IOC53 International Workshop on High-Resolution Respirometry
30 July - 4 August 2009, 53rd IOC - International Course on High-Resolution Respirometry. Hotel Tannberg, Schroecken, Vorarlberg, Austria.
The 53rd O2k-Course is the 20th presentation of high-resolution respirometry (HRR) in Schröcken since 1988, for the first time extended to four full days. The O2k-Course includes experiments with biological samples, providing a practical overview of the Oxygraph-2k, with integrated on-line analysis by DatLab 4.3 (new upgrade), applications of the TIP-2k, and perspectives of high-resolution respirometry in mitochondrial physiology. Emphasis is placed on hands-on applications by all participants.
Experienced tutors guide small working groups step-by-step through the approach of HRR. Five Oxygraph-2k (10 chambers) are available for a do-it-yourself application of both hardware and software. Combined with an introduction and demo experiment, it is best to put the O2k into action yourself.
During lunch breaks, sufficient time is available for relaxing walks and talks, to enjoy the refreshing scenery of the alpine environment, or use the spare time for specific tutorials. With DatLab 4.3 we accomplish data analysis on-line during the experiment, providing final results and their graphical presentation by the end of an experimental run. Thus we gain sufficient time to demonstrate MultiSensor perspectives, see the Titration-Injection microPump TIP-2k with new feedback-control in action and practice its simple and automatic operation.
IOC53 Feedback"Thank you very much for this week of course in Austria." - Arnaud Mourier, Stockholm, Sweden
"Thank you for an informative course on use of the Oxygraph-2k and related subjects. It is one of the most interesting courses I have had." - Vinh Than Phung, Norway
"I want to thank you again for a very interesting course both scientifically and social. The PC presentation from Diana was fantastic." - Knut M. Skatvedt, Norway
"Thank you again for the wonderful respirometry course in Schrocken!" - Paul Coen, Pittsburgh, USA
Participants IOC53
- Anthonsen, Stine Roskide University, Roskilde Denmark - sat(at)regionsjaelland.dk
- Asander, Eleonor Lund University, Laboratory for Exp. Brain Research, Lund Sweden - eleonor.asander(at)med.lu.se
- Bartholmei, Diana, OROBOROS INSTRUMENTS, Innsbruck, Austria. - diana.bartholmei(at)oroboros.at (admin)
- Cohen, Paul M. Postdoctoral Associate Division of Endocrinology and Metabolism University of Pittsburgh, Pittsburgh, US - pmc17(at)pitt.edu
- Egelandsdal, Bjorg Prof. Institutt for kjemi, bioteknologi og matvitenskap, Ås, Norway - bjorg.egelandsdal8at)umb.no
- Fasching Mario, PhD, OROBOROS INSTRUMENTS, Innsbruck, Austria. - mario.fasching(at)oroboros.at (Tutor)
- Gnaiger Erich, PhD, Prof., Dept. General and Transplant Surgery, Medical University of Innsbruck, D. Swarovski Research Laboratory Innsbruck, Austria; and OROBOROS INSTRUMENTS; Austria. - erich.gnaiger(at)oroboros.at (Tutor)
- Guelden, Elke Deutschen Diabetes Zentrum, Düsseldorf Germany - elkeguelden(at)yahoo.de
- Mourier, Arnaud Department Laboratory of Medicine, Karolinska Institutet, University Hospital, Stockholm Sweden - arnaud.mourier(at)ibgc.u-bordeaux2.fr
- Phung Vinh - vinh.phung(at)umb.no
- Renner Kathrin, PhD, OROBOROS INSTRUMENTS, Innsbruck, Austria. - kathrin.renner@oroboros.at (Tutor)
- Skatvedt, Knut Manager ILS Laboratories Scandinavia, Oslo, Norway - knut.skatvedt(at)ils-lab.no
- Slinde, Erik - erik(at)imr.no
- Suter Correia Cadena, Sílvia Maria, Prof. Departamento de Bioquímica e Biologia Molecular Universidade Federal do Paraná, Brasil - silvia.cadena(at)ufpr.br
- Szendrödi, Julia Deutschen Diabetes Zentrum, Düsseldorf Germany - julia.szendroedi(at)ddz.uni-duesseldorf.de
- Vaca Cristina, PhD Student, Hospital Universitario de La Princesa, Servicio de Inmunología, Madrid Spain. - cenuvaca(at)yahoo.es
- Wagner, Graham Prof. Department. of Physiology & Pharmacology University of Western Ontario School of Medicine & Dentistry - graham.wagner(at)schulich.uwo.ca
- Zouhar, Petr Institute of Physiology, Academy of Sciences of the Czech Republic Prague Czech Republic - zouhar(at)biomed.cas.cz
Day 1Thursday, 30. July
16:00 Participants arriving in Bregenz: Meeting point at 4:00 pm in Bregenz train station; 1.1 hour drive to Schröcken. Check in at Hotel Tannberg
18:30 Welcome reception Hotel Tannberg
19:00 Dinner
21:00 Erich Gnaiger: Beyond respiratory states 1-2-3-4: electron transport system (ETS), OXPHOS capacity and LEAK respiration - Experimental advances with high-resolution respirometry.
