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For macrophage density, macrophages were counted on 5 digitally captured non-overlapping ×20 magnification fields in the infiltrated area, and reported as a single value/mm2 in each animal (n = 9).
The presence of CMTMR positive cells was determined on a Zeiss LSM510-META confocal microscope by counting CMTMR positive cells in 10 randomly selected 40× magnification fields in a 40 µm frozen section embedded in cryostat matrix (Tissue-Tek, Sakura, Kobe, Japan) [57].
Tissues were evaluated in 10 random 400× magnification fields in three serial sections and graded negative if less than 10% of the cells in a tumour were immunohistochemically positive.
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For each tumour, microvessels were counted within a 200 x magnification field in the area of highest microvessel density.
The number of mast cells in each mouse was expressed as the maximum number of cells/100× magnification field in quadriceps.
An Axioplan light microscope (Carl Zeiss, Oberkochen, Germany), equipped with an eyepiece graticule (grid), was utilized for quantification of MK numbers, per 20X magnification field, in 5 μm BM or spleen sections.
The number of PECAM-1-positive vascular structures in 40× magnification fields was scored along the border of the ventral disc margin.
We estimated the fraction of β-gal+ cells that were co-expressing Olig2 by analyzing and counting cells in high power magnification fields.
Scoring represents: overall stain intensity and percentage of cancer cells stained in four high magnification fields for each sample.
FOXP3+ T lymphocytes were scored in 5 10% increments as a percentage of total CD3+ staining cells in several high magnification fields.
Invasive cells were counted in five × 10 magnification fields.
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