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The fitting procedure for each particle is based on a threshold subtraction and vector gradient analysis for image processing followed by two-dimensional Gaussian fitting (see 'Materials and methods').
After background and threshold subtraction, the values were averaged between several samples of each genotype (considering left and right sides separately, for the number of spinal cord sides used, for each plot indicated in the corresponding figures) to generate an average signal distribution plot.
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An algorithm was designed and implemented in five stages named DSWTS Divisionn, Segmentation, Watershed techniques, Thresholding, Subtraction).
In this study, we used a method which applies a volume-reproducing intensity threshold after subtraction of local background.
The integrated density of the images from cells without laminin treatment was considered 0 and used to establish the threshold for subtraction of background fluorescence in other images.
After threshold background subtraction, the remaining image segments of reasonable sizes are analyzed.
The FDG-PET image of the primary tumor was segmented using the ROVER 3D segmentation tool based on thresholding at the volume-reproducing intensity threshold after subtraction of local background.
where Th is the background subtraction threshold normally used.
C2C12 cells (Fig. 2): For quantification of rapsyn-GFP signals, images were binarised (50 255 greyscale threshold) upon background subtraction.
Software settings were: smooth deactivated; diameter of largest sphere of 0.7 µm; threshold for background subtraction of 1030 µm2; split touching objects enabled; estimated diameter of 0.444 µm; quality threshold above 260; sphericity threshold above 0.550 (94%+ selection).
The mean and 2 standard deviations (SDs) were calculated for all measured values within a slice, and the 2 SD value used to set a subtraction threshold for all pixels in that slice.
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