Day 2Friday, 31. July
Principles of high-resolution respirometry - from switching on the Oxygraph2k to the experimental result.
08:30 – 09:30 The O2k system: Introduction and oxygen calibration of the polarographic oxygen sensors (POS).
09:30 – 10:30 Hands-on: Oxygen sensor calibration with DatLab 4.
10:30 Coffee break
11:00 – 12:00 Erich Gnaiger: Experimental protocols for substrate-uncoupler-inhibitor titrations (SUIT protocols): An introduction.
12:00 Lunch break; walk and talk.
15:00 -16:30 Kathrin Renner and Erich Gnaiger: Demo experiment – the Oxygraph-2k and a SUIT protocol.
16:30 Coffee break
17:30 – 18:45 Hands-on: Experiment with the Oxygraph-2k (5 O2k - 10 chambers): SUIT protocol; on-line DatLab analysis.
19:00 Dinner
21:00 Discussion of results and protocol of the demo experiment.
Day 3Saturday, 01. August
08:30 – 09:00 Mario Fasching: Introduction - Instrumental Background
09:00 – 12:00 Hands-on: Instrumental background test with the Oxygraph-2k - Washing and filling the O2k chambers with experimental media; air calibration; instrumental background competition, DatLab background analysis.
A. Instrumental background test for expeirments with cells and isolated mitochondria, from air saturation to zero oxygen concentration, wth automatic TIP-2k titration protocol (see #1.3.B. Instrumental background with dithionite and TIP-2k feedback control. MiPNet 14.6).
B. Instrumental background test for experiments with permeabilized muscle fibers, in the high-oxygen range of 500 to 200 µM. Manual titration of hydrogen peroxide into MiR06 (MiR05 with catalase).
12:00 Lunch break – walk and talk
16:00 – 16:45 Erich Gnaiger: DatLab background analysis – summary.
16:45 Coffee break
Parallel group sessions:
O2k Setup POS Service DatLab Analysis TIP-2k
17:15 – 18:00 Gr. 1 Gr. 2 Gr. 3 Gr. 4
18:00 – 18:45 Gr. 4 Gr. 1 Gr. 2 Gr. 3
19:00 Dinner
21:00 Hot topics: MiPNet Session (10+10 min)
Day 4Sunday, 02. August
Parallel group sessions:
O2k Setup POS Service DatLab Analysis TIP-2k
08:30 – 09:15 Gr. 3 Gr. 4 Gr. 1 Gr. 2
09:15 – 10:00 Gr. 2 Gr. 3 Gr. 4 Gr. 1
10:00 Coffee break
10:30 - 11:30 Mario Fasching: Trouble Shooting
12:00 Walk and welcome at the Alpmuseum uf m Tannberg (www.alpmuseum.at), refreshment in the lake Körebersee or at Hotel Körbersee (we keep the details of timing flexible according to weather conditions).
16:45 Coffee (Hotel Tannberg)
17:15 - 18:45 Mario Fasching: Principles and developments of the O2k-MultiSensorAnalyzer. Demo experiment with a high-resolution TPP electrode.
19:00 Dinner
Day 5Monday, 03. August
09:30 – 10:30 Erich Gnaiger: Respiratory measurements at low oxygen: Aerobic-anoxic transitios and steady-state control
10:30 Coffee break
11:00 - 12:00 Summary of O2k-features and hands-on with DatLab Analysis:
A. The O2k and advanced features of DatLab 4.
B. Practise DatLab 4 with demo data.
12:00 Lunch break – walk and talk
15:00 - 16:00 The O2k system: Open questions
16:00 Coffee
16:30 - 18:00 Erich Gnaiger: Experimental protocols.
a) Phosphorylation control protocol with intact cells: ROUTINE – LEAK – ETS
b) Diagnostic multiple SUIT protocols with mitochondrial preparations.
18:00 - 18:45 Questions and discussion of protocols, analysis and trouble shooting.
19:00 Dinner
21:00 Discussion - Summary – Conclusions
Farewell party of IOC53
Day 6Tuesday, 04. August
Early morning: Departure
