Cytokine staining in IMC

[Paulan]

Hi,

Has anybody tried to detect cytokines (besides IL1b) using IMC? I'm assuming it's difficult but wondering what people have tried and if anything has worked for them so far. Any indication that it does not work is also very welcome.

Thanks,
Paula

[dianebender]

Hi Paula,
Our group has imaged TNFa, IL-6, IFNg and IL-10 in human bone marrow or spleen using IMC. The IL-10 was a Fluidigm CyTOF antibody cat#3166008B used at 1:50 following the Fluidigm suggested AR protocol for FFPE. The other antibodies were not Fluidigm.

Diane

[Paulan]

Hi Diane,

Thanks for your reply! Good to hear that it can be done. Would you say the cytokines detected were mostly cell associated/intracellular or did you also detect released cytokines in the tissue?

Cheers,
Paula

[dianebender]

Hi Paula,
The cytokines were cell associated/intracellular.

Diane

[XMUCyTOF]

Hi,
Do you need an inhibitor to stop the cytokine secreting from the intracellular?

[Paulan]

Hi Diane,

Thanks for your reply, sorry I forgot to check back but good to know they are intracellular/cell associated!

Cheers,
Paula