In December 2014, the University of Manchester collaborated with DWS to create a scientific poster (Seahorse i2 and H35 combination poster.pdf) about the work carried out in their Whitley H35 Hypoxystation and Whitley i2 Instrument Workstation. This combination of workstations enabled the researchers at Manchester to prepare their cells lines in the H35’s hypoxic environment and then transfer the cells through the transfer tunnel into the i2 (housing their Seahorse Bioscience XFe96 Extracellular Flux Analyzer) without exposure to oxygen. The poster was presented in Seville, Spain.
The i2 was developed in response to an increasing number of enquiries from scientists who wanted to use Seahorse XF Analyzers in hypoxic conditions and were dissatisfied with the solutions available. The i2 maintains an internal temperature no higher than 28°C, excludes carbon dioxide, and provides precise oxygen control – perfect conditions for operation of the Seahorse XF. However, the cells lines need to be prepared in hypoxic conditions so DWS designed flexibility and customisability into this system so that the i2 can be combined with any other Whitley Hypoxystation: from the H35 to the huge H135 HEPA.
The i2 features an incubator above the airlock that allows media to be pre-conditioned in a reduced O2 atmosphere. It also has an airlock on the left-hand side of the system to allow items to be removed without having to be passed back into the H35.
Jonathan Gilthorpe, from Umeå University, Sweden, is currently using the combination of the H35 and i2 for his research and kindly gave us a comment about his experiences:
“Everything is working very well and after a year we are using the H35 for routine growth of primary fibroblast and iPSCs cells under low O2 tension (2-5%). We can passage sensitive cell lines that are prone to senescence due to oxidative stress. Having the i2 connected to the H35 gives us the flexibility of running Seahorse experiments under controlled conditions where the cells are never exposed to a hyperoxic shock. However, we can also run the Seahorse XFe96 under other O2 concentrations as a stand-alone unit in the i2. It gives us the best of both worlds, so am very pleased that we went for this configuration.”
Ultimately, the combination of the H35 and i2 is ideal as two atmospheres are needed for Seahorse hypoxia studies because cells need to be incubated in both O2 and CO2, whereas the Seahorse needs to be in an ambient temperature and O2 controlled environment.