New phosphorescence based probes and techniques for the analysis of cellular oxygen and respiration

Real time monitoring of oxygenation and respiration is on the cutting edge of bioanalysis, including studies of cell metabolism, bioenergetics, mitochondrial function and drug toxicity. This thesis presents the development and evaluation of new luminescent probes and techniques for intracellular O2 sensing and imaging. A new oxygen consumption rate (OCR) platform based on the commercial microfluidic perfusion channel μ-slides compatible with extra- and intracellular O2 sensitive probes, different cell lines and measurement conditions was developed. The design of semi-closed channels allowed cell treatments, multiplexing with other assays and two-fold higher sensitivity to compare with microtiter plate. We compared three common OCR platforms: hermetically sealed quartz cuvettes for absolute OCRs, partially sealed with mineral oil 96-WPs for relative OCRs, and open 96-WPs for local cell oxygenation. Both 96-WP platforms were calibrated against absolute OCR platform with MEF cell line, phosphorescent O2 probe MitoXpress-Intra and time-resolved fluorescence reader. Found correlations allow tracing of cell respiration over time in a high throughput format with the possibility of cell stimulation and of changing measurement conditions. A new multimodal intracellular O2 probe, based on the phosphorescent reporter dye PtTFPP, fluorescent FRET donor and two-photon antennae PFO and cationic nanoparticles RL-100 was described. This probe, called MM2, possesses high brightness, photo- and chemical stability, low toxicity, efficient cell staining and high-resolution intracellular O2 imaging with 2D and 3D cell cultures in intensity, ratiometric and lifetime-based modalities with luminescence readers and FLIM microscopes. Extended range of O2 sensitive probes was designed and studied in order to optimize their spectral characteristics and intracellular targeting, using different NPs materials, delivery vectors, ratiometric pairs and IR dyes. The presented improvements provide useful tool for high sensitive monitoring and imaging of intracellular O2 in different measurement formats with wide range of physiological applications.

Intersections: proceedings of the youth work and music education symposium

The ‘Intersections: Youth Work and Music Education’ Symposium took place on Friday, 27th June 2014 in University College Cork. This event, held in association with the Institute of Social Sciences in the 21st Century (ISS21), was made possible thanks to funding from UCC’s Strategic Research Fund. The principle aim of this seed funding is to generate new research interests and this presented the ideal opportunity for developing collaborative relationships between youth work and music education lecturers, researchers and practitioners across the island of Ireland. This led to the formation of a new ‘Intersections’ research cluster, comprising representatives from four third-level institutions, each of which offers both youth work and music education undergraduate and/or postgraduate programmes, namely: University College Cork; National University of Ireland, Maynooth; Dundalk Institute of Technology; and, University of Ulster. This document presents some preliminary findings from primary research conducted through each of the participating institutions in their local areas. Data was also collected during the symposium, through engaging in small group discussions populated by the event’s participants. The publication and dissemination of this document was included in the original Strategic Research Fund proposal. Its intended audience includes youth workers, music educators, community-based practitioners, policy-makers and academics who are motivated and enthused by the possibilities of music-oriented youth work in Ireland and beyond.