Practical workshop on the O2k-fluorometer
This event has already taken place.
VU's Institute of Sport, Exercise and Active Living (ISEAL) is hosting a practical scientific workshop on how to use the new O2k fluorometer.
Higher degree by research students and staff in the relevant sport science research areas, including staff and students from other universities, are invited to attend this workshop on high-resolution respirometry (HRR) and O2k fluorometry.
The workshop will be conducted by Professor Erich Gnaiger and Dr Carolina Doerrier from Oroboros Instruments in Austria. It will include a basic introduction to quality control of instrumental performance of the OROBOROS O2k, with integrated on-line analysis and will introduce new features of DatLab 6.
This 2-day workshop will be hosted by Professor David Bishop and Associate Professor Nigel Stepto from the College of Sport & Exercise Science and ISEAL at Victoria University.
Topics
Discussions and experiments in this workshop will include:
- Optimisation of OXPHOS analysis in various mitochondrial (mt) preparations (permeabilised muscle fibres, tissue homogenate, isolated mitochondria)
- Comparison of measurements of mt-membrane potential using safranin (fluorometric) or TPP+ or TPMP+ (potentiometric), and on perspectives of HRR in mitochondrial physiology.
State-of-the-art OXPHOS analysis is extended using mt-preparations. It is used to:
- Evaluate coupling efficiencies and OXPHOS capacities with carbohydrate versus fatty acid substrates
- Diagnose defects in respiratory complexes of the electron transfer system and phosphorylation system.
This workshop will also present the latest developments on substrate-uncoupler-inhibitor titration (SUIT) protocols in HRR using the O2k-fluorescence LED2-module for simultaneous measurement of hydrogen peroxide production (Amplex red®).
This O2k-Workshop is a satellite event to Research to Practice 2016.
Registrations
Registrations close on Friday 9 April.
The registration fee includes:
- attendance at activities over the 2 days of the practical workshop, with 2 presenters from Austria
- lunches, coffee breaks and the Workshop dinner on Tuesday evening.
Registration prices
- Students $350
- Staff /Post Docs $450
To register, please contact Lili Hodnik:
Phone: +613 9919 4054
Email: [email protected].
Program
Day 1 Tuesday 12 April 2016
| Time | Activity |
|---|---|
|
8.45-9.00am |
Registration |
|
9.00-9.15am |
Welcome by Professor David Bishop |
|
9.15-9.30am |
Introduction of participants |
|
9.30-10.30am |
Get started with the O2k |
|
10.30-11.00am |
Coffee break |
|
11.00am-12.15pm |
Evaluation of mt-preparations
|
|
12.15-12.30pm |
Permeabilised fibre preparation - what to take care of? |
|
12.30-1.15pm |
Lunch |
|
1.15-2.00pm |
Phosphorylation protocol for intact cells |
|
2.00-3.00pm |
Comprehensive OXPHOS analysis
|
|
3.00-3.30pm |
Experimental setup 1: OroboPOS - sensor quality control, calibration. |
|
3.30-4.00pm |
Coffee break |
|
4.00-5.00pm |
Experimental setup 2: Calibration of O2k-Fluo Sensors |
|
5.00-5.30pm |
Bioblast wiki and O2k-Network |
|
5.30-6.00pm |
Q&A on HRR and OXPHOS analysis
|
|
6.00pm - |
O2k-Workshop dinner |
Day 2 Wednesday 13 April 2016
| Time | Activity |
|---|---|
|
8.30-10.30am |
Experiment: HRR and O2k-Fluorometry with permeabilised fibres from rat – respiration and extracellular H2O2 production. |
|
10.30-11.00am |
Coffee break |
|
11.00am-12.00pm |
Experiment continued |
|
12.00-12.45pm |
Lunch |
|
12.45-3.30pm |
Data analysis |
|
3.30-4.00pm |
Coffee break |
|
4.00-4.40pm |
Technical support and open innovation |
|
4.40-6.00pm |
Feedback and conclusions
|
When?
This event has already taken place.
