Luxcel Biosciences Limited

University College Cork

Mocon, Inc.

Agilent Technologies, Inc.

Dmitri Boris Papkovsky

Richard Fernandes

Ingo Klimant

Andreas Fercher

Sergey Borisov

Alexander Zhdanov

Daniel W Mayer

John Charles Alderman

James Nial Hynes

Rosemary O'Connor

A method of determining oxygen concentration, metabolic activity, and/or the effects of a test substance on the metabolic activity of a live cell sample by photoluminescence quenching technique employing a photoluminescent probe that self-loads sans any loading reagent into the cells of the cell sample. The probe comprises a plurality of polymeric particles each comprising an amphiphilic cationic polymer matrix having a hydrophobic core and a hydrophilic positively charged surface provided by quaternary amino groups, and a hydrophobic oxygen-sensitive photoluminescent dye embedded in the hydrophobic core.

Method and probe for monitoring oxygen status in live mammalian cells

The invention is based on the use of photoluminescent probes for intracellular sensing of oxygen, especially assaying intracellular oxygen concentration. The photoluminescent probe comprises a suspension of polymeric particles having an average diameter in the 20 nm to 100 nm range, formed from an amphiphilic cationic co-polymer which is oriented in the formed particle to provide a hydrophobic core and a hydrophilic shell. The probe includes a hydrophobic oxygen-sensitive photoluminescent dye such as Pt-tetrakis(pentafluorophenyl)porphine, PtPFPP embedded in the hydrophobic core of the particle, and the co-polymer includes quaternary ammonium groups which provide hydrophilic and cationic character to the particle shell. The photoluminescent probe, which in use is provided in the form of an aqueous suspension of probe, is incubated with live mammalian cells in a suitable growth medium for a period of time to allow the probe particles passively load into the cells. Oxygen can then be sensed by detecting a photoluminescent signal of the photoluminescent probe, which can be correlated with oxygen status, for example oxygen concentration or changes in oxygen concentration/levels, using existing techniques.

Flexibly deployable, discrete, target-analyte sensitive particulate probes and methods of manufacturing and using. The probes each comprise a porous scaffold particle coated with an optically-active, target-analyte sensitive material. The scaffold particle has at least one of (i) a volume of 0.5 to 500 mm, and (ii) a largest dimension of 2 to 20 mm.

Individually and flexibly deployable target-analyte sensitive particulate probes and method of making and using

A self-contained, remotely interrogatable, autonomously positionable, pressure probe and methods of manufacturing and using. The probe () includes (i) a hermetically sealed, flexible, gas impermeable sachet () capable of equilibriating to a surrounding pressure, (ii) an optically-active, target-analyte partial pressure sensitive material () within the sachet, and (iii) a gaseous headspace () within the sachet containing a known concentration of the target-analyte (A).

Photoluminescent pressure probe

A method for monitoring consumption or release of a gaseous analyte such as oxygen by a liquid sample under investigation includes providing a cuvette () having an elongate narrow tube () of a material which is substantially gas impermeable and which is at least partly transparent to measurement excitation radiation and emission radiation along some of the length of the tube. The tube () has a cross-sectional area of under 1 mm. The sample () is loaded into the cuvette (), the sample being in contact with a probe in the tube (), the probe being sensitive to the gaseous analyte, and the liquid having at least one surface and an associated headspace (). The cuvette, the sample, and the probe are equilibrated at a target measurement temperature. Excitation radiation is directed at a sampling zone of the tube () and which is distal from the headspace (), while maintaining the cuvette at the measurement temperature. The emitted radiation is measured and analysed to determine consumption or release by the sample of the gaseous analyte.

Assessment of consumption or release of a gaseous analyte from biological or chemical samples

A sensor material of the type comprising a long-decay photoluminescent, protonable dye embedded in a suitable polymeric matrix, is used for generating a specific optical response to two different analytes present in a sample, thus allowing selective determination of the two analytes in the sample. Also described is a method for the simultaneous sensing of a first and second analyte in a sample. The method comprises the steps of irradiating a sensor material of the type comprising a long-decay photoluminescent protonable dye embedded in a suitable polymeric matrix with light of one or two wavelengths, determining photoluminescence intensity and lifetime signals originating from the sensor, and correlating the photoluminescence intensity signal with a concentration of the first analyte and the photoluminescence lifetime signal(s) with the concentration of the second analyte.

Sensor material and uses thereof to simultaneously sense two analytes: oxygen and pH or acidic/basic gaseous ions

A calibration tool for use in combination with a photoluminescent oxygen-sensitive working probe and an analytical instrument capable of reading the working probe. The calibration tool is effective for achieving two-point calibration of the analytical instrument, and includes at least first and second solid state compositions having different sensitivities to oxygen. The first composition is an oxygen-sensitive photoluminescent dye that is the same as that in the working probe, embedded within an oxygen-permeable carrier matrix that is the same as that in the working probe. The second composition is an oxygen-sensitive photoluminescent dye that is the same as that in the first composition, embedded within a carrier matrix that is different from that in the first composition. The oxygen sensitivity of the second composition is less than the oxygen sensitivity of the first composition.

Calibration card for photoluminescent oxygen sensors

An oxygen sensitive probe comprises a monofunctional derivative of an oxygen-sensitive photoluminescent dye covalently attached to a water soluble and/or hydrophilic macromolecular carrier. The probe may be a chemical conjugate of a monofunctional phosphorescent porphyrin dye and a poly(ethyleneglycol), polypeptide or polysaccharide.

Oxygen sensitive probe

An assay device () comprises a base () and glass plate lid (). The base () has an array of shallow microwells (), each having a flat rim (), all rims being co-planar. When the lid () is placed on the base () a thin capillary gap () is formed on each rim, acting as a liquid seal for a microwell chamber. The liquid is excess sample liquid and further excess is accommodated in overspill cavities () between the microwells (). Because of the liquid seal and shallow configuration the benefits of microfluidic devices are achieved together with the handling convenience and use of conventional detection equipment of conventional microplate devices.

Growing community of inventors

Blarney, Ireland

Dmitri Boris Papkovsky