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A is the total area of a well (28.27 mm2), Ami is the area of the microscopic image (0.015552 mm2), Cn is the average cell number per microscopic image and Df is the dilution factor per millilitre of reactor content.
(C) Average cell number of H2B-GFP positive cell in the control (n = 41), AurkB-OE (n = 30), AurkC-OE (n = 32), siAurkB (n = 40), and siAurkC (n = 31) groups, each point represents data from one embryo, the bar and whiskers indicate means and SD, H2B-GFP positive cells are the daughter cells of the injected blastomere.
Clone sizes were quantified based on the number of cells/nuclei per clone (observed by DAPI staining) and average cell number per clone and standard deviations were graphed using Microsoft Excel for both clones and twin spots.
To generate this average cell number per droplet, we estimated the density of the mixed culture under microscope using a Petroff-Hausser counting chamber and then diluted accordingly before injecting it into the device.
The average cell number yielded per passage during in vitro culture ranged from 3.5×106 at p1 p5 to 7.5×106 at p6-p10 to 107 at later passages (p11 16), suggesting that culturing resulted in an increased proliferation rate (Figure S5).
Total cell number (N) was calculated using the formula N = Nv x Vref, where Nv is the average cell number per disector volume (corresponding to 15×15×40 µm3) and Vref (reference volume) is the total volume of the dentate gyrus.
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Average cell numbers and standard deviation were calculated using Statlab (SPSS Inc, Chicago, Illinois, USA).
Cells were counted after 5 days and the average cell numbers from triplicate measurements were plotted.
Cells were counted each day and the average cell numbers of triplicate wells were plotted.
To account for differential proliferation, viable cells were enumerated at each time point and changes in metabolite concentrations normalized based on average cell numbers.
Fig. 3 Average CaCo2 cell number following 72 h of free and SMA Ral treatments.
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