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Each specimen was assigned a score for staining intensity (values 1 4, with 4 representing the most intense stain) and a score for percentage of positively stained cells (values 0 6, with 6 representing greater than 75%).
The staining intensity was scored on a semiquantitative four-point scale as follows: 0, equivalent to the negative control; 1, weak cytoplasmic stain slightly darker than the negative control; 2, moderate stain defined as the intensity between score 1 and 3; 3, intense stain equivalent to or darker than the positive control.
The intense stain masked our ability to image pSmad2/3 in the nucleus.
However an intense stain was evident, in median body, on the periphery of the ventral disc and flagella (Figure 1B).
Caudally, small focal zones of intense stain appeared, with very little immunostaining around them (Figure 2B, top).
The immunostain was evaluated semiquantitatively using a modified Allred scoring scale [14] that takes into consideration the intensity of the immunostain on a scale of 0 3+, with 0 representing no detectable stain, 1+ minimal stain, 2+ moderate stain, and 3+ representing intense stain.
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Scores for the intensity of cell staining were as follows: 1 (mild intensity), 2 (moderate intensity) and 3 (intense staining).
Additionally, the intensity of positive staining from weak to very intense staining was recorded.
The intensity of positive staining from no staining (0), weak to very intense staining was recorded.
Immunohistochemistry for nitrotyrosine and poly (ADP-ribose) synthetase showed an intense staining in the inflamed colon.
Sometimes, more intense staining is observed when the probe is attached to a fluorescent particle.
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