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Each of the macaques generated strong responses against all three antigens, with a minimum ELISpot count of over 500 and a maximum above 2100 (mean spot count 1050±662 SEM).
The mean spot count for the experimental wells was varied over 40 values from a mean of 10 to 50 per 100,000 PBMCs.
The mean spot count for the background wells was set at 10 per 100,000 PBMCs, reflective of the mean in our example data set.
For each medium, the mean spot count from labs that did an overnight rest was larger by more than 35 spots compared to the mean spot count from labs that did not do an overnight rest.
One commonly used alternative hypothesis is that the mean spot count in the experimental wells is greater than that of the background or control wells (a one-sided alternative hypothesis).
Similar(55)
To assess the overall true positive rate, background spot counts for each donor were again generated from a Poisson distribution with background mean spot counts of 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50; however, the experimental means were shifted by 6 (small difference), 20 (moderate difference), or 50 (large difference) relative to the background means.
A common null hypothesis in the ELISPOT response determination setting is that there is no difference between the average (mean) spot counts in the experimental and control wells.
I mean, spot the difference….
Responses were measured as specific spot forming units (SFU), calculated as the difference between the mean treatment spot count and the mean negative control spot count.
A detectable response was defined as a mean treatment spot count that was more than 10SFU 105 cells−1 greater than twice the mean negative control.
This is illustrated in Fig. 3 where the mean medium spot count for all reported replicates is plotted on the vertical axis with its corresponding rank displayed on the horizontal axis.
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CEO of Professional Science Editing for Scientists @ prosciediting.com