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For quantification, 10 high power fields (40×) were randomly chosen under the microscope and the total number of positive cells was counted for each tumor.
Once a region of interest for imaging of the GUVs was chosen under the microscope, ∼500 µl of CLR01 (150 µM) in 10 mM NaH2PO4 pH 7.6, was added.
For quantification of PPARγ expression (Fig. 6D and I), the number of PPARγ-positive cells and Hoechst-positive nuclei in five different fields randomly chosen under the microscope using a 20 × objective was counted, and the portion of PPARγ-positive cells was calculated.
For the quantification of perilipin expression (Fig. 6E and J), perilipin-positive areas in five different fields randomly chosen under the microscope using a 20 × objective were calculated by ImageJ software and the values were divided by the number of Hoechst-positive nuclei.
For quantitative analysis of internalized SPARC of Skm-PCs (Fig. 1E) and itga5 expression (Fig. 2D), five different fields randomly chosen under the microscope using a 40 × objective were photographed and their fluorescence intensities were calculated as positive area by ImageJ software (ver.1.43, NIH, Bethesda, MD, USA), and then, the values were divided by the number of Hoechst-positive nuclei.
Skm-PCs cultured in ADM for 4 days were fixed and stained with Oil Red-O mixture (2 3 mixture of 0.5% (w/v) Oil Red-O (Sigma) in 2-propanol and distilled water) for 10 min. For quantification of adipogenesis, five different fields randomly chosen under the microscope using a 10 × objective were photographed, and the areas stained with Oil Red-O were quantified using ImageJ software.
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For systematic counting, three ocular measuring fields, each with a real area of 0.06175 mm, were randomly chosen under a microscope at a power of × 400 within a section.
For each kidney, 10 microscopic fields (×400 magnification) were randomly chosen under a fluorescence microscope (Nikon, Japan), and the area of renal fibrosis was measured and analyzed using analysis software (ImageJ; National Institutes of Health, MD, USA).
Cells on the lower side of the filter were fixed in 4% paraformaldehyde, stained with crystal violet 0.5% and then counted and photographed by randomly choosing different views under the microscope.
Some chose the microscope as their instrument.
Cells of 5 randomly chosen fields per membrane were counted under the microscope at 200x and statistically analyzed.
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