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Caroline

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  1. Dual ELISpot

    Dear Christian, Thank you very much for your answer. Unfortunately we don't have the new mabtech reader available on site. I started my first comparison experiment last week and it's very difficult for a beginner to decide which gain etc to put on the machine. I tried to compare my ELISPOT and fluorospot counts (IFNg Elispot and IFNg/IL-17 Fluorospot) and there is a much high number of spots with the Fluorospot. I tried to correct them manually but there still very high and it is honestly quite difficult when it is super bright to make a difference between 2,3,5 spots. I have a few questions : - Do you usually recommand not to correct the spots counts and to leave the machine countings (apart from the obvious artefacts ?) - Do you know if there is a way to find tutorial for manual correction (I didn't find any) ? - Can the coating of the plate have an impact on this ? - What about auto-fluorescence ? I read my two cytokines plate on a machine set up for 3 CK and still had some spots on the third cytokine whereas there was no staining there .. - Regarding the Il-17 control what would you consider as a good positive number of spots for this CK ? I have attached the general results of my ELISPOT but happy to discuss more detailed wells if you think it's relevant. - The controls are 4 A-D and H, same for 8 and for 12. - The first four colums are cells with 48 hours incubation before transfering to the plate. - The 4-8 colums are after 24hours in incubation before transfering to the plate - The 9-12 colums are samed day transfer to the plate Thanks a lot Best wishes Caroline IL 17 Cleaning deep636851713438585698.docx
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