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After completion of the study, the baby rats were counted and separated by sex with observation for abnormality at the time of weaning.
The morphologically undamaged MNs (i.e. large neurons, with a soma diameter >20 μm and a distinguishable nucleus, similar in appearance to those of the contralateral ventral horn and to those in control rats) were counted in a 10× microscopic field.
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Every sixth 40-μm-thick section of each SN of every rat was counted (from 8to1010 sections per animal, usually nine sections per rat).
The sections were stained with anti-CS antiserum and the number of LS in six sections from each rat was counted.
To determine the time point where the RGCs decreased in aldosterone-treated rats, the numbers of RGCs were counted every week.
As it is not known if these rat loci contain unique sub-loci, human risk associated polymorphisms mapping to overlapping rat regions were counted only once.
G8, G10 and congenic rat cohorts were counted by the same person, but at separate occasions.
Wells were supplemented with OptiMEM with 1% FBS and 10 ng/ml VEGF (R and D systems) and incubated at 37°C, 10% CO 2. Angiogenic sprouts from rat aortas were counted after 4 days and 8 days respectively.
The sequence reads mapping the rat genome were counted, and the differentially expressed (DE) gene values of the two clones were compared using the DESEq package in Bioconductor [ 40].
The embryos were fixed in 4% paraformaldehyde at E16. Rat brain slices and in utero electroporated mouse brains were embedded and cut at 20 µm with a Cryostat All electroporated cells present within the striatum of each section (n = 4 per electroporated rat-slice/mouse brain) were counted.
Rats in groups I-II were counted as "normocholesterolemic rats", while those in groups III-VI were counted as "hypercholesterolemic rats".
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