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Quantification of equine dermal collagen by two morphometric techniques: point counting and color segmentation

ABSTRACT

The types I and III collagens present different tonalities of birefringence in histological sections stained with Picrosirius red, that can be analyzed under a polarized light microscope. Based on this property, collagens can be quantified by histomorphometry. However, many variables can affect the color distribution in the histological image, and the appropriate choice of the analysis parameters have a significant influence on the final result. The objective of this study was to compare the histomorphometric quantification of collagen in the equine skin by counting points planimetry and color segmentation with different configurations to determine the best method of evaluation. For planimetry, three different graticules (391, 588 and 792 intersections) were used and, for color segmentation, six combinations of hue and brightness in ImageJ software. The values were submitted to the Friedman test followed by Tukey with 5% significance. The results showed that the quantification of collagens in the graticule of 792 intersections was equivalent to the results of color segmentation with a brightness of 1-255 and hue of 0-42 and 43-120 for collagens type I and III, respectively. Automatic analysis of the color segmentation, using suitable configuration for brightness and hue can replace the counting points planimetry reliably and safely.

Keywords:
image analysis; histomorphometry; skin

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