Talk:Development MitoFit proficiency test
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Revision as of 16:38, 2 October 2018 by Laner Verena (talk | contribs)
Guidelines for the MitoFit Proficiency Test
Registration sheet: #### (PDF zum Herunterladen)?
Please indicate if you are a beginner or an advanced O2k user (guidelines for self-assessment: ####)?
Required equipment:
- Oroboros O2k, Series A-H (2004–2018)
- DatLab 7.3 (provided)
- PC/Laptop, Windows XP+
- O2k-Titration-Set with Hamilton syringes
Experiments are to be performed individually (one O2k instrument per participant), without any further help. Manuals and SOPs can be used.
Material:
- HEK cells, titrated with the “Sample” Hamilton syringe
- Respiration medium: MiR05-Kit (provided)?
- SUIT 1 and 2 pce (D## and D##)?
Settings:
- Set slope smoothing to 20 data points at the beginning the measurement
- Use DL-Protocol for air calibration.
- Use low O2 warning function and set it to 30 µM.
- Add MiR05-Kit lot number, preparation date, cell lot and preparation date in the Experiment/Sample/Comments field
- Theoretical cell count: 1 million cells/mL
- Routine respiration: wait until steady-state, but max 15 min
- RP1: ADP titration (2*10 uL)
- Reoxygenate before uncoupler titration
Data evaluation
- Make a copy of your aircal and RP1-RP2 files into the folder "PT Evaluated".
- Make a copy of the two Excel files \SRV1\Lab\3C_Projects_Data\2017-All Data\2017_Proficiency_test_DSL (Preprint) for evaluation in your personal "PT Evaluated" folder.
- Check the measurement files if all settings are correct, events are set correctly, etc.
- Set marks and copy data to clipboard.
- Use the Excel files for data evaluation. Do not forget to set the proper Sample number and Subsample number you used on tabs "Protocol", "Systematic" and "FCR".
Upon execution of the measurement and data analysis by the participants, DatLab 7.3 (DLD) files with the accompanying MS Excel templates for analysis (provided with the SUIT package) should be submitted to the following website: #####
The measurements are to be anonymously analyzed by the mitochondrial physiology experts at Oroboros Instruments.
