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Adopting automated image analysis tool for fibrin network: Can we obtain clot properties for practical application?

Sir, Fibrin clots are formed through activation of the coagulation pathway, resulting in the formation and lateral aggregation of fibrin fibers. This threedimensional architecture of a clot is closely related to clot stability, and various pathologic conditions are known to be associated with an altered clot structure.1-3 However, most of the traditional coagulation tests measure clotting times and evaluate the activity of coagulation cascade. Because direct structural evaluation for fibrin network requires high magnification, it could have been mainly achieved by electron microscopy and carried out in the field of research. Although fibrin network analysis offers insight into clot formation and dissolution, the practical use has been seldom attempted.4 Because sample preparation for scanning electron microscopy (SEM) was time consuming and the analysis was difficult, it has not been considered for practical assay. Yet, technical development and automation of SEM, recently, have potentiated the application of fibrin network analysis in clinical laboratories. Several parameters including fibrin diameter, branching points, and porosity are able to be analyzed in SEM images of fibrin network. These parameters provide a basic approach for quantitating fibrin network. However, there have been no, so far as we are aware, guideline and discussions on an analysis process; the gold standard for assessing clot images has been manual measurement. A standardized guideline is required for the analysis process; therefore, the measuring process could be automated. Recently, some studies have used an automated analyzer for fibrin network analysis; however, the system not only requires a specific SEM but also expenses cost.5 Here, we adopted a free open source tool—DiameterJ, which has been used for nanofiber diameter measurement—to measure fibrin network properties.6 In this study, we explored the applicability of automated image analysis tool for fibrin network structure. Additionally, we investigated the possibilities of practical application of fibrin network analysis and discussed the requirements of the image analysis process to establish an appropriate analysis guideline.