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Background-corrected maximal pixel intensity (PImaxcorr) in the left ventricle was determined with videodensitometry.
The third row in Fig. 2 finally provides the test images to which independent Gaussian noise of mean zero and a standard deviation of 50% of the maximal pixel intensity was added.
The second row of this figure consists of images where independent Gaussian noise of mean zero and a standard deviation of 10% of the maximal pixel intensity was added to the individual pixels.
Fig. 2 Sets of synthetic (256 times 256 pixel) test images arranged in columns where the first image is the one without noise and the second and third images were created by adding independent Gaussian noise of mean zero and a standard deviation of 10 and 50% of the maximal pixel intensity to the corresponding image in the top row.
Gaussian noise with a mean of zero and standard deviations of 10 and 50% of the maximal pixel intensity was added to the individual pixels of the noise-free images individually to create two more images and thereby complete the sets.
While one of the images in each of these sets was free of noise (and also free of systematic errors so that it was perfectly 2D periodic), independent Gaussian noise of mean zero and a standard deviation of 10 or 50% of the maximal pixel intensity was added to the individual pixels of that image in order to create two noisy images for each set of test images.
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All figure images are maximum projection Z-stacks, with intensity measured as maximum pixel intensity.
The characteristic "flattening" of senescent cells was reflected by the decline in the density of staining (intensity of maximal pixel) of DNA-associated fluorescence [4,6-diamidino-2-phenylindole (DAPI)] paralleled by an increase in nuclear size (area).
The observed decrease in intensity of maximal pixel of DNA/DAPI fluorescence and the increase in nuclear area measured by LSC is a consequence of a change of cell geometry namely its extensive flattening of the senescent cell.
Thus, we studied the effect of exposure of A549 cells to one such inhibitor, TSA, by measuring the changes in intensity of maximal pixel DNA/DAPI fluorescence and nuclear area in the cells growing in its presence (Fig. 7).
Thus, the biomarker based on analysis of nuclear area and intensity of maximal pixel of DNA-associated fluorescence is a sensitive reporter of the characteristic change in cell phenotype considered to be the hallmark of senescence.
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