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Automated segmentation (Spot Detector, Olivo-Marin, 2002) and tracking (u-track, Jaqaman et al., 2008) were used to quantify motility to support this conclusion.
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Delta2D offers a hybrid between segmentation and modeling: starting with segmentation, spots are modeled as Gaussians, and their nonoverlapping boundaries are derived from the models (Fig. 9).
Each data point is the average value of mRNA/cell determined by automated image segmentation and spot counting (N = 60 cells, error bars are SEM).
The scale-25, shape-0.1, and compactness-0.5 used in segmentation of SPOT-5 images fulfilled the condition of segregating two adjacent features.
Ratios are then computed based on explicit segmentation of each spot.
Spot segmentation and background subtraction were based on hybrid segmentation algorithms previously published by our laboratory [ 24, 25].
The software allows the user to verify the accuracy of the spot segmentation against the original fluorescence intensity data.
The GenePix 5.0 image analysis software (Axon Instruments, Inc., Union City, CA) was used for spot segmentation and intensity calculations.
The segmentation of single spots was done semi-automatically via the command 'Find Maxima' and by hand with the pencil tool and a line thickness of 1 pixel.
Spot detection, segmentation and matching followed a strict protocol to ensure a high level of correctness.
The segmentation of cDNA microarray spots is essential in analyzing the intensities of microarray images for biological and medical investigation.
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