondria inhibitors will cause a decrease in O 2 consumption and an accompanying increase in ECA while general cell toxicity will cause a decrease in both parameters [18] as no compensation will be possible. Uncouplers would be expected to increase in both ECA and OCR.…”

Section: Analysis Of Drug-induced Mitochondrial Dysfunction By Parallmentioning

confidence: 98%

“…The automated Extracellular Flux Analyser can accurately measure cellular OCR and ECA in a special microplate with builtin solid-state O 2 and pH sensors [43]; however, it is not able to differentiate between glycolytic and non-glycolytic contribution to the ECA. The more flexible platform based on the long decay emitting O 2 and pH sensitive probes overcomes this limitation [18,44]. Using standard microtiter plates and TR-F plate reader detection, it can measure lactate in unsealed (L-ECA, CO 2 escapes from the medium) and lactate plus CO 2 in sealed samples (total, T-ECA), thus discriminating between the glycolytic and non-glycolytic ECA components.…”

Section: Multi-parametric Assessment Of 'Cell Energy Budget'mentioning

confidence: 99%

“…The ECA can be followed using fluorescent pH-sensitive probes which, when spectrally compatible, facilitate true multiplexing where both probes are deployed together thereby reporting on both oxidative and glycolytic metabolism in the same test well. For example, the pH-sensitive probe pH-Xtra, which comprises a long decay narrow band emitting Eu(III)-chelate, can be multiplexed with Ptporphyrin-based MitoXpress Ò ecO 2 probe (produced by Luxcel Biosciences, Ireland), to achieve simultaneous readout of O 2 and H + concentrations within the same sample with practically no cross-sensitivity [18]. pH-Xtra probe also changes its emission lifetime as a function of pH, so that both probes can be measured in RLD mode on standard TR-F readers.…”

Section: Analysis Of Drug-induced Mitochondrial Dysfunction By Parallmentioning

confidence: 99%

“…Such instruments are produced by a number of vendors (e.g., FLUOstar and POLARStar Omega Ò instruments from BMG Labtech GmbH, Germany and the Victor Ò family from PerkinElmer, USA), they are available in many life sciences labs and can be used without major modifications with several existing Pt-porphyin-based O 2 sensing probes (also produced commercially). These instruments also allow for multiplexed and parallel use of O 2 probes with other compatible probes and assays [18].…”

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confidence: 99%

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O2 Analysis on a Fluorescence Spectrometer or Plate Reader

Zhdanov

Hynes

Dmitriev

2012

Phosphorescent Oxygen-Sensitive Probes

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In this chapter, the use of Pt-porphyrin-based extracellular and intracellular O 2 sensing probes (ecO 2 and icO 2 ) on a TR-F plate reader format is described and critically assessed. The principles underpinning extracellular measurement are outlined and the assessment of biological oxygen consumption in prokaryotic cells, eukaryotic cells and small organisms is outlined along with a description of how such measurements can contribute to the development of a detailed picture of cell metabolism. The use of a mathematical model describing the distribution of local O 2 gradients within biological samples is also described. Finally, the principles of icO 2 sensing are discussed and some short case studies are provided for demonstrating the utility of icO 2 probes for the monitoring of cellular function and metabolic perturbations and how such measurements can be allied to other bioenergetic markers to generate a more complete picture of metabolic status.

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Synthesis and Evaluation of Europium Complexes that Switch on Luminescence in Lysosomes of Living Cells

Starck

Fradgley

Pál

et al. 2020

Chemistry A European J

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A set of four luminescent EuIII complexes bearing an extended aryl‐alkynylpyridine chromophore has been studied, showing very different pH‐dependent behaviour in their absorption and emission spectral response. For two complexes with pKa values of 6.45 and 6.20 in protein‐containing solution, the emission lifetime increases very significantly following protonation. By varying the gate time during signal acquisition, the ‘switch‐on’ intensity ratio could be optimised, and enhancement factors of between 250 to 1330 were measured between pH 8 and 4. The best‐behaved probe showed no significant emission dependence on the concentration of endogenous cations, reductants, and serum albumin. It was examined in live‐cell imaging studies to monitor time‐dependent lysosomal acidification, for which the increase in observed image brightness due to acidification was a factor of 50 in NIH‐3T3 cells.

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In vitro analysis of cell metabolism using a long-decay pH-sensitive lanthanide probe and extracellular acidification assay

Cited by 55 publications

References 34 publications

Exceptional Oxygen Sensing Properties of New Blue Light‐Excitable Highly Luminescent Europium(III) and Gadolinium(III) Complexes

Exceptional Oxygen Sensing Properties of New Blue Light‐Excitable Highly Luminescent Europium(III) and Gadolinium(III) Complexes

O2 Analysis on a Fluorescence Spectrometer or Plate Reader

Synthesis and Evaluation of Europium Complexes that Switch on Luminescence in Lysosomes of Living Cells

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