Phenotypic Screening is used to characterise the effect of test molecules by quantifying morphological/phenotypic changes in cell-based models.
A scratch was formed on a A549 cell monolayer and treated with Olaparib (Ola), 5-Fluorouracil (5-FU); a combination of 5-fluorouracil and Olaparib (5-FU + Ola). Images were acquired using live imaging platform at the indicated time points (hrs). The wound size/scratch was measured at all time points. The wound closure percentage was analysed assuming 0% closure at 0 hours, and calculating all the following time points as ratio to 0 hours [ *p<0.05; **p<0.01;***p<0.001; ± SEM].
Monocytes (CD14+) were isolated from PBMCs of healthy donors followed by treatment with M-CSF for 6 days. On day 6, the naïve macrophages were exposed to GM-CSF + IFNγ and M-CSF to generate M1 and M2 macrophages (MΦ) respectively. The cells were then exposed to fluorescent zymosan particles (green) and incubated for 3 hours. After 3 hours, the cells were washed, stained with fluorescent membrane marker (red) and DAPI (blue) for nucleus.
The images above showed M2 macrophages engulfed higher numbers of zymosan particles when compared to M1 macrophages (Objective= 20X).
Percentage change in gel contraction upon pre-treatment with 2,3 BDM followed by incubation for 4 hours with Carbachol. For each treatment the percentage change in the gel contraction was calculated based on the values obtained at time 0. Each condition is representative of 3 biological replicates (n=3); mean+SEM. Data analysis was performed using the 2-WAY ANOVA analysis, *p<0.05; **p<0.01, ***p<0.0001; Carbachol vs. test ingredients groups.
The target Raji cells were pre-treated with or without Rituximab at different concentrations and stained with pHrodo Red Cell Labelling (red). The THP-1 derived macrophages were stained with calcein staining (green). To assess antibody-dependent cellular phagocytosis of target Raji cells by THP-1 derived macrophages the two cell types were co-cultured for 6 hours. Data is represented as the red fluorescence intensity (RFI) at 0 hours vs. 6 hours in all treatment groups.
Normal Human Bronchial Epithelial Cells were grown on Air Liquid Interface (ALI) and allowed to differentiate for 28 days. At the end of the culture the cells were fixed and embedded into paraffin. The paraffin blocks were sectioned at 5µm thickness using Microtome and fixed onto slides. Haematoxylin and Eosin (H & E) staining were performed and slides were imaged at 4, 20 and 40X magnification. Distinct goblet and ciliated cells were observed on Day 28 of the ALI differentiation
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