IOC65

O2k-Workshop on High-Resolution Respirometry: O2k-Basic and TPP-Basic

Schröcken, Vorarlberg, Austria 10 - 15 December, 2011.

Hotel Körbersee

More information on IOC62: http://www.oroboros.at/index.php?nextcourse

For some pictures, see IOC61

Contact: [email protected]

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Combined Workshop O2k-Basic and TPP-Basic with Parallel Sessions

• Groups will be working in parallel sessions (O2k-Basic and TPP-Basic) to adjust the programme optimally to the specific needs of the participants in different workpackages.

Preliminary Programme

A: Introductory lectures and hands-on experiments for basic HRR in small working teams:

A1. General HRR topics

• O2k-assembly
• Polarographic oxygen sensor (OroboPOS) service
• POS calibration
• DatLab software: principles and operation
• Titration-Injection-microPump - TIP2k.

A2. Hands-on experiments with the O2k

• Measurement of instrumental background oxygen flux, manual and TIP2k-feedback control
• Intact cells: Respirometric reference assay with yeast, phosphorylation control protocol
• Preparation of permeabilized muscle fibres
• HRR with permeabilized muscle fibres.

A3. Presentation and interpretation of oxygen flux

• Normalization of respiration per tissue or per mitochondrial marker
• Coupling control ratios
• Substrate control ratios
• Generalized flux control ratios.

B: Introductory lectures and hands-on experiments for basic TPP⁺ (mitochondrial membrane potential) in small working teams

• Hands-on MultiSensor TPP⁺ applications, see TPP special interest group during an International Oxygraph Course
• From TPP⁺ to mitochondrial membrane potential: unspecific binding, partitioning, chemical side effects.

C: General topics

• Overview on the O2k and DatLab: High-resolution respirometry (HRR)
• Overview on O2k-MultiSensor applications

• Mitochondrial membrane potential (TPP+ electrode)
• Acidification (pH electrode), pH-stat feedback control
• Nitric oxide (amperometric)

• Introduction to fundamental elements of mitochondrial respiratory physiology
• Protocols in HRR: Design and examples of experimental protocols with intact cells, permeabilized cells and tissues, isolated mitochondria.
• Quality control - towards quality assurance

D: Special interest groups

• Oxygen kinetics: experimental examples with TIP2k-feedback control (oxystat) and data analysis,
• ADP kinetics: experimental examples with TIP2-steady state control and data analysis,
• Advanced experimental protocols

Tutors

• Erich Gnaiger (Innsbruck Medical University, AT; OROBOROS INSTRUMENTS, AT)
• Mario Fasching (OROBOROS INSTRUMENTS, AT)
• Dominik Pesta (Innsbruck Medical University, AT)
• Zuzana Sumbalova (OROBOROS INSTRUMENTS, AT)
• Anita Wiethuechter (OROBOROS INSTRUMENTS, AT)