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However, since focality of expression was a relatively subjective observation dependent on variable factors such as the even distribution of antigen at the time of staining, we grouped the results into better definable groups based only on the strength of staining.
Subsequently, strength of staining is associated to each cell through nuclear segmentation and region-based tessellation.
For 7 of these markers, strength of staining of each core on the TMA was formally scored.
Intensity assessed the strength of staining and was scored as 1+, weak; 2+, moderate; and 3+, strong staining.
Subsequently, the strength of staining is quantified by a color decomposition model that is optimized by a graph cut algorithm.
The percentage of positive nuclei within tumour tissue was multiplied by the strength of staining: 0 (negative), 1 (weak), 2 (moderate) and 3 (strong).
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However, differences in reactivity – chromatic strength and localisation of staining – were noted with the IP-10 and IP-14 antibodies.
Recently, our group has also demonstrated that Trop-2 expression is significantly higher in endometrioid endometrial carcinomas (EEC) compared to normal tissues representing both proliferative and secretory phase endometrium, and that strength of immunohistochemistry staining correlates with increasing grade.
On a semiquantitative scale of staining strength the sections varied from weak (11), moderate (5), strong (8) to very strong (8).
Scores were graded in a blinded fashion, and the criteria assessed were (1) proximity of CD31 staining to the joint, (2) strength of CD31 staining, (3) degree of infiltration of synovial cells into the joint and (4) overall status of joint architecture.
The strength of the staining intensity was examined using the following range: 0, no cell staining; 1+, mild staining; 2+, moderate staining; and 3+, strong staining.
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