Djafarzadeh 2017 J Vis Exp
|Djafarzadeh S, Jakob SM (2017) High-resolution respirometry to assess mitochondrial function in permeabilized and intact cells. J Vis Exp 8.doi: 10.3791/54985..|
Abstract: A high-resolution oxygraph is a device for measuring cellular oxygen consumption in a closed-chamber system with very high resolution and sensitivity in biological samples (intact and permeabilized cells, tissues or isolated mitochondria). The high-resolution oxygraph device is equipped with two chambers and uses polarographic oxygen sensors to measure oxygen concentration and calculate oxygen consumption within each chamber. Oxygen consumption rates are calculated using software and expressed as picomoles per second per number of cells. Each high-resolution oxygraph chamber contains a stopper with injection ports, which makes it ideal for substrate-uncoupler-inhibitor titrations or detergent titration protocols for determining effective and optimum concentrations for plasma membrane permeabilization. The technique can be applied to measure respiration in a wide range of cell types and also provides information on mitochondrial quality and integrity, and maximal mitochondrial respiratory electron transport system capacity.
Labels: MiParea: Respiration, Instruments;methods
Preparation: Isolated mitochondria
Coupling state: OXPHOS, ET Pathway: N, S, CIV HRR: Oxygraph-2k