Hi everyone, I'm encountering a problem in fixing the cells after the Glyco and the Mito stress assays in Seahorse XFe96 analyser. I start with monolayer of 100% confluent before the assay and after the assay most of the cells are detaching from the wells. And that eventually this affects the measurements of OCR and ECAR. I'm working on adherent cell line. Did any of you noticed this before?
Hi Georgina, I am using a XF24 with adherent cell lines, the same thing happens to me. I would be interested to know if there is a fix, or if we should be using the cell tak to make sure they stay down!
Hannah
I had the same issue with HEK293T cells but after coating the plate (poly-D-Lysine or Cell-Tak) prior to seeding and reducing the mixing to minimum during the seahorse assay I retain nearly all cells - and then I normalize to cell number post assay.
Hi Georgina Barnabas
I used to face a similar problem while working with human monocytes, which are semi-adherent in nature. Eventually, I figured out that it was due to pipetting. You need to change the medium via slow pipetting and not@ touch the pipette tip to the well bottom. With practice, you will stop losing cells.
Thanks.
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