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ASCs were shown to persist within a defect site for two weeks (shown by sex chromosome analysis and quantified using Luciferase+ ASCs).
RNA expression was detected by Northern blot analysis and quantified using a Molecular Dynamics Storm 860 PhosphorImager/Fluorima. Protein expression detected by Western blot analysis was quantified using the ImageJ program.
Radioactive signals were identified by Typhoon 9410 (Amersham) analysis and quantified using ImageQuant 5.2 software.
Gels were exposed to phosphorimager analysis and quantified using the Bio-Rad Image Quant software.
Samples were harvested at 10 hours following the respective treatment and lysates were prepared for western blot analysis and quantified using the actin loading control for normalization.
The femurs were removed at death, prepared for histomorphometric analysis, and quantified using a computer-aided image analysis system (Bioquant II, R and M Biometrics, Nashville, TN) as previously described (Simic et al, 2006).
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Gene expression was analyzed with Stratagene analysis software and quantified using the 2−ΔΔCt method.
After 6 h of incubation at 37°C, the formation of capillary-like networks was examined under an inverted microscope (Zeiss, Oberkochen, Germany), equipped with charge-coupled device optics and a digital analysis system, and quantified using the Angiogenesis Analyzer toolset (26) for National Institutes of Health ImageJ software.
Immunoisolated proteins were detected by Western analysis using anti-GFP antibodies and quantified using the QuantityOne software (Bio-Rad).
They were then analyzed by the Bioanalyser Analysis System (Agilent, Santa Clara, USA) and quantified using real time PCR.
Apoptotic cells were detected by FACS analysis (Becton-Dickinson, NJ, USA) and quantified using cellquest software (Becton-Dickinson).
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