O2k-Workshops: General
General contents of O2k-Workshops: Overview on High-Resolution Respirometry
Introduction: Mitochondrial and cellular respiratory physiology; new challenges for high instrumental performance.
High-resolution respirometry; what makes the difference? Presentation of the OROBOROS Oxygraph-2k
- Optimum system design - the OROBOROS Oxygraph-2k.
- DatLab: on-line recording of oxygen concentration and flux; linear slope versus oxygen flux as a function of time.
- DatLab: the specialized software for high-resolution respirometry; high resolution calibrations.
- Low oxygen and measurement of cellular oxygen consumption – pushing the limits of detection.
O2k and TIP2k: On-line instrumental performance
- Instrumental background: measurement and correction as a function of pO2.
- High-resolution of flux at various steady-states.
- O2k-MultiSensor applications: TPP+, pH, NO
- The Titration-Injection microPump TIP2k: automatic titrations.
- Conceptual and methodological advantages of measurement at physiological low levels of oxygen.
Polarographic oxygen sensor (POS) and O2k service
- Cleaning of anode and cathode.
- Electrolyte and membrane application.
- O2k and TIP2k: instrumental maintenance.
Formats
O2k-Workshop: HRR and O2k-Fluorometry
The O2k-Workshop on HRR and Fluorometry provides a detailed introduction to the OROBOROS Oxygraph-2k (O2k-Core), overview presentations on high-resolution respirometry (HRR), and applications in mitochondrial physiology. An introduction to O2k-Fluorometry is integrated into the programme.
The O2k-Workshop programme is complemented by lectures on mitochondrial physiologoy (general topics; substrate-uncoupler-inhibitor titration SUIT protocols). Short presentations of 'Hot topics in Mitochondrial Physiology' provide participants with the opportunity to present and discuss their work, including projects in progress, methodological details and conceptual aspects
O2k-Workshop: O2k-Fluorometry
The O2k-Fluorometry Workshop is integrated into the general O2k-Workshop, introducing fluorometry combined with high-resolution respirometry (HRR). Following a short introduction to HRR, the O2k-Fluorescence LED2-Module is presented in overview lectures and hands-on sessions. Discussions cover protocols for the measurement of hydrogen peroxide production (Amplex red), mt-membrane potential (Safranin), ATP production (Mg green), and Ca2+ (Ca green).
Following our successful IOC tradition, participants are invited to submit abstracts for short presentations of 'Hot topics in Mitochondrial Physiology'.
Information on O2k-Fluorometry, with resolution of hydrogenperoxide in the nanomolar range: O2k-Fluorescence LED2-Module.
Moreover, O2k-Workshops with a special focus on O2k-Fluorometry are offered (e.g. IOC95). Further O2k-MultiSensor applications are covered in these specific O2k-Workshops
- Measurement of Ca2+, calibration and calculation of free Ca2+ in buffered solutions (O2k-TPP+ and Ca2+ ISE-Module)
- Acidification rate (pH)
- Nitric Oxide (NO)
O2k-Workshop: TPP/ ISE
The O2k-Workshop TPP/ISE is customized for participants interested in O2k-MultiSensor applications, particularly ion selective electrodes (ISE) for measurement of mitochondrial membrane potential (TPP+, TPMP+) and Ca2+. It covers an introduction to basic concepts on mitochondrial membrane potential: proton pumps, ion gradients, electrochemical potential (proton motive force) and the measurement of mt-membrane potential in biological samples (homogenate, permeabilized fibres). Furthermore, data analysis (from TPP+ to mt-membrane potential) constitutes a major topic.
According to the interest of participants, other O2k-MultiSensor applications can be discussed and demonstrated:
- Measurement of Ca2+, calibration and calculation of free Ca2+ in buffered solutions (O2k-TPP+ and Ca2+ ISE-Module)
- Acidification rate (pH)
- O2k-Fluorescence
- Nitric Oxide (NO)
