Novel fluorescent oxygen indicator for intracellular oxygen measurements

Ji, Jin; Rosenzweig, Nitsa; Jones, I. C.; Rosenzweig, Zeev

doi:10.1117/1.1483082

Cited by 45 publications

(37 citation statements)

References 24 publications

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“…22 However, there are number of factors that make these derivative unsuitable as an oxygen sensitive dye. For example, emission overlap with autofluorescence, poor chemical-and photostability, 23 limited sensitivity in the physiological range 24 and quenching caused by reactive oxygen species. 25 Platinum and palladium based probes have phosphorescent decays allowing encapsulation which protects it from the applied environment.…”

Section: Introductionmentioning

confidence: 99%

Single photon counting fluorescence lifetime detection of pericellular oxygen concentrations

2012

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Abstract. Fluorescence lifetime imaging microscopy offers a non-invasive method for quantifying local oxygen concentrations. However, existing methods are either invasive, require custom-made systems, or show limited spatial resolution. Therefore, these methods are unsuitable for investigation of pericellular oxygen concentrations. This study describes an adaptation of commercially available equipment which has been optimized for quantitative extracellular oxygen detection with high lifetime accuracy and spatial resolution while avoiding systematic photon pile-up. The oxygen sensitive fluorescent dye, tris(2,2'-bipyridyl)ruthenium(II) chloride hexahydrate ½RuðbipyÞ 3 2þ , was excited using a two-photon excitation laser. Lifetime was measured using a Becker & Hickl time-correlated single photon counting, which will be referred to as a TCSPC card. ½RuðbipyÞ 3 2þ characterization studies quantified the influences of temperature, pH, cellular culture media and oxygen on the fluorescence lifetime measurements. This provided a precisely calibrated and accurate system for quantification of pericellular oxygen concentration based on measured lifetimes. Using this technique, quantification of oxygen concentrations around isolated viable chondrocytes, seeded in three-dimensional agarose gel, revealed a subpopulation of cells that exhibited significant spatial oxygen gradients such that oxygen concentration reduced with increasing proximity to the cell. This technique provides a powerful tool for quantifying spatial oxygen gradients within three-dimensional cellular models.

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Section: Introductionmentioning

confidence: 99%

Single photon counting fluorescence lifetime detection of pericellular oxygen concentrations

2012

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“…Furthermore, luminescence imaging provides high spatial resolution such that exquisite images of oxygen dynamics at the level of a single cell can be acquired. For example, a number of techniques have been developed for intracellular oxygen measurements that involve loading cells (a) with luminophores such as pyrene butyl rhodamine esters (43), ruthenium complexes with a-diimine ligands (44)(45)(46)(47)(48), and various metalloporphyrins (49)(50)(51)(52), or (b) with nanoparticles such as dye-encapsulated liposomes (53) and lipobeads (54), and probes encapsulated by biologically localized embedding (PEBBLE) sensors (55)(56)(57). Thin-film sensing layers comprising a luminophore immobilized in a gas-permeable matrix have also been used for imaging oxygen fluxes.…”

Section: Methods Of Oxygen Detectionmentioning

confidence: 99%

Combined Imaging and Chemical Sensing with a Disposable Microscope Objective Chemical Sensor

Hartzell¹,

Danowski²,

Pantano³

2007

Applied Spectroscopy Reviews

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This manuscript provides a concise review of the combined imaging and chemical sensing (CICS) technique performed with imaging fiber chemical sensors (IFCSs) and represents the first demonstration of CICS using a disposable microscope objective chemical sensor (dMOCS). The dMOCS assembly comprises two pieces that clamp around an imaging system's objective and a third adjoining piece that positions a sensing layer onto a sample's surface and within the objective's field of view. The prototype layer was oxygen-sensitive and was fabricated using a luminescent ruthenium metal complex and a gas permeable polymeric membrane. O 2 -sensitive dMOCSs were characterized with respect to their imaging capabilities, sensitivity, and temporal response, and the feasibility of performing a high-content screening assay was demonstrated with preliminary measurements of oxygen dynamics from living cells. While IFCSs are a requisite for analyzing remote samples that cannot be brought to a microscope stage, for those samples that can, CICS with a dMOCS possesses advantages with respect to spatial resolution, ease of fabrication, cost, and viewing area.

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“…For this reason, oxygen has gained a great deal of interest in the research community for cell-level detection. Single-cell resolution of oxygen concentration levels has been demonstrated using ruthenium chloride complex in J774 murine macrophages (0-280-M concentration range) [25], a more stable ruthenium diimine complex [56], a ruthenium complex integrated onto a fiber-optic probe at 20-250 M with 25-M resolution [45], and a ruthenium diimine complex on the end of a submicron fiber-optic probe (15.6-625 M) [57]. A combination of ruthenium complex and platinum porphyrins has been used to measure both oxygen and pH using fluorescence lifetime rather than intensity, in order to circumvent the instability effects often associated with ruthenium [58].…”

Section: A Luminescencementioning

confidence: 99%

Sensor Technologies for Monitoring Metabolic Activity in Single Cells—Part I: Optical Methods

2004

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A review of optical, chemical, and biological sensors to detect metabolic activity at the single-cell level is presented in the context of the development of lab-on-a-chip research instrumentation. The sensors reviewed include optical sensors, at both research and commercial levels, that can optically detect intracellular metabolites including adenosine triphosphate, nicotinamide-adenine dinucleotide, reduced flavin adenine dinucleotide, and other metabolites, including oxygen, carbon dioxide, and glucose. Methods to optically detect pH changes which are a general indicator of activity in extracellular space are also briefly reviewed. Performance metrics such as sensitivity, sensor size, drift, time response, and sensing range are included when available. Highly suitable optical sensor technologies for monitoring cellular metabolic activity include luminescent (fluorescent, phosphorescent, and chemiluminescent) and colorimetric optical probes. Different approaches to extracting luminescent and colorimetric information are reviewed, including benchtop techniques, fiber-optic approaches, and the use of probes encapsulated by biologically localized embedding. A brief discussion of alternate optical sensor technologies, such as surface plasmon resonance and infrared absorption spectroscopy, is also presented.

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Novel fluorescent oxygen indicator for intracellular oxygen measurements

Cited by 45 publications

References 24 publications

Single photon counting fluorescence lifetime detection of pericellular oxygen concentrations

Single photon counting fluorescence lifetime detection of pericellular oxygen concentrations

Combined Imaging and Chemical Sensing with a Disposable Microscope Objective Chemical Sensor

Sensor Technologies for Monitoring Metabolic Activity in Single Cells—Part I: Optical Methods

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