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Background pixels were subtracted from each image, and smoothed droplet perimeters were determined through an arbitrary but consistent contrast threshold procedure.
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Mean local background pixel intensities were subtracted from mean spot-pixel intensities for both channels, which were then combined as Log2-ratios.
The ECL signal was quantitated using a scanner and a densitometry program (Scion Image; Scion, Frederick, MD). Background pixel values were subtracted, the signal was normalized to the amount of actin and the normalized band intensity values were plotted.
GFP intensity was measured as the pixel density in the entire cross-sectional area of each worm from which the background pixel density was subtracted (90 worms per condition).
Background pixel volume was subtracted from each protein band.
The median pixel intensities of individual features were determined using GenePix Pro version 6.0, and background values were subtracted.
Background CARS and TPEF pixel intensity were subtracted from the average pixel intensity of CARS and TPEF signal to obtain CARS signal for fat stores and TPEF signal for autofluorescent granules, respectively.
For both, background signals were subtracted.
Background counts were subtracted from these values.
The background luminance values were subtracted.
Channel backgrounds (median) were subtracted.
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