On the protocol, in the plate map, accidentally I named some wells with cells as background. I have the correct background wells applied, my problem is I added three more backgrounds in wells with cells and I don't know how to obtain the data without the normalization through the background. I would like to obtain my data and do by myself the necessary corrections.
Thanks!
Belinda Lago Vallejo It is not advised to acquire data from a Seahorse mitostress experiment without first performing background normalization to eliminate any non-mitochondrial respiration signal that may add to the data. The existence of background wells containing cells may result in an inaccurate assessment of mitochondrial respiration, which may impact the interpretation of your results.
If you have already performed the experiment with improperly labeled wells, it may be impossible to acquire data without the backdrop normalization. You could, however, attempt manually correcting the data by removing the background signal from the wells containing cells. This would necessitate a thorough grasp of the Seahorse protocol as well as the experimental strategy.
To identify the best course of action, it may be best to confer with someone who has expertise with Seahorse mitostress trials and data analysis. They may be able to offer advice on how to correct the data or recommend alternative methods for obtaining precise findings.
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