I've been using the Seahorse XFe96 with the LCFA Oxidation Test kits and have an anomaly in one replicate with one of my groups having a massive OCR (200-500pmol/min). I remember having some confusion with this replicate regarding which orientation the plate goes into the machine so have looked at a question regarding this on this forum. This replicate gives the plate orientation as 'K', where as my other 2 replicates give the orientation as 'V' - what does this mean in this context? Is the output data still associated with the correct well and group i.e. the OCR and ECAR for well B2 was measured from well B2 during the experiment? Or would this data originate from well G11? I presume since the data knows the orientation of the plate the data is associated with the correct well it was measured in. In this case, I would need to change around my cell counts to normalise each well correctly?
Sorry for the long question, thanks very much!