Comparative Study of Histological and Histochemical Image Processing in Muscle Fibre Sections of Broiler Chicken

Abstract Image processing of muscular fibre has significantly improved over the last decades, from light microscopy acquisition to virtual microscopy and from manual segmentation and fibre typing to automatic. The study aims to discuss main tools used in different histoenzymological image processing ‎phases of muscle fibre, which are consecutively, acquisition, segmentation and fibre typing and theirs ‎efficacy in morphometric parameters’ determination. Firstly, the acquisition, optical microscopic image with different magnifications ‎‎(x100, x200, x250 and x400) were compared with virtual slides digitized by Slide Scanner, in ‎muscle fibre sections, for cell number counting. Secondly, the segmentation, three software ‎‎(Fiji «Digitizing pen, Mouse», Image Pro Plus 10 -semi-automatic- and Cytoinformatics LLC -automatic- ) were compared ‎for image segmentation quality and fibbers number determination. Thirdly, manual fibre typing with Fiji of segmented images using three software cited previously were performed to calculate the accuracy of morphometric parameter (CSA, perimeter and DMF). Results of acquisition showed that scanner slide have ‎a better resolution and detected a higher number of cells compared to optical microscopic with ‎different magnifications. For this latter, the processing can be performed only with an ‎acceptable resolution, where the number of cells is lesser, which requires unfortunately several ‎repetitions and exhausting work. Our findings regarding segmentation indicate that ‎Cytoinformatics LLC showed the best processing time and the highest quality followed by IP ‎and Fiji. For the last step, morphometric parameter calculation showed the best accuracy 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 fibre typing, but time consuming compared to ‎ Image Pro Plus 10 (semi-automatic) and Cytoinformatics LLC‎ (automatic) which are a paid service.