Peer-Reviewed Journal Details
Mandatory Fields
Fercher, A,Ponomarev, GV,Yashunski, D,Papkovsky, D;
Analytical and Bioanalytical Chemistry
Evaluation of the derivates of phosphorescent Pt-coproporphyrin as intracellular oxygen-sensitive probes
Optional Fields
Cellular oxygen Phosphorescent oxygen-sensitive probes Pt-porphyrins Intracellular probe Cell metabolism and mitochondrial function BIOLOGICAL APPLICATIONS CELLS SENSORS MITOCHONDRIA DELIVERY CALCIUM REAGENT ASSAYS POWER
Several new derivatives of the phosphorescent Pt (II)-coproporphyrin (PtCP) were evaluated with respect to the sensing of intracellular oxygen by phosphorescence quenching. Despite the more favorable molecular charge compared to PtCP, self-loading into mammalian cells was rather inefficient for all the dyes, while cell loading by facilitated transport using transfection reagents produced promising results. The PtCP-NH2 derivative, which gave best loading efficiency and S/N ratio, was investigated in detail including the optimisation of loading conditions, studies of sub-cellular localization, cytotoxicity, oxygen sensitivity and long-term signal stability. Being spectrally similar to the macromolecular MitoXpress(TM) probe currently used in this application, the PtCP-NH2 demonstrated higher loading efficiency and phosphorescent signals, suitability for several problematic cell lines and a slightly increased lifetime scale for the physiological range (0-200 mu M O-2). In physiological experiments with different cell types, mitochondrial uncouplers and inhibitors per-formed on a time-resolved fluorescence plate reader, this probe produced the anticipated profiles of intracellular oxygen concentration and responses to cell stimulation. Therefore, PtCP-NH2 represents a convenient probe for the experiments and applications in which monitoring of cellular oxygen levels is required.
DOI 10.1007/s00216-009-3399-z
Grant Details