Seahorse Xfe96 mito stress maximum OCR

I recently ran a mito stress test using the Seahorse Xfe96 and my OCR (pmol/min/cells) values were higher than I typically see for published mitostress assays.

Basal OCR for control and treatments ranged from 150-280 and maximum OCR went up to 580. I am wondering if these values are too high to be accurately measured by the Xfe96 or if these values are acceptable?

Thank you in advance for any help!


  • Hi Rebecca,

    Thank you for contacting Cell Analysis Technical Support.  We recommend maximum basal rates for 96 well platforms to be 160pmol/min. You may have potentially too many cells/well. If you could send your Wave file to, we would be happy to take a look for you! How many cells did you seed per well and what type of cells are these?

    Thank you!

    Courtney Nadeau Watts

    Technical Support Scientist

    Cell Analysis Products

  • Hi Courtney,

    Thanks for your quick response! I do believe we had too many cells per well. We are using rat primary cortical neurons at 80 and 100k cells/ well because in many cases a large portion of the neurons do not differentiate. We had previously done a test of cell density where the basal OCR looked fine at the same cell density but I think between different neuron preps we had different numbers of neurons that successfully differentiated. 

    I am mostly wondering if there is any utility to the data as just a preliminary finding if the basal OCR is high or if the data is totally unusable in that case?

    Thanks again,


  • Hi Rebecca,

    You are very welcome!  It is hard to say without seeing the data, unfortunately. We make the recommendation of 160pmol/min as the maximum because when it starts higher than that, once the FCCP is injected, the cells begin to work so hard that most/all of the O2 in the wells will be consumed, resulting in anoxic cells.  If you would like to send me the data file, I can better advise you!

    Thank you,


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