TITLE

Evaluation of the derivates of phosphorescent Pt-coproporphyrin as intracellular oxygen-sensitive probes

AUTHOR(S)
Fercher, Andreas; Ponomarev, Gelii V.; Yashunski, Dmitri; Papkovsky, Dmitri
PUB. DATE
March 2010
SOURCE
Analytical & Bioanalytical Chemistry;Mar2010, Vol. 396 Issue 5, p1793
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
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™ 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 μM O2). In physiological experiments with different cell types, mitochondrial uncouplers and inhibitors performed 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.
ACCESSION #
48191126

 

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