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Relative fold change values were calculated as: mean fluorescence value for each sample at 37°C/mean fluorescence value for each sample at 4°C.
The cytometer was calibrated for each run by adjusting the wild type control samples to give a mean fluorescence value (arbitrary units) of 145±10 at 675 nm and 445±10 at 525 nm. 10, 000 events (sperm cells) were read for each sample.
For region Chr2.10 mean fluorescence value are not reported because it is very large and a mean fluorescence value has little significance.
Fluorescence was normalized for each measurement by dividing the mean fluorescence value for each frame by the starting (frame 1) mean fluorescence value, and triplicate measurements were averaged with standard deviations calculated after normalization for each frame.
This is due to the fact that the software measures the mean fluorescence value within a designed surface.
Using flow cytometry, a clear shift in the mean fluorescence value was observed when cells were incubated with Blon_2347 and Blon_0375 (Figures 5B D), as well as with Ulex Europeus Agglutinin lectin (UEA).
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A minimum of 100,000 events for each sample were recorded and processed with BD CFlow Software to determine % of fluorescent population and mean fluorescence values.
Horses contributing to the four days were 1, 7, 8 and 16.* mean fluorescence values of 10,000 PBMCs are given in arbitrary units.
HUVECs incubated with serum from RHTN showed increased cell apoptosis (4.43 ± 1.9 versus 7.28 ± 1.5%; p = 0.02) and higher reactive oxygen species (ROS) content (987 ± 181 versus 1231 ± 127 mean fluorescence values; p = 0.01), compared to normotensive subjects.
Fluorescence values of individual cell-mimics were mean fluorescence values of the individual segmented cell-mimics.
Data were analyzed using FlowJo software to compute mean fluorescence values.
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