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We noticed that the mean cell fluorescence levels tend to decrease with time (with a timescale of several hours) 7 8 hours after the beggining of the experiment.
(Here μ is the mean cell fluorescence and σ2 is the variance in F).
However, influence of the measuring approach onto the deduced mean cell fluorescence or onto the reconstituted cell fluorescence distribution in the analyzed population is often not considered.
The following formula was applied where Sind is the mean cell fluorescence after 2-h α-factor induction, Sbasal is mean cell fluorescence before α-factor induction, and Sind_noSZ and Sbasal_noSZ are the corresponding post- and pre-induction signals for strains harboring the appropriate Ste5-SYNZIP along with Msg5 without a SYNZIP fusion.
Nearest neighbor smoothing was performed using GraphPad Prism (20 neighbors on each side to obtain a 4 s moving average, second-order polynomial fit) to generate plots of mean cell fluorescence vs time.
Each sample was first analyzed alone to identify its fluorescence intensity profile and then analyzed with an internal standard to measure their ratio of mean cell fluorescence intensities, which is approximately proportional to their C-value ratio, at least if the GC contents of species are similar.
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The mean cell area and cell fluorescence intensity in five areas and 2(C)) were evaluated as described in previous studies [ 13, 14].
The difference in the mean corrected total cell fluorescence between Tnv-6-treated and control cultures was compared using ImageJ analysis.
The patient-control difference in mean per-cell fluorescence is indicative of the patient-control difference in acetylated α-tubulin protein estimated from Western blot analysis in these cells (Abrahamsen et al., 2013).
b The intensity of staining obtained with anti-P-selectin antibody was quantified as corrected total cell fluorescence (CTCF) (mean ± SEM).
b The intensity of staining obtained with anti-VCAM-1 antibody was quantified as corrected total cell fluorescence (CTCF) (mean ± SEM).
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