Fasching IOC61-TPP group
|Fasching M, Sumbalova Z (2011) TPP special interest group during IOC61. MiPNet16.01.|
The simultaneous measurement of Mitochondrial membrane potential with the TPP method and respiration under HRR conditions with the Oroboros ISE system is explained, practiced and discussed in a "hands-on" approach.
TPP special interest group during IOC61
During IOC61 Oroboros Instruments offers a TPP special interest group during an International Oxygraph Course covering the utilization of the Oroboros ISE system to measure mitochondrial membrane potential. Some topics of general interest will be covered together with the participants from the "main course". This special interest groups is intended as a workshop, therefore the exact program will depend on the level of experience and interests of the participants. In an introductory session the principals of measuring mitochondrial membrane potential with a TPP-electrode are covered. The different steps of planning and performing a TPP are discussed. In "hands-on" sessions the participants will follow MiPNet15.03_O2k-MultiSensor-ISE to assembly TPP and reference electrodes, set up an oxygraph-2k with these electrodes, record an instrumental oxygen background in their presence, calibrate the electrodes and determine the effect chemicals used in a biological experiment on the electrodes. In a hands-on experiment this set up will be used in the experiment with tissue homogenate from mouse brain for measurement of changes in mitochondrial membrane potential simultaneously with changes in respiration in the course of SUIT protocol. Calculation of mitochondrial membrane potential from measurements with a TPP electrode will be intensively discussed and facilitated by the provided spreadsheet templates. Specificity of several SUIT protocols will be discussed. Interested participants can join a special group combining HRR with measuring Mitochondrial membrane potential of permeabilized fibres.