Leo 2024 MitoFit

From Bioblast

MitoFit Preprints         MitoFit Preprints        
Gnaiger 2019 MitoFit Preprints
Gnaiger MitoFit Preprints 2020.4
        MitoFit DOI Data Center         MitoPedia: Preprints         Bioenergetics Communications

Leo 2024 MitoFit

Publications in the MiPMap
Leo E, Rychtarova L, Garcia-Souza LF, Γ…sander Frostner E, ElmΓ©r 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

Β» MitoFit Preprints 2024.4.v2.

MitoFit pdf

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


Version 2 (v2) 2024-06-20
Version 1 (v1) 2024-06-13 - Β»Link to all versionsΒ«

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.

β€’ Keywords: high-resolution respirometry, small-volume module, fibroblast, platelets, isolated mitochondria, 0.5 mL O2k-chamber β€’ Bioblast editor: Tindle-Solomon L β€’ 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: ORCID.pngLeo Elettra, ORCID.pngRychtarova Lucie, ORCID.pngGarcia-Souza Luiz F, ORCID.pngAsander Frostner Eleonor, https://orcid.org/0000-0001-7901-1826Elmer Eskil, ORCID.png Gnaiger Erich

Data availability

Original files are available Open Access at Zenodo repository: https://10.5281/zenodo.11634408
Cookies help us deliver our services. By using our services, you agree to our use of cookies.