Seahorse XFe24 Analyser: Live‐cell analysis
||Overview: The Seahorse XFe24 Analyser measures the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of live cells in a 24‐well plate format. By combining these measurements with chemical intervention assays, selected cellular functions can be investigated in real time including mitochondrial respiration, glycolytic function, cellular ATP production and immunological activation. Both cultured cells and ex‐vivo samples can be used and our system can also be run under normoxic and hypoxic conditions (see below). Further details, including the types of experiments that can be performed and video tutorials can be found on the Agilent website. A useful resource is also Agilent’s cell reference database where previously published experiments can be searched by cell type.
Management of the system: The Seahorse XFe24 Analyser is managed by the Mass Spectrometry Research Facility, in the Department of Chemistry. It is available to all university research groups, with priority given to PI’s involved in the original instrument application and other Wellcome Trust funded researchers. Access is organised in two ways: The first is collaboratively (with the McCullagh group for standalone Seahorse analysis or combined with mass spectrometry‐based metabolomics). The second is on a service basis. In both cases the preferred format is for the Seahorse system to be run by a member of the McCullagh Group alongside the collaborator where possible. For expert users, direct access can be arranged after discussion in the first instance. In order to support instrument upkeep and maintenance there are charges for use (see below for details).
Accessing the Seahorse: All Wellcome Trust Seahorse grant co‐applicants and other Wellcome Trust funded researchers will be given priority access. For those wishing to use the system collaboratively please contact James McCullagh or if service‐only access is preferred, please contact John Walsby‐Tickle. In both cases an initial discussion will be arranged to find out the aims of the project and establish which Seahorse assays are required. Please note: Wave software used for planning and analysing Seahorse experiments can be downloaded free.
Cost of using the Seahorse: The default approach is for the McCullagh Group to provide all plates and consumables (charged at cost), although its fine to use your own if you prefer. The cost of consumables and maintaining and running the system, will be recovered via a charge‐for‐use model. The cost for co‐applicants and collaborators is £90 per plate (or part thereof) analysed (£50 if own plates and consumables used). The cost for service access is £180 per plate (or part thereof) (£130 if own plates and consumables used). A purchase order request for payment will be sent to the PI of the lab when the experiments are completed.
We are happy to be included as collaborators on grant applications/write letters of support etc.
Hypoxic and anoxic experiments: The Seahorse has been setup within a glove‐box system to enable optional experiments in a controllable anoxic or hypoxic environment when required. If you would like to use these capabilities please discuss directly with James McCullagh in the first instance.
We look forward to working with you on live cell metabolic analysis.