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Currently Answered Questions from the »Oroboros Ecosystem Agenda«

Aug 2021

  • Aug-05 Th
O2k-Open SupportInfo: improve What is the advantage of using flux control ratios (FCRs)?

FCRs provide information on coupling and pathway control of mitochondrial respiration independent of (i) mt-content in cells or tissues, (ii) purification of isolated mitochondrial preparations, and (iii) determination of tissue mass or mt-markers.
- »Flux control ratio«
- communicated by »Cardoso Luiza« and »Cecatto Cristiane«

Jul 2021

  • Jul-29 Th
O2k-Open SupportInfo: improve Do you know how to export a user-specific DL-protocol (*.DLPU)?

- »Export_DL-Protocol_User_(*.DLPU)« and »O2k-Videosupport:Export DL-Protocol User«
- communicated by »Schmitt Sabine«, »Cecatto Cristiane«, and »Komlodi Timea«

  • Jul-28 We
O2k-Open SupportInfo: improve Need to tailor a SUIT protocol to answer your research question?

See how to modify existing DL-protocols.
- »Enable_DL-Protocol_editing«
- communicated by »Schmitt Sabine«, »Cecatto Cristiane«, and »Komlodi Timea«

  • Jul-21 We
MitoPedia: SUIT Not sure which SUIT protocol to apply?

The SUITbrowser is the perfect tool to find the best SUIT protocol for your research question.
- » SUITbrowser« and »O2k-Videosupport: How to find a DL-Protocol«

- communicated by »Schmitt Sabine« and »Komlodi Timea«

Jun 2021

  • Jun-28 Mo
O2k-Open SupportInfo: improve Do I need to calibrate the fluorescence signal with safranin?

It is possible to run the experiment without calibration, presenting only the fluorescence intensity plot. However, a simple two-point or multiple-point calibration for safranin concentration is possible.
- »Safranin calibration«

  • Jun-16 We
O2k-Open SupportInfo: improve Simultaneous fluctuations in O2 slope neg. and the signal of the Peltier power?

This can indicate an electrical disturbance and changing the electrical supply may resolve this problem.
- »Troubleshooting«
- communicated by »Cecatto Cristiane« and »Schmitt Sabine«

  • Jun-07 Mo
O2k-Open SupportWarning: loss of data Do you observe high fluorescence signal even before adding the dye or sample?

Filters that have shifted from the correct position in the Fluo-Sensors might give rise to higher fluorescence signal than usual.
- »Troubleshooting: fluorescence module« 
- communicated by »Komlodi Timea« and »Schmitt Sabine«

  • Jun-03 Th
O2k-Open SupportWarning: loss of data How to ensure quality control when exchanging the 2-mL and the 0.5-mL chambers

Every time you exchange the chambers, a volume calibration and instrumental O2 background test should be performed.
- »MiPNet22.11 Section 5.4. O2k-chamber volume calibration« and »Oxygen flux - instrumental background«
- communicated by »Komlodi Timea« and »Cecatto Cristiane«

May 2021

  • May-25 Tu
O2k-Open SupportTake care Do you enter the correct instrumental background correction in DatLab for the experimental O2 concentrations?

After performing the instrumental background test at the appropriate O2 range, load the parameters in DatLab to ensure accurate correction for external fluxes.
- »O2 background correction in DatLab

- communicated by »Komlodi Timea« and »Cecatto Cristiane«

  • May-17 Mo
O2k-Open SupportInfo: improve Planning to measure proton flux or mitochondrial membrane potential with ion-selective electrodes in the O2k?

Learn how to assemble the reference electrodes:
- »O2k-Videosupport: Assembly of the reference electrode«
- communicated by »Cardoso Luiza« and »Schmitt Sabine«

  • May-04 Tu
O2k-Open SupportInfo: improve Do you know how to apply the air and zero calibration values in DatLab?

This important quality control step ensures accurate measurement of respiration.
For details on how to apply air and zero calibration in DatLab, see:
»Oxygen calibration - DatLab« and »Troubleshooting« 
- communicated by »Komlodi Timea« and »Cecatto Cristiane«

Apr 2021

  • Apr-27 Tu
O2k-Open SupportTake care Why is the fluorescence signal constantly at 10 V or at 0 V?

Follow these steps to localize and solve the problem:
- »Fluorescence signal is constantly at 10 V« and »0 V fluorescence signal«

  • Apr-20 Tu
O2k-Open SupportWarning: loss of data Noisy fluorescence signal?

Follow these steps to localize and solve the problem:
- »Noisy fluorescence signal«

  • Apr-13 Tu
O2k-Open SupportWarning: loss of data Do you know what to do when the fluorescence signal starts at a negative value?

A negative signal may be caused by wet Fluo-Sensors. For further information:
- »Negative fluorescence signal«

  • Apr-06 Tu
O2k-Open Support Warning: loss of data A damaged OroboPOS-Holder may cause medium to leak from the O2k-Chamber.

See how you can check that the OroboPOS-Holders are intact.
- »O2k-OroboPOS-Holder«

Mar 2021

  • Mar-25 Th
O2k-Open SupportInfo: improve O2k-checklist: Get started with an O2k-experiment

The O2k-checklist provides an overview of all essential steps for starting an O2k-experiment.
- » MiPNet20.04_O2k-checklist« 
- » communicated by »Cecatto Cristiane« and »Schmitt Sabine«

Feb 2021

  • Feb-17 We
O2k-Open SupportInfo: improve Do you know how to prepare permeabilized muscle fibers?

The application of high-resolution respirometry in permeabilized muscle fibers provides a sensitive diagnostic tool for the analysis of mitochondrial (dys)function in small biopsy samples.
- »O2k-Videosupport: Fiber permeabilization« 
- »Doerrier 2018 Methods Mol Biol« 
- communicated by »Doerrier Carolina« and »Tindle-Solomon Lisa«

  • Feb-08 Mo
O2k-Open SupportInfo: improve Do you know how to properly insert the ISS-tip into the O2k-chamber?

With the stirrer bar still rotating, insert the ISS-tip to the bottom of the O2k-Chamber, so that it points away from you.
- »MiPNet19.03 O2k-cleaning and ISS: Section 2« 
- communicated by »Doerrier Carolina«, »Cecatto Cristiane« and »Komlodi Timea«

  • Feb-02 Tu
O2k-Open SupportInfo: improve DatLab error messages: What do they mean and how to solve them?

- » DatLab error messages« 
- communicated by »Garcia-Souza Luiz F« and »Tindle-Solomon Lisa«

Jan 2021

  • Jan-11 Mo
O2k-Open SupportInfo: improve Are you a beginner or need a refresher on using SUIT protocols with DatLab7.4?

- » O2k-Videosupport: How to perform an experiment with a SUIT DL-Protocol« and » Run DL-Protocol/Set O2 limit in DatLab « 
- communicated by »Komlodi Timea« and Schmitt Sabine«