Tiling and stitching were performed with Keyence BZ-X Viewer software. Image quantification was performed in ImageJ as shown in fig. S23 . Images were split into component channels. Nuclei (blue channel) were quantified by applying Gaussian Blur (3 Sigma), thresholding, performing watershed calculation for segmentation, and counting. Lipid accumulation was quantified by applying Gaussian Blur (2 Sigma), thresholding, and quantifying the total area above the threshold. The level of adipogenesis, expressed as the adipogenic index, was assessed by dividing the total lipid area by the total number of nuclei to obtain a number with arbitrary units for comparing absolute levels of adipogenesis. Relative adipogenesis was calculated as the ratio of the adipogenic indices in treatment and control samples; here, the control is the untreated sample from the same cell type and BR (for example, ICAM1–TGFβ BR A/ICAM1 control BR A). This ratio represents induction or suppression compared with the baseline.
Gaussian blur
Manufactured by Merck Group
Gaussian Blur is a digital image processing technique that applies a Gaussian function to an image, blurring the details and smoothing the transitions between different areas of the image. This operation is commonly used in image editing, computer vision, and photography to reduce noise, blur edges, and create a soft, defocused effect.
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Lab products found in correlation
2 protocols using gaussian blur
1
Quantitative Image Analysis of Adipogenesis
2
Epidermal Cell Size Quantification
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Images of epidermal cells labeled by Nrg-GFP (1,024 × 1,024 pixels) taken with a 20X objective were first processed by Gaussian Blur (Sigma: 1) and then Auto Local Threshold (Phansalkar method, radius: 30). Isolated particles below the size of 500 pixels were removed by the Particles4 plugin (http://www.mecourse.com/landinig/software/software.html ). The Nrg-GFP signal was then converted to single-pixel-width skeletons of epidermal cell borders using the Skeletonize (2D/3D) plugin. Images were then visually inspected to ensure that all epidermal cell borders were accurately labeled. Any erroneous epidermal cell borders were removed. Regions of interest (ROIs) were manually drawn to encompass the epidermal cells for quantification. Analyze Particles was then used to measure area, perimeter, height (Feret), and width (minFeret) for each epidermal cell in the ROIs. Epidermal cell size ratio was calculated as average cell size in the RNAi-expressing region/average cell size in the WT region.
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