A Water-Soluble Luminescence Oxygen Sensor

Castellano, Felix N.; Lakowicz, Joseph R.

doi:10.1562/0031-8655(1998)067<0179:awslos>2.3.co;2

Cited by 84 publications

(33 citation statements)

References 20 publications

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“…Millipore FHUP Fluoropore™ Membrane Filter was bought from EMD Millipore Corporation (Billerica, MA, United States). (Ru[dpp(SO 3 Na) 2 3 )Cl 2 ) was synthesized in lab following to published results [40]. …”

Section: Methodsmentioning

confidence: 99%

Fast responsive, optical trace level ammonia sensor for environmental monitoring

Abel

Ungerböck

Klimant

et al. 2012

Chemistry Central Journal

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BackgroundAmmonia is a ubiquitous chemical substance which is created in technical and biological processes and harmful to many different organisms. One specific problem is the toxicity of ammonia in fish at levels of 25 μg/l - a very common issue in today’s aqua culture. In this study we report a development of a fast responsive, optical ammonia sensor for trace concentrations.ResultsDifferent hydrogels have been investigated as host polymers for a pH based sensing mechanism based on fluorescent dyes. A porous hydrophobic fluoropolymer membrane was used as an ion barrier cover layer to achieve a good ammonia permeability. The sensor’s sensitivity towards ammonia as well as crosssensitivity towards pH-value and salinity, and the temperature dependency have been determined. Two different methods to reference fluorescence signals have been employed to eliminate intensity-based measurement drawbacks.ConclusionThe presented sensor features high sensitivity and a fast response even at concentrations near 1 ppb. No cross sensitivity towards pH and salinity could be observed and temperature dependency was determined as compensateable. Both referencing approaches prove themselves to be able to provide a simple use of the sensor for in-field applications.

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Section: Methodsmentioning

confidence: 99%

Fast responsive, optical trace level ammonia sensor for environmental monitoring

Abel

Ungerböck

Klimant

et al. 2012

Chemistry Central Journal

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“…Measurements of oxygen consumption of L6 cells were performed by using Ru[dpp(SO 3 Na) 2 ] 3 (RU), a water soluble oxygen sensor. The probe was synthesized as␣described previously [32]. The principle of measurement was based on fluorescence quenching of the sensor by oxygen dissolved in the reaction medium.…”

Section: Methodsmentioning

confidence: 99%

Uncoupling of protein‐3 induces an uncontrolled uncoupling of mitochondria after expression in muscle derived L6 cells

Guerini

Prati

Desai

et al. 2002

European Journal of Biochemistry

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The uncoupling proteins (UCPs) are thought to uncouple oxidative phosphorylation in the mitochondria and thus generate heat. One of the UCP isoforms, UCP3, is abundantly expressed in skeletal muscle, the major thermogenic tissue in humans. UCP3 has been overexpressed at high levels in yeast systems, where it leads to the uncoupling of cell respiration, suggesting that UCP3 may indeed be capable of dissipating the mitochondrial proton gradient. This effect, however, was recently shown to be a consequence of the high level of expression and incorrect folding of the protein and not to its intrinsic uncoupling activity. In the present study, we investigated the properties of UCP3 overexpressed in a relevant mammalian host system such as the rat myoblast L6 cell line. UCP3 was expressed in relatively low levels (< 1 lgAEmg )1 membrane protein) with the help of an adenovirus vector. Immunofluorescence microscopy of transduced L6 cells showed that UCP3 was expressed in more than 90% of the cells and that its staining pattern was characteristic for mitochondrial localization. The oxygen consumption of L6 cells under nonphosphorylating conditions increased concomitantly with the levels of UCP3 expression. However, uncoupling was associated with an inhibition of the maximal respiratory capacity of mitochondria and was not affected by purine nucleotides and free fatty acids. Moreover, recombinant UCP3 was resistant to Triton X-100 extraction under conditions that fully solubilize membrane bound proteins.Thus, UCP3 can be uniformly overexpressed in the mitochondria of a relevant muscle-derived cell line resulting in the expected increase of mitochondrial uncoupling. However, our data suggest that the protein is present in an incompetent conformation.

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“…They can be used within a broad range of working concentrations from about 0.1 nM upwards. In contrast, oxygen probes on the basis of ruthenium(II) complexes, having lifetimes of only a few microseconds and less, 18 are not suitable for time-resolved measurements on most commercial time-resolved fluorescence plate readers.…”

Section: Optical and Quenching Properties Of Ptcp-bsa Conjugatesmentioning

confidence: 99%

“…However, their suitability to cell viability assays and HTS applications is questionable due to potential problems with reagent standardization, selfquenching at high concentrations, binding to cells and sample components (e.g., proteins), and phototoxic effects. Water-soluble oxygen probes based on fluorescent complexes of ruthenium(II) 18 have similar drawbacks, and due to their shorter lifetimes, timeresolved measurement of these probes is not possible with existing time-resolved fluorescence plate readers.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence-Based Cell Viability Screening Assays Using Water-Soluble Oxygen Probes

Hynes

Floyd

Soini³

et al. 2003

SLAS Discovery

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A simple luminescence-based assay for screening the viability of mammalian cells is described, based on the monitoring of cell respiration by means of a phosphorescent water-soluble oxygen probe that responds to changes in the concentration of dissolved oxygen by changing its emission intensity and lifetime. The probe was added at low concentrations (0.3 microM to 0.5 nM) to each sample containing a culture of cells in the wells of a standard 96-well plate. Analysis of oxygen consumption was initiated by applying a layer of mineral oil on top of each sample followed by monitoring of the phosphorescent signal on a prompt or time-resolved fluorescence plate reader. Rates of oxygen uptake could be determined on the basis of kinetic changes of the phosphorescence (initial slopes) and correlated with cell numbers (10(5) to 10(7) cells/mL for FL5.12 lymphoblastic cell line), cell viability, or drug/effector action using appropriate control samples. The assay is cell noninvasive, more simple, robust, and cost-effective than existing microplate-based cell viability assays; is compatible with existing instrumentation; and allows for high-throughput analysis of cell viability.

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A Water-Soluble Luminescence Oxygen Sensor

Cited by 84 publications

References 20 publications

Fast responsive, optical trace level ammonia sensor for environmental monitoring

Fast responsive, optical trace level ammonia sensor for environmental monitoring

Uncoupling of protein‐3 induces an uncontrolled uncoupling of mitochondria after expression in muscle derived L6 cells

Fluorescence-Based Cell Viability Screening Assays Using Water-Soluble Oxygen Probes

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