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Brain sections from each treatment group were processed simultaneously for each discrete brain region (n = 1 3 sections per brain region of interest per mouse from n = 5 8 mice per treatment group).
Two successive brain sections from each rat were stained with H&E and TUNEL staining, respectively [ 32].
Images of the relevant brain sections from each individual were taken with a digital camera (Leica DFC320, Solms, Germany) connected to a stereomicroscope (Leica M, Leica IM 50, Solms, Germany).
Hematoxylin and eosin (H&E) staining was performed on at least two brain sections from each mouse to align all brains to approximately 1.3 mm lateral to the midline using a mouse brain atlas [ 52].
26 Three brain sections from each mouse separated by 300 μm, approximately corresponding to sections at bregma coordinates −1.4, −1.7, and −2.0 mm in the mouse brain atlas 25 were used for AT8 staining.
Haematoxylin and eosin stained brain sections from each mouse were scored blind for degree of vacuolation (ranked 0 to 5) found in nine grey matter regions and three white matter regions (Fraser and Dickinson, 1968).
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In addition, for in vivo quantification, cell numbers were estimated from a series of every eighth coronal brain section from each animal.
The thickness of the granule cell layer was measured in three brain sections from three mice of each genotype.
Similar results were obtained using brain sections from three different EGFP-injected animals.
In vivo immunohistochemical analysis demonstrated little caspase-9 activation in the majority of hippocampal brain sections from control brains.
Most of the GFP-positive neurons resided in the expected CP layers in brain sections from embryos of both genotypes.
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