Gnaiger 1998 BTK-HRR
|Gnaiger E, Kuznetsov AV, Lassnig B, Fuchs A, Reck M, Renner K, Stadlmann S, Rieger G, Margreiter R (1998) High-resolution respirometry. Optimum permeabilization of the cell membrane by digitonin. In BioThermoKinetics in the Post Genomic Era (Larsson C, Pahlman I-L, Gustafsson L, eds) Chalmers Reproservice, Göteborg:89-95.|
Abstract: Permeabilization of the cell membrane is becoming an established alternative to the isolation of mitochondria in bioenergetic studies of cultured cells and biopsy samples of muscle. Depending on cell type, 5 to 50 million cells are required for a respirometric measurement of endogenous and permeabilized-cell respiration [1,2], using conventional oxygen monitoring instruments. An order of magnitude less cells are required when using high-resolution respirometry  (Tab. 1), which provides an up-to-date standard for general and clinical bioenergetics, including the diagnosis of mitochondrial defects, testing of drugs, oxidative stress and hypoxia-reoxygenation injury [4-6]. We provide an overview of (i) the specific features of high-resolution respirometry, (ii) test experiments for instrumental evaluation, and (iii) digitonin titrations for determining optimum concentrations for cell membrane permeabilization in cultured cells.
Labels: MiParea: Respiration, Instruments;methods
Organism: Human Tissue;cell: Endothelial;epithelial;mesothelial cell, HUVEC Preparation: Intact cells, Permeabilized cells
Regulation: Inhibitor, Oxygen kinetics, Substrate Coupling state: ROUTINE, OXPHOS Pathway: S HRR: Oxygraph-2k, TIP2k