Baglivo 2022 BEC

From Bioblast

Bioblast2022 banner.jpg

Bioenergetics Communications        
Tributes to pioneers in bioenergetics
Gnaiger 2020 BEC MitoPathways
Gnaiger Erich et al โ€• MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1.
        MitoPedia: BEC         MitoPedia: Gentle Science         MitoFit Preprints
Publications in the MiPMap
Baglivo E, Cardoso LHD, Cecatto C, Gnaiger E (2022) Statistical analysis of instrumental reproducibility as internal quality control in high-resolution respirometry. Bioenerg Commun 2022.8.

ยป Bioenerg Commun 2022.08. Open Access pdf
published online 2022-08-04 ยปWatch the presentationยซ

Baglivo Eleonora, Cardoso Luiza HD, Cecatto Cristiane, Gnaiger Erich (2022) Bioenerg Commun

Abstract: BEC.png

Graphical abstract

Evaluation of instrumental reproducibility is a primary component of quality control to quantify the precision and limit of detection of analytical procedures. A pre-analytical instrumental standard operating procedure (SOP) is implemented in high-resolution respirometry consisting of: (1) a daily SOP-POS for air calibration of the polarographic oxygen sensor (POS) in terms of oxygen concentration cO2 [ยตM]. This is part of the sensor test to evaluate POS performance; (2) a monthly SOP-BG starting with the SOP-POS followed by the chamber test quantifying the instrumental O2 background. The chamber test focuses on the slope dcO2/dt [pmolโˆ™sโˆ’1โˆ™mLโˆ’1] to determine O2 consumption by the POS and O2 backdiffusion into the chamber as a function of cO2 in the absence of sample. Finally, zero O2 calibration completes the sensor test.

We applied this SOP in a 3-year study using 48 Oroboros O2k chambers. Stability of air and zero O2 calibration signals was monitored throughout intervals of up to 8 months without sensor service. Maximum drift over 1 to 3 days was 0.06 pmolโˆ™sโˆ’1โˆ™mLโˆ’1, without persistence over time since drift was <0.004 pmolโˆ™sโˆ’1โˆ™mLโˆ’1 for time intervals 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 ยฑ1 pmolโˆ™sโˆ’1โˆ™mLโˆ’1 when measured at monthly intervals. These results confirm the instrumental limit of detection of volume-specific O2 flux at ยฑ1 pmolโˆ™sโˆ’1โˆ™mLโˆ’1. The instrumental SOP applied in the present study contributes to the generally applicable internal quality control management ensuring the unique reproducibility in high-resolution respirometry.
โ€ข Keywords: high-resolution respirometry HRR; polarographic oxygen sensor POS; air calibration; instrumental background; reproducibility; limit of detection; internal quality control IQC; standard operating procedure SOP โ€ข Bioblast editor: Tindle-Solomon L โ€ข O2k-Network Lab: AT Innsbruck Oroboros

Bioenergetics Communications

ORCID:ORCID.png Baglivo Eleonora ORCID.png Cardoso Luiza HD, ORCID.png Cecatto Cristiane, ORCID.png Gnaiger Erich


ยป Baglivo 2022 MitoFit-QC


This work was part of the Oroboros NextGen-O2k project, with funding from the European Unionโ€™s Horizon 2020 research and innovation programme under grant agreement nยบ 859770.

Labels: MiParea: Respiration, Instruments;methods 

Tissue;cell: HEK 

HRR: Oxygraph-2k, TIP2k, O2k-Protocol 

BEC, SUIT-001 O2 ce-pce D003 

Cookies help us deliver our services. By using our services, you agree to our use of cookies.