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Protocol publication

[…] For image acquisition, we used a high-performance laser-based confocal imaging system (INCell 6000, GE Healthcare, Chicago, IL). For non-spotting experiments, a total of 27 random images were taken for each condition from three independent experiments. For spotting experiments, images were taken from spotted areas in three independent experiments. Cell counting was performed using CellProfiler (). To count the cells, we first counted the number of DAPI-positive cells, followed by counting the number of cells expressing the marker of interest. Student's t-test (comparing two groups) was used for statistical analysis. Co-localization analysis was performed using the WCIF-ImageJ software package and the Image Correlation Analysis plugin (). The plugin uses Pearson’s Correlation Coefficient (PCC) for quantifying the correlation between two channels, as well as calculating Mander’s Overlap Coefficient (MOC). […]

Pipeline specifications

Software tools CellProfiler, ImageJ
Application Microscopic phenotype analysis
Organisms Homo sapiens