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ELISpot cell dulitions

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Hi all, I understand you often have to plate multiple cell dilutions in the ELISpot to determine the appropriate number of cells that will produce good quality spots. My question is, if all your cell dilutions produce spots, how do you go about deciding which spot counts to use? Do you average them all? Which cell dilution do you "believe"? Thanks!

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It is smart to begin your project doing some titrations to figure out an optimal cell number for your particular antigen. Maybe responses are much better at 400K cell/well compared to 250K/cells. This is difficult to know prior to actually testing. 

However, once you have found a cell number where you dont feel limited by a lack of "cell-to-cell" contact, you can stick with that cell number for an entire study. 

ELISpot and FluoroSpot readers typically have good linearity on counting spots from 0-600 spots. With Mabtech IRIS we see good linearity between 0-2000 spots even beyond. As a result, you are not limited by the read out equipement for capturing both weak and very strong responders. 


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