Metabolic Flux Core

Metabolic Flux Core

The Metabolic Flux Core is a new facility established by Professor Ryan J. Mailloux (Director of the School of Human Nutrition). It includes a Seahorse XFe24 Extracellular Flux Analyzer, BioTek Cytation 5 Cell Imaging Multi-Mode Plate Reader, Oroboros O2K Fluorespirometer, and supporting instrumentation and facilities. This includes centrifuges, microscopes, incubators, and a modernized tissue culture facility. The Flux Core and its supporting facilities are used to interrogate cell metabolism, oxidative stress, and ion homeostasis using purified proteins, isolated organelles, tissues, and cultured primary mammalian cells and cell lines.

Funded by CFI JELF and NSERC RTI

Selected publications:

Grayson C, Faerman B, Koufos O, Mailloux RJ. J Biol Chem. 2024 Mar 11;300(4):107159.

Wang K, Moore A, Grayson C, Mailloux RJ. Free Radic Biol Med. 2023 Aug 1;204:287-300.

Koufos O, Mailloux RJ. Free Radic Biol Med. 2023 Jan;194:123-130

Wang K, Hirschenson J, Moore A, Mailloux RJ. Antioxidants (Basel). 2022 Oct 17;11(10):2043.

Hirschenson J, Mailloux RJ. Free Radic Biol Med. 2021 Aug 20;172:1-8.

Seahorse XFe24 Extracellular Flux Analyzer

The XFe24 is a 24-well metabolic flux analysis instrument that provides quantitative and real time measures of mitochondrial respiration and glycolysis in cultured cells. The system can be utilized to measure changes in mitochondrial bioenergetics and glycolytic flux in almost any adherent cultured cell including cell lines, primary cells, pancreatic islets, and can be adapted for measures using suspension cell lines. The system allows for the measure of all states of mitochondrial respiration and uses fluorometric probes to measure both O2 consumption and glycolysis simultaneously in 24 cell cultures. Protocols have also been adapted to measure the bioenergetics of isolated mitochondria (Funded by CFI JELF).

Cytation 5 Cell Imaging Multi-Mode Plate Reader

The CYT5 system is equipped with fluorometric, absorbance, and luminescent detectors to conduct a wide array of plate-based assays. The instrument is equipped with microscopy capabilities (up to 60x) that can allow for imaging of up to 96 wells and for the visualization of real time changes in fluorescence, absorbance, or luminescence in cultured cells or isolated organelles. The system is also plugged into the XFe24 to allow for the correction of respiration and glycolytic flux in the Seahorse with the number of cells per well in the XFe24 plate. (Funded by CFI JELF)

Oroboros O2K Fluorespirometer

The O2K system utilizes an electrode-based system for the high-resolution measure of mitochondrial respiration using either suspension cells, muscle fibers, tissue homogenates, or isolated mitochondria. The system is also equipped with a fluorometer that allows for the simultaneous measure of other mitochondrial parameters, such as ROS production, calcium levels, or membrane potential, using commercially available reporters. (Funded by NSERC RTI)

Metabolic Flux Core List of Equipment, Fees, and Subscriptions

NOTE: Professor Ryan J. Mailloux and his lab are the principal users of this equipment.

Rules for use:

• All users must receive training from the Mailloux lab prior to use of this equipment.
• Access to this equipment is only provided by Professor Mailloux.
• For subscription use, days of the week that it can be accessed must be negotiated with and approved by Professor Mailloux and his lab group.
• Note that scheduling to run an experiment or use for an hour or day must be negotiated with and approved by Professor Mailloux.
• Users must sign up to use this equipment in advance (a week to two weeks prior to the experiment).
• Note that the Mailloux lab WILL NOT SUPPLY you will any reagents or consummables for running your assays.
• Failure to follow the rules or pay the fees in a timely manner will result in automatic loss of of authorization to use this equipment.

TERMS OF ACKNOWLEDGEMENT - USE OF FLUX CORE