Baglivo 2022b ESCI Bari

From Bioblast
Baglivo Eleonora
Baglivo E, Cardoso LHD, Cecatto C, Gnaiger E (2022) Statistical analysis of instrumental reproducibility in high-resolution respirometry. 56th Annual Scientific Meeting of the European Society for Clinical Investigation.

Link: 56th Annual Scientific Meeting of the European Society for Clinical Investigation

Baglivo E, Cardoso LHD, Cecatto C, Gnaiger E (2022)

Event: ESCI 2022 Bari IT

Evaluation of instrumental reproducibility is an essential component of quality control ΜΆ defined as standard operating procedures ΜΆ to quantify the precision and limit of detection of an analytical procedure.

Instrumental tests implemented as standard operating procedures in high-resolution respirometry are the sensor test and the chamber test. The sensor test includes calibrations of the signal of the polarographic oxygen sensor (POS) in terms of oxygen concentration cO2 [Β΅M] to evaluate the performance of the POS. The chamber test (instrumental O2 background test) focuses on the slope dcO2/dt to determine oxygen consumption by the POS and backdiffusion into the chamber [1]. We evaluated instrumental tests of 48 Oroboros O2k chambers obtained from a 3-year study on MiR05-Kit (Oroboros Instruments), carried out in the absence of sample.

Stability of oxygen calibration signals at air saturation and zero oxygen was monitored up to 8 months. The maximum drift over 1 to 3 days was 0.05 pmolβˆ™sβˆ’1βˆ™mLβˆ’1, with no persistence over time since drift was < 0.004 pmolβˆ™sβˆ’1βˆ™mLβˆ’1 for a time interval of one month, corresponding to a drift per day of 0.2 % of the signal at air saturation. Instrumental O2 background dcO2/dt was stable within Β±1 pmolβˆ™sβˆ’1βˆ™mLβˆ’1 at different O2 concentrations when measured at monthly intervals.

Taken together, these results confirm the instrumental limit of detection of volume-specific O2 flux at Β±1 pmolβˆ™sβˆ’1βˆ™mLβˆ’1. Following the standard operating procedures applied in the present study provides an instrumental proficiency test to ensure the unique reproducibility in high-resolution respirometry.

β€’ Keywords: High-resolution respirometry, Quality control, Proficiency test, Reproducibility, Limit of detection β€’ Bioblast editor: Plangger M β€’ O2k-Network Lab: AT Innsbruck Oroboros, AT Innsbruck Gnaiger E, AT Innsbruck MitoFit


Affiliations

Oroboros Instruments GmbH, Innsbruck, Austria.

References

  1. Gnaiger E (2001) Bioenergetics at low oxygen: dependence of respiration and phosphorylation on oxygen and adenosine diphosphate supply. Respir Physiol 128:277-97.

List of abbreviations, terms and definitions - MitoPedia

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Labels: MiParea: Respiration, Instruments;methods 





HRR: Oxygraph-2k 


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