TY - JOUR
T1 - Analysis of intracellular oxygen and metabolic responses of mammalian cells by time-resolved fluorometry
AU - O'Riordan, Tomás C.
AU - Zhdanov, Alexander V.
AU - Ponomarev, Gelii V.
AU - Papkovsky, Dmitri B.
PY - 2007/12/15
Y1 - 2007/12/15
N2 - A simple, minimally invasive methodology for the analysis of intracellular oxygen in populations of live mammalian cells is described. Loading of the cells with the phosphorescent O2-sensing probe, MitoXpress, is achieved by passive liposomal transfer or facilitated endocytosis, followed by monitoring in standard microwell plates on a time-resolved fluorescent reader. Phosphorescence life-time measurements provide accurate, real-time, quantitative assessment of average oxygen levels in resting cells and their alterations in response to stimulation. Analytical performance of the method is examined, optimized, and then demonstrated with different suspension and adherent cell lines including Jurkat, PC12, A549, HeLa, SH-SY5Y, and C2C12, by monitoring responses to mitochondrial uncouplers, inhibitors, plasma membrane depolarization, and Ca2+ effectors. The assay provides relevant, information-rich data on cellular function and metabolism. It allows monitoring of small, rapid, and transient changes in cell respiration and screening of new chemical entities.
AB - A simple, minimally invasive methodology for the analysis of intracellular oxygen in populations of live mammalian cells is described. Loading of the cells with the phosphorescent O2-sensing probe, MitoXpress, is achieved by passive liposomal transfer or facilitated endocytosis, followed by monitoring in standard microwell plates on a time-resolved fluorescent reader. Phosphorescence life-time measurements provide accurate, real-time, quantitative assessment of average oxygen levels in resting cells and their alterations in response to stimulation. Analytical performance of the method is examined, optimized, and then demonstrated with different suspension and adherent cell lines including Jurkat, PC12, A549, HeLa, SH-SY5Y, and C2C12, by monitoring responses to mitochondrial uncouplers, inhibitors, plasma membrane depolarization, and Ca2+ effectors. The assay provides relevant, information-rich data on cellular function and metabolism. It allows monitoring of small, rapid, and transient changes in cell respiration and screening of new chemical entities.
UR - https://www.scopus.com/pages/publications/37249064670
U2 - 10.1021/ac701770b
DO - 10.1021/ac701770b
M3 - Article
C2 - 18001129
AN - SCOPUS:37249064670
SN - 0003-2700
VL - 79
SP - 9414
EP - 9419
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 24
ER -