Slides were scored on a scale of staining intensity, with 0 representing no staining, 1 representing weak staining, 2 representing moderate staining, and 3 representing strong staining.
On a semiquantitative scale of staining strength the sections varied from weak (11), moderate (5), strong (8) to very strong (8).
A final hybrid score (H) was assigned to each sample, based on the product of a 0 3 scale of staining intensity and of the percentage of positive cells (0 100%), with a possible range of results from 0 to 300.
For scoring Lgr5 and MMP2 expression, immunohistochemical staining of cells was evaluated according to a score that added a scale of intensity of staining to the proportion of stained cells (Xi et al, 2010).
Under light microscopy (Olympus), the distribution of phospho-LYRIC in the nucleus and cytoplasm of the cells was rated according to a score that determined the scale of intensity of staining to the area of staining using ImageMaster software.
To establish a scale for intensity of staining an initial pilot group of sections was examined to observe the range of staining.
The intensity of staining (scale 0 4) and the percentage of stained cells were evaluated.
Immunoreactivity for SNCG in tumour cells was graded as either negative or positive according to a four-value classification scale as follows: area of staining as <10percentt (0) or >10percentt (1) of all cancer cells stained within the section, staining intensity (>10% of all cancer cells stained within the section) was graded as weak (1), moderate (2), or strong (3).
Cells were scored for intensity of staining on a scale of 0 (no staining), 1 (weak staining, light yellow), 2 (moderate staining, yellowish brown), and 3 (strong staining, brown).
