Baglivo 2022b ESCI Bari

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 ̶ defined as standard operating procedures ̶ 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 [µM] 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 pmol∙s⁻¹∙mL⁻¹, with no persistence over time since drift was < 0.004 pmol∙s⁻¹∙mL⁻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 ±1 pmol∙s⁻¹∙mL⁻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 ±1 pmol∙s⁻¹∙mL⁻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.

• Keywords: High-resolution respirometry, Quality control, Proficiency test, Reproducibility, Limit of detection • Bioblast editor: Plangger M • O2k-Network Lab: AT Innsbruck Oroboros, 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.

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Labels: MiParea: Respiration, Instruments;methods 

HRR: Oxygraph-2k