CD34 was assumed to stain specifically the endothelial cell cytoplasm and therefore the value acquired from the morphometric count was considered to represent the percentage of the total microvessel area of the tumor sample.
Compared to the direct counting of AgNOR signals, morphometric analysis allows a more objective and reproducible quantification on histological sections [ 17, 32].
We developed an objective morphometric analysis of nuclei size and irregularity [ 41], which showed that RT treatment increased the percentage of irregular nuclei, characteristic of MC, when compared to the individual treatments, in agreement with direct counting of irregular nuclei.
Counting of neurons, oligodendrocytes, dystrophic axons and TUNEL-positive cells was performed in cortical layers III to V. Quantitative analysis was done using a morphometric grid within the ocular lens counting 10 microscopic fields of 0.0576 mm per region of interest (normal appearing grey matter and lesion).
This is a morphometric point counting system using a microscope eyepiece graticule and has been suggested as a standard in international consensus reports on quantification of angiogenesis (Vermeulen et al, 1996, 2002).
The area occupied by microvessels was estimated by using a morphometric method of 'point counting' (Ribatti et al, 1995).
The latter number was calculated as determined in the morphometric analysis of podocyte counting methods by Sanden et al [ 10].
Morphometric leukocyte counts for three pre-defined categories of cells (small mononuclear cells, large mononuclear cells and spindle-shaped cells) were generated by two microscopists that individually analyzed four 40x magnification images taken from affected regions of myocardial sections from both cases.
In order to quantitate vascularisation of KAs, MVD was assessed in tumour stroma both by manual count and computer-assisted morphometric analysis of stained areas in at least three regions of interest on each tumour section.
Counts of capillary and arteriolar density per square millimeter were obtained after superimposing a calibrated morphometric grid on each digital image using Adobe Photoshop Software.
