Talk:O2k-Workshops in O2k-Network Labs
Preparation for a locally organized O2k-Workshop
Organization and programme
1A. Participants
- How many participants are expected?
- Is it a homogenous group of either (a) experienced users or (b) unexperienced interested participants? Or is the group mixed?
- Is a list of participants available, indicating participants who are O2k-Users?
- Are name tags prepared?
1B. O2k-Demo
- For experienced users, a hands-on session on instrumental assembly and service is important, and a corresponding session should be organized in small groups.
- For an unexperienced group, an O2k-Demo oriented workshop is perferrably conducted in a seminar room.
- Is a demo experiment to be prepared with biological samples provided locally (tissue, cultured cells)? Usually we provide freeze-dried yeast as biological sample for a demo experiment of respiration in intact cells (see below). The demo experiment can be run in a seminar room.
- Is a projector available for PowerPoint presentations and for connecting the laptop connected with DatLab to the O2k?
Instruments and accessories
2A. Which and how many OROBOROS Oxygraph-2k are locally available?
- Specify O2k-Series number(s) of demo instrument.
- Are O2k-Users invited to bring their instrument for practice and guided service? Do they bring their laptop for DatLab?
- Are there any spare parts needed from OROBOROS INSTRUMENTS?
2B. Which accessories and consumables are locally available?
- OroboPOS-Service Kit (necessary) including OroboPOS-Membranes and OroboPOS-Electrolyte Powder (or freshly prepared electrolyte).
- ISS-Integrated Suction System (necessary).
- Titration Set (some Hamilton syringes are specifically required for manual titrations).
- HRR-Dissection Set (required if permeabilized tissue preparation is to be demonstrated).
- PBI-Shredder HRR-Set (required if tissue homogenate should be prepared).
- Microbalance-Set (required if tissue mass, particularly wet weight, should be determined).
2C. Which O2k-Modules are locally available?
- TIP2k-Module (optional)
- O2k-Fluorescence LED2-Module (recommended)
- O2k-TPP+ and Ca2+ ISE-Module (only for special ISE workshop)
- O2k-pH ISE-Module (only for special O2k-MultiSensor workshop)
- O2k-NO Amp-Module and separate NO sensor (only for special O2k-MultiSensor workshop)
Chemicals and consumables
3A. OroboPOS service
- Ammonia solution concentrated, a few ml of 25% concentrated ammonia (fresh).
- A hood or safe place (separate room) for ammonia cleaning of the OroboPOS (ask for details).
3B. Demo O2k-Core (cell respiration)
Further details: A mitochondrial reference assay for high-resolution respirometry using freeze-dried Bakerโs yeast. ยปMiPNet15.09. Yeast: HRR Reference Assay.
- Na-P buffer
- 100% Ethanol in wash bottle
- 70% EtOH in wash bottle
- Deionised water in wash bottle
- 1 ml pipette
- Gloves
3C. Demo O2k-Fluorometry (cell respiration and extracellular hydrogen peroxide production)
Further details: O2k-Fluorometry: Amplexยฎ UltraRed using freeze-dried bakerโs yeast. ยปMiPNet18.06 Amplex-Yeast.
In addition to the above, these chemicals are required:
- Amplex red.
- Horseradish peroxidase.
- H2O2 solutions for calibration (see Amplex red).
- Ethanol and glucose stocks for substrate titrations.
- CCCP (uncoupler; alternatively FCCP) dissolved in ethanol or DMSO.
We bring: 20 mg yeast (pre-weighted) and 1 full package of yeast (fresh).