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).

• The workshop programme is complemented by combined lectures ("General topics") and experiments run in "Special interest groups".

• Short presentations of 'Hot topics in Mitochondrial Physiology' extend the discussions on respiratory protocols.

Please register for O2k Basic or TPP Basic according to your specific needs and interest.

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)