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Phosphorescent nanoparticle based probes for imaging and measurement of cellular oxygen
Author(s) -
Kondrashina Alina,
Dmitriev Ruslan,
Borisov Sergey,
Klimant Ingo,
Zhdanov Alexander,
Papkovsky Dmitri
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.576.2
Subject(s) - phosphorescence , biophysics , live cell imaging , single cell analysis , chemistry , oxygen , confocal microscopy , intracellular , nanotechnology , materials science , cell , microbiology and biotechnology , optics , fluorescence , biology , biochemistry , physics , organic chemistry
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 O 2 (icO 2 , ecO 2 ) 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 icO 2 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 ecO 2 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 O 2 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.