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For the manual readout the plaques were counted by eye in separate experiments.
The results of the automatic readout and what would be the corresponding manual readout within the same experiment are thus closely similar, if not identical.
We compare this assay with manual readout and with other neutralization assays using a reference panel of inhibitory reagents, i.e. plasma and monoclonal antibodies.
Further examination of this latter group revealed that in 11 out of 14 combinations, the automatic readout was more sensitive than the manual readout in detecting neutralization (denoted as triangles above the diagonal line in Figure 5).
Similar(56)
Taken together, the APR assay detected neutralization similarly, but with generally increased sensitivity, as compared to the manual PR readout.
These 14 points were not the result of an image analysis artifact, since no false positive background was detected with the negative plasma and the manual and automatic readout of other inhibitory reagent-virus combinations run on the same plate and day were in close agreement.
We compared the manual and automated readouts using 72 virus-plasma combinations, including eight combinations with HIV-negative plasma.
In the remaining 14 virus-plasma combinations (19%), variation of IC50 was more than 3-fold between manual and automatic readouts.
In 46 cases (64%), the manual and automatic readouts of 50% inhibitory concentration (IC50) were within twofold of each other and in 12 cases (17%) the difference was within a 2- to 3-fold range.
Traditionally, the identification of gene products and neurons involved in behaviors such as thermosensation has relied on manual assays with a single readout - for example, a thermotaxis index measuring how well animals navigate toward the cultivation temperature.
At the very least, a good manual assessment will alert the assessor to any spurious readout from a computer model.
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