Talk:Development MitoFit proficiency test

Guidelines for the MitoFit Proficiency Test

Required equipment:

• Oroboros O2k, Series A-H (2004–2018)
• DatLab 7.3 (provided)
• PC/Laptop, Windows XP+
• O2k-Titration_Set with Hamilton microsyringes

Experiments are to be performed:

• individually (one O2k instrument per participant)
• without any further help
• Manuals and SOPs can be used, see: O2k-technical_support_and_open_innovation#O2k-Manual

Beginner or advanced user? - Self assessment:

Please indicate if you are a beginner or an advanced O2k user, filling out the questionnaire:

Send the questionnaire to [[1]]

Ready to participate? Send your registration form to: [[2]]

Important informations

• How much time is needed for the execution of the test?

Level 1. Air calibration and Instrumental background oxygen flux: ca. 3 hours

Level 2. SUIT-1 or SUIT-2: ca. 3 hours, including air calibration

Level 3. Data analysis: 1 hour

Within the timespan of one workday, your personal proficiency is comprehensively evaluated, using your own O2k and your laboratory equipment

• Is there any scientific objective?

Together with a literature review and preliminary data from the Oroboros MitoFit laboratory, we are going to publish the test results in collaboration with the Department for Medical Statistics, Informatics and Health Economics, Medical University of Innsbruck, as part of the K-Regio Project MitoFit.

With the participation in the MitoFit Proficiency Test, you agree on the use of experimental data collected for the test, to be used for research purposes in an anonym way.

• Additional resources:

For a comprehensive list of DatLab menus: http://wiki.oroboros.at/index.php/MitoPedia:_DatLab

MitoPedia – SUIT: http://wiki.oroboros.at/index.php/MitoPedia:_SUIT

Flux analysis: http://wiki.oroboros.at/index.php/MiPNet19.18E_O2_flux_analysis_-_DatLab_6

DLP: List of titrations, marks, overview on the website.

Information to be included/udpated in the MiPNet?

Material:

• HEK cells, titrated with the “Sample” Hamilton syringe
• Respiration medium: 1 vial 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.