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Validation of ELISpot

Guest Margot

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Guest Margot


I have developed an ELISpot protocol that seems to be working quite well. Before I apply the protocol to test more of my precious patient samples, I wanted to check if there are (official) rules for the validation of ELISpot assays in terms of intra and inter-assay variability?

I can see some variability between assays, but I suspect this is because my counting may not be that accurate. I now use tryptan blue staining and manual counting to determine viabilty and cells/mL which might not be ideal as it induces variability. What other methods are recommended?

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