MiPNet16.04 IOC62

                

---

---

MiPNet16.04 IOC62

Innsbruck AT, 2011 Jul 11-13. Oroboros O2k-Workshop on high-resolution respirometry (HRR) and O2k-Fluorometry, IOC62.

Oroboros (2011-07-11) Mitochondr Physiol Network

Abstract:

62th Oroboros O2k-Workshop on high-resolution respirometry and O2k-Fluorometry. Innsbruck, AT; 2011 July 11-13.

• O2k-Network Lab: AT Innsbruck Oroboros

Labels:

HRR: Oxygraph-2k 

ORO, IOC, 2011 

Lecturer

Gnaiger Erich, CEO, Oroboros Instruments

General information

The July-Workshop includes a basic introduction to the Oroboros O2k, overview presentations on high-resolution respirometry (HRR), and applications in mitochondrial physiology. Specific emphasis is placed on hands-on practical sessions with the instrument and the software DatLab 4.3, to introduce the participants into the technical and operational details of the method. An overview is given on the O2k-MultiSensor System, using our TPP+ electrode for simultaneous measurement of mt-membrane potential and oxygen consumption.

As an outlook to INNOVATION, we will present our newest perspectives on the O2k-Fluorometer, which is being developed in the frame of the project MitoCom Tyrol. It is intended not only to develop a new generation instrument, but also to maintain our strategy of options for modular upgrading the current O2k-Models. The workshop programme is complemented by lectures and experiments based on substrate-uncoupler-inhibitor titration (SUIT) protocols. Short presentations of 'Hot topics in Mitochondrial Physiology' extend the discussions on respiratory protocols.

Programme

Monday, 11. July

• 09:00-12:30 Introduction to High-Resolution Respirometry

• 14:00-17:30 Hands-on experiments with the O2k: Calibration and instrumental background oxygen flux

Tuesday, 12. July

• 09:00-12:30 The O2k-Fluorimeter in development: Demo experiment with tissue homogenate

• 14:00-17:30 Hands-on experiments with the O2k: TPP+ calibration and application with tissue homogenate

Wednesday, 13. July

• 09:00-12:30 O2k-MultiSensor High-Resolution Respirometry and data analysis: DatLab and application of Excel files. From TPP+ concentration to mitochondrial membrane potential.

• 14:00-17:30 Mitochondrial respiratory physiology: Protocols with substrate-uncoupler-inhibitor titrations for coupling and substrate control.

Recommended reading

O2k-Core Manual

• New: »O2k-Core Manual.pdf

SUIT protocols for high-resolution respirometry

• Pesta D, Gnaiger E (2012) High-resolution respirometry. OXPHOS protocols for human cells and permeabilized fibres from small biopsies of human muscle. Methods Mol Biol 810:25-58. »Bioblast Access
• 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:327-52. »Bioblast Access

HRR and O2k-Fluorometry

• »Manual: O2k-Fluo 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

Mitochondrial pathways

• 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 - handout to O2k-Workshop participants.

O2k-Workshops are listed as MitoGlobal Events.

• This workshop is a component of the K-Regio project MitoCom.