IOC recommended reading
From Bioblast
General
»Worksheet O2k-Basic: This worksheet contains questions on the O2k and HRR addressed to participants of O2k-Basic Workshops. Answers to these questions are elaborated during the O2k-Workshop.
SUIT protocols for high-resolution respirometry
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- Gnaiger E (2012) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 3rd ed. Mitochondr Physiol Network 17.18. OROBOROS MiPNet Publications, Innsbruck: 64 pp. »Open Access
- The blue book: OXPHOS analysis; CI linked, CII linked, CI+II linked pathways, respiratory states, abbreviations
- Pesta D, Gnaiger E (2012) High-resolution respirometry. OXPHOS protocols for human cells and permeabilized fibres from small biopisies of human muscle. Methods Mol Biol 810: 25-58. »Open Access
- Introduction to high-resolution respirometry (HRR), SUIT protocols and flux control ratios.
- Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial Dysfunction in Drug-Induced Toxicity (Dykens JA, Will Y, eds) John Wiley: 327-352. »Open Access
- Methodological introduction to HRR, with focus on Polarographic oxygen sensor and traditional oxygraphy
- Dramatically different: High-resolution respirometry - the Oxygraph-2k
- Calibration of polarographic oxygen sensors and oxygen concentration in respiration media at air saturation
- From Oxygraph slopes to respiratory flux corrected for background effects
- Phosphorylation or coupling control protocol (CCP) with intact cells
- Titration steps of the CCP
- Experimental example
- Flux control ratios from the CCP
- Intact cells, permeabilized cells and tissue, or isolated mitochondria?
HRR and O2k-Fluorometry
- »Manual: O2k-Fluorescence LED2-Module
- Eigentler A, Fontana-Ayoub M, Gnaiger E (2013) O2k-Fluorometry: HRR and H2O2 production in mouse cardiac tissue homogenate. Mitochondr Physiol Network 18.05(01): 1-6. »Open Access
- »O2k-Fluorometry Publications
TPP/ISE
- O2k-MultiSensor System with Ion Selective Electrodes (ISE) MiPNet15.03
- Determination of membrane potential using TPP+ and an ion selective electrode (ISE) MiPNet14.05
- Rottenberg H (1984) Membrane potential and surface potential in mitochondria: uptake and binding of lipophilic cations. J Membr Biol 81: 127-138.
Further reading
- Gnaiger E (2009) Capacity of oxidative phosphorylation in human skeletal muscle. New perspectives of mitochondrial physiology. Int J Biochem Cell Biol 41: 1837-1845. »Abstract
- Respirometry with permeabilized fibres and isolated mitochondria
- Convergent CI+II electron input and OXPHOS capacity
- Tissue-OXPHOS capacity in human permeabilized muscle fibres and isolated mitochondria
- Tissue-OXPHOS capacity and functional diversity
- Gnaiger E (2001) Bioenergetics at low oxygen: dependence of respiration and phosphorylation on oxygen and adenosine diphosphate supply. Respir Physiol 128: 277-297. – A detailed introduction into high-resolution respirometry with particular emphasis on kinetics and measurements at low oxygen. »Abstract
- Mitochondrial kinetics measured by high-resolution respirometry
- Calibrations and corrections for response time and instrumental background
- Steady-state injection respirometry
- Mitochondrial respiratory control at low oxygen
- Apparent oxygen affinity and catalytic efficiency of mitochondrial respiration
- Effect of ADP and oxygen limitation on ADP/O2 flux ratios
- The low-oxygen environment of the cell: Mitochondria between hypoxic and oxidative stress
- Scandurra FM, Gnaiger E (2010) Cell respiration under hypoxia: Facts and artefacts in mitochondrial oxygen kinetics. Adv Exp Med Biol 662: 7-25. »Open Access
- Isolated mitochondria and intact cells
- Nanomolar resolution of oxygen concentration
- Effects of cell density and activity
- Closed versus open systems
- p50 in vivo
- Gnaiger E, Kuznetsov AV, Schneeberger S, Seiler R, Brandacher G, Steurer W, Margreiter R (2000) Mitochondria in the cold. In: Life in the Cold (Heldmaier G, Klingenspor M, eds) Springer, Heidelberg, Berlin, New York: 431-442. »Abstract
- Isolated mitochondria and permeabilized muscle fibers, MiR05
- Optimization of mitochondrial cold storage • Mitochondrial respiration medium, MiR05 • Mitochondrial cold ischemia-reperfusion injury
- Renner K, Amberger A, Konwalinka G, Kofler R, Gnaiger E (2003) Changes of mitochondrial respiration, mitochondrial content and cell size after induction of apoptosis in leukemia cells. Biochim Biophys Acta 1642: 115-123. »Abstract
- Intact cells
- Cytochrome c oxidase
- Cytochrome c test
- Respiration per million cells, per citrate synthase, per mg protein, or per cytochrome c oxidase activity.