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Fluorescence intensity in individual cells was normalized with the mean background intensity obtained in three randomly chosen blank areas42.
The mean background intensity from control and anti-E-selectin-targeted animals was subtracted.
Labeling was considered positive when the intensity of the fluorescent signal exceeded the 95% confidence interval of the mean background intensity.
From these scans, we obtained a relative transmissionaTrel(red dashed line in Figure 1) by normalizing the measured intensity Im with the mean background intensity Ib from regions without particles: Trel=Im/Ib.
Global background subtraction was performed by subtracting the mean background intensity plus two standard deviations from the signal of all experimental spots.
A vessel was considered positive if mean VCAM-1 intensity was higher than mean background intensity, whereas it was considered negative if mean VCAM-1 intensity was below.
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Each result was corrected by subtraction of mean background intensities for each channel.
Metrics for all sample hybridizations including scaling factors, mean background intensities, and percent present calls have been provided (see Additional File 1).
GenePix 6.1 calculates SNR as the ratio of the mean net signal intensity (mean background pixel intensity subtracted from the mean foreground pixel intensity) over the standard deviation of the background pixel intensity.
Firstly, the spot intensities were corrected for local background (mean spot intensity minus mean spot background intensity).
Signals were only assessed as positive if mean spot pixel intensity was higher than the mean local background intensity plus twice the standard deviation of the mean local background pixel intensity.
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mean background pixel
mean background count
mean humor intensity
mean background illumination
mean fluorescence intensity
mean background flow
mean background value
mean fluorescent intensity
mean staining intensity
mean light intensity
mean background subtraction
mean background fluorescence
mean background noise
mean background proliferation
mean background level
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