Automated Fiber Diameter and Porosity Measurements of Plasma Clots in Scanning Electron Microscopy Images
, Stephen R. Baker 1,4, Zelda de Lange-Loots 2,3, Aleksander Siniarski 5,6, Rustem I. Litvinov 7, Caroline S. B. Veen 8, Moniek P. M. de Maat 8, John W. Weisel 7, Robert A. S. Ariëns 4 and Martin Guthold 1,*Round 1
Reviewer 1 Report
Highly detailed analysis which will be of significant use to the community in using the appropriate DiameterJ algorithms. The analysis also points to the limitations of SEM appropriately.
I have two questions; (1) when reanalyzing the data set 2 at higher magnification, was the comparison with manual method made with manual measurements made at higher magnification images, i.e., consistent comparisons? (2) The comparisons are made with manual measurements to benchmark the algorithms. How valid is this comparison which assumes manual measurements are the gold standard?
Author Response
Please see the attachment
Reviewer 2 Report
The manuscript of Ali Darai et al. is dedicated to the study of fibrin morphology using scanning electron microscopy. The fibrin structure plays a vital role in maintaining hemostasis. Mass/length ratio of fiber, network porosity, branching degree (branch point density) are important structural characteristics of fibrin network, which determine its rheological and biochemical properties, including resistance of fibrin to plasmin hydrolysis. A manual scanning electron microscopy method has been used in many studies. However, to evaluate the advantage of a reliable automated method for structural analysis of fibrin network images to measure the above characteristics the authors have carried out comprehensive and detailed studies of the structure of fibrin clots formed from the plasma of healthy people and patients suffering from various hemostasis disorders.
All studies have been performed extremely thoroughly. As shown by numerous studies, in pathology, including oxidative stress, finely porous fibrin is formed, which has an increased resistance to plasmin hydrolysis. Clots formed from thicker fibers generally have looser and less stiff networks that are more permeable and more susceptible to fibrinolysis. Therefore, one can fully support the authors of this manuscript that an automated analysis method would also be useful in the development of standardization protocols to investigate the structure of blood clots in hemostasis and thrombosis. The manuscript is well structured, and all results are statistically significant and reproducible.
Without a doubt, the manuscript may be recommended for publication. I have no remarks.
Author Response
Please see the attachment.
