Dear Yann Dos Santos

instead of setting values to 0 (which will influence your statistics and likely is going to introduce an artefact, you may consider rerunning your samples with our 48-plex cytokine assay with absolute quantification. As it is a qPCR based readout, you have much better sensitivity.

https://www.olink.com/products-services/target/48-cytokine-panel/

You can check the independent comparison vs. Luminex here, in brief the PEA assay not only guarantees better sensitivity (and specificity due to dual antibody binding), but also a much broader dynamic range for different cytokine analytes, which should help especially when you have large variation within your experimental groups.

https://www.olink.com/application/multiplex-analysis-of-inflammatory-proteins-2/

If you have 1 microliter of plasma left, it might be worth a try.

All the best & good luck with your experiment,

Michael

Hello,

Thank you for this interesting proposal, however, we have no more funds for this project... I (finally) found an interesting article that proposes to use fluorescence instead of concentration. This way, I don't have any missing values and it even seems to be more powerful statistically.

If anyone has any clarification on this?

thank you and have a good day,

Yann, Ph.D. student

Hello,

I have come across multiple articles where the

Incase of

Hello Noha Mousaad Elemam , Kalule Wilberforce

Thank you for your answer. The main problem with this approach is that I have very little value for certain cytokines and I can't do good statistics ... And as I previously said, I have no more funds to reconduct the experiment and I have to do with this data...

I think that the use of fluorescence intensity is a good way in my situation, it's relative quantification but statistically powerful.

But thank you once again and have a good day.

Yann

We recently ran into the same issue with ours as well.

We ended up using the standards to create a regression, then reverse predicting the actual values from the samples MFIs. This allowed us to report actual concentrations for the stats we needed.