Comparative study of histological and histo-chemical image processing in muscle fiber sections of broiler chicken

Abstract

The image processing of muscular fibers has seen significant improvements over the last decades, from light microscopy acquisition to virtual microscopy and from manual segmentation and fiber typing to automatic. The present study aims to discuss the main tools employed in different histo-enzymological image processing phases applied to muscle fibers, which are respectively: the acquisition, segmentation and fiber typing as well as their efficiency in the determination of morphometric parameters. First, the acquisition: to count the cell numbers within the muscle fiber sections, optical microscopic images with different magnifications (x100, x200, x250, and x400) were compared with virtual slides digitized by Slide Scanner. Second, the segmentation: Three softwares (Fiji “Digitizing pen, Mouse”, Image Pro Plus 10 [semi-automatic], and Cytoinformatics LLC [automatic]) were compared for image segmentation quality and the determining of fiber numbers. Third, manual fiber typing: employing Fiji of segmented images using the 3 previously mentioned softwares were performed to calculate the accuracy of the morphometric parameter (Cross sectional Area [ CSA ], perimeter and Minimal Feret's Diameter [ MFD ]). The results of the acquisition showed that the scanner slides have a better resolution and detected a higher number of cells than when optical microscopes with different magnifications were used; with the latter, the processing can be performed only with an acceptable resolution, where the number of cells is lesser, which unfortunately requires several repetitions and exhausting work. Our findings regarding segmentation indicate that the Cytoinformatics LLC showed the best processing time and the highest quality followed by IP and Fiji. Finally and for the ultimate step, the morphometric parameter calculation: showed that the best accuracy was attained by using Fiji followed by Cytoinformatics and finally by IP. The findings of this study suggest that Fiji (semi-automatic) showed the best quality/price ratio (open access software) for segmentation and fiber typing, but it proved to be time consuming when compared to Image Pro Plus 10 (semi-automatic) and Cytoinformatics LLC (automatic).