Leo 2024 MitoFit
| Leo E, Rychtarova L, Garcia-Souza LF, Åsander Frostner E, Elmér E, Gnaiger E (2024) High-resolution respirometry in a small-volume chamber. MitoFit Preprints 2024.4.v2. https://doi.org/10.26124/mitofit:2024-0004.v2 |
» MitoFit Preprints 2024.4.v2.
High-resolution respirometry in a small-volume chamber.
Leo Elettra, Rychtarova Lucie, Garcia-Souza Luiz F, Aasander-Frostner Eleonor, Elmer Eskil, Gnaiger Erich (2024) MitoFit Prep
Abstract:
- Version 2 (v2) 2024-06-20
- Version 1 (v1) 2024-06-13 - »Link to all versions«
Investigating respiratory fluxes is decisive for understanding the complex interplay between metabolic processes. Studies of cells and tissues with limited sample availability and low respiratory rates may benefit from small experimental volumes. We examined if the 0.5 mL chamber yields results consistent with the 2.0 mL chamber at identical sample concentrations used in high-resolution respirometry with the Oroboros O2k.
The background O2 flux was 4-fold higher in 0.5 mL than 2.0 mL chambers at air saturation, but was reproducible allowing for accurate background correction. The optimal stirring speed was between 550 and 750 rpm in the 0.5 mL chamber.
Respiratory fluxes in living cells, permeabilized cells, and isolated mitochondria were measured in parallel in the 0.5 mL and 2.0 mL O2k-chambers, using SUIT protocols with 14-20 titrations. O2 fluxes in the two chamber types were identical within limits of detection. The 0.5 mL O2k-chamber, requiring close to four times less sample than the 2.0 mL chamber, offers a significant advantage for studies with limited amounts of sample or low respiratory capacities.
• Keywords: high-resolution respirometry, small-volume module, fibroblast, platelets, isolated mitochondria, 0.5 mL O2k-chamber • Bioblast editor: Tindle-Solomon L • O2k-Network Lab: AT Innsbruck Oroboros, CZ Prague Zeman J, SE Lund Elmer E
Labels: MiParea: Respiration, Instruments;methods
Organism: Human, Mouse
Tissue;cell: Heart, Nervous system, Blood cells, HUVEC, Platelet
Preparation: Permeabilized cells, Homogenate, Isolated mitochondria, Intact cells
Regulation: Coupling efficiency;uncoupling, Cyt c, Uncoupler, Fatty acid Coupling state: ET, LEAK, ROUTINE, OXPHOS Pathway: F, N, S, Gp, CIV, NS, Other combinations, ROX HRR: Oxygraph-2k, O2k-Protocol
ORCID: 
Leo Elettra, Rychtarova Lucie, Garcia-Souza Luiz F, Asander Frostner Eleonor, 
Elmer Eskil, Gnaiger Erich
Data availability
- Original files are available Open Access at Zenodo repository: https://10.5281/zenodo.11634408
