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In the instance of discrepancy between the two observers, each mouse was counted as a half being in that category.
Immediately after acetic acid injection, the number of writhes per mouse was counted for 30 min.
Fourteen days after the injection of B16-F10 melanoma, the number of lung metastases in each mouse was counted.
Five minutes after the injection of acetic acid, the number of writhing responses per mouse was counted for 10 minutes during acetic acid-induced abdominal writhing [ 7].
The number of apoptotic cells found in 10 complete, midaxially sectioned duodenum crypts of each mouse was counted as an indication of intestinal damage.
The number of writhing movement (painful Muscular contraction) of each mouse was counted for 10 min, starting from 15 min after the injection of acetic acid.
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Non-serial sections were evaluated and 100 axons per mouse were counted.
For assessing proliferation, 940 1400 nuclei per mouse were counted and the "MIB1 index" was determined as the percentage of nuclei positive for MIB1 immunohistochemistry. CD3- and CD68-positive cells were quantified in relation to total number of nuclei.
Twelve sections from each mouse were counted by investigators blinded of the genotype and treatment of the samples for darkly stained nuclei representing c-Fos-like or P-Erk1/2 immunoreactivity within the brain nuclei of interest, which were defined according to the mouse brain atlas [34].
A minimum of 50 BrdU positive cells per mouse were counted in the SGZ and GCL and, as the markers were located in different cell compartments (BrdU in the nucleus and TuJ1 in the cytoplasm), cells were exhaustively analyzed for double staining with tridimensional serial reconstruction.
For quantification of ERα and PR positivity, at least 2,000 epithelial cells per mouse were counted.
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