Comparison of threshold algorithms for automatic processing of fat crystal microscopic images based on ImageJ

Abstract Microscopic image analysis is a crucial tool in fat crystallization research, enabling the analysis of crystal size, network structure, fractal dimension and other parameters through binarization. It is essential to seek an appropriate thresholding algorithm to binarize fat crystal images, which plays a vital role in image segmentation. In this article, the effectiveness of 17 thresholding algorithms such as Default, Mean, IsoData, Otsu, Li and Triangle were analyzed in processing fat crystal images with different shapes, background colors and image intensities. This was expected to discover a stable and objective thresholding algorithm for the binarization of fat crystal images. The performance evaluation was conducted according to the peak signal noise ratio (PSNR), structural similarity index (SSIM) and region non‐uniformity (RNU) parameter. Moreover, the comparative analysis of crystal size error, crystal area fraction and intraclass correlation coefficients (ICC) for fractal dimension values would provide a foundation for the selection of thresholding techniques for fat crystal network images. The results indicated that the Default algorithm exhibited remarkable robustness and applicability with high‐quality and stable outputs in fat crystal image processing.