Moreover, approximately half of the total nuclei were positive for the Neu-N marker (48.3±3.7% of NeuN-positive nuclei/DAPI total nuclei), indicating that half of the cells are differentiated mature neurons.
DAPI staining shows total nuclei.
Hoechst 33342 nuclear dye was then applied to visualize total nuclei.
A minimum of 3000 nuclei were counted per sample and a ratio between total nuclei with positive nuclei to total nuclei was determined using a minimal and maximal area of 100 and 1000 pixels respectively.
For Ki67 and TUNEL staining, data was represented as a percentage of positive nuclei among total nuclei in each field.
The number of total nuclei per square millimeter was 29.8% and 67.4% higher in 27- and 33-month control rats than the adult animals, respectively (P<0.05).
Cells were nuclear stained with DAPI, and the TUNEL positive and total nuclei were counted under the fluorescent microscope (IX71, Olympus Corporation) in 5 view fields per well.
Slides were stained by immersion in fresh 10% Giemsa stain for 10 min. 2,000 nuclei were counted and percentage of prophase/prophase-like and metaphase nuclei was determined among total nuclei.
Epithelium proliferation, as indicated by the labeling index (the ratio of labeled nuclei to total nuclei in the crypt) is 15.6±1.7 in duodenal ulcer and 17.8±1.5 in duodenitis.
The percentage of dying cells was assessed by counting the pyknotic nuclei over total nuclei into the CA1 layer of lesioned and healthy control animals in serial brain sections (each 200 µm) as described below (n = 3 rats/time point).
After the slides were scanned at low magnification, the cells with positively stained nuclei were counted in six microscopic fields in each tumor and the Ki67 index was calculated as the ratio of Ki67 positive nuclei per total nuclei.
