Abstract
Cell and tissue oxygenation in vivo is maintained within narrow physiological range and its significant deviations from the norm can lead to pathological states. Real‐time monitoring of this parameter and oxygen consumption rate (OCR) in respiring samples provides valuable tool for cell metabolic assessment. We describe two new phosphorescent probes for the analysis of intracellular and extracellular O2 (icO2, ecO2) compatible with multiple detection platforms (ratiometric‐intensity and phosphorescence lifetime based) which are based on the combination of an oxygen‐sensitive PtTFPP dye and PFO antennae dye embedded in cationic or anionic polymer nanoparticles. The icO2 probe MM2 delivered in mammalian cells by passive means was applied in high‐resolution imaging experiments and monitoring of oxygenation within monolayer of adherent fibroblast cells and 3D neuronal spheroids by μs‐FLIM (PLIM), confocal microscopy and microplate reader analysis. The ecO2 probe MM2X was evaluated in OCR measurements in 96‐well microplates and perfusion microchambers. These probes can be used in drug toxicity, cell energy budget and cell physiology studies where high‐resolution 3D O2 imaging and high‐throughput analysis are required. Supported by the Research Executive Agency (REA) of the EU PITN‐GA‐2010–264772 (ITN CHEBANA) and SFI 07/IN.1/B1804 grant.
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