Abstract
A semi-automatic image analysis program, SMART, was used to analyze transmission electron microscopy (TEM) images from four laboratories that participated in an interlaboratory comparison study by Meija et al. on CNC particle size measurement by TEM using conventional manual image analysis approaches. Detailed image-to-image comparisons found that the percentage of “correctly” identified CNCs by SMART was 58% to 78%, while manual was 70% to 87%, depending on TEM image quality from a given laboratory. SMART was able to parameterize image quality, and it was found that SMART had difficulties in CNC identification for images with a combination of higher noise, lower contrast, and higher CNC density. Overall, the SMART image analysis was consistent with the manual approach. SMART showed lower laboratory-laboratory variation as compared to manual, suggesting that the variability of analyst bias of manual approach was removed and demonstrates an opportunity with SMART to improve the standardization of CNC size characterization. An approach to estimate the likelihood of reaching a representative measurement for CNC particle size was developed. SMART area analysis found that less than 10% of CNCs were used in morphology characterization; to assess more CNC material, SMART was used to analyze CNC agglomerates as a proof-of-concept demonstration. The total SMART image analysis time for each laboratory, having between 115 and 244 images, was less than 15 min, after selection of appropriate parameters. The SMART code is now available for the public to use for free at Github™.
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Acknowledgments
The authors would like to thank the USDA Forest Service, Forest Products laboratory for funding this research (Grant Number: 18-JV-11111129-040) and the Renewable Biomaterials Institute at Georgia Institute of Technology. We would like recognize Kai Cui of National Research Council Canada for providing one of the ILC data sets (TEM images, and manual image analysis).
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This work was supported by USDA Forest Service, Forest Products laboratory (Grant Number: 18-JV-11111129-040) and the Renewable Biomaterials Institute at Georgia Institute of Technology.
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The study conception was developed by Sezen Yucel, Robert Moon, Linda Johnston, and Surya Kalidindi. The study design was developed by Sezen Yucel and Robert Moon. Coding design was developed by Sezen Yucel and Surya Kalidindi. Code testing and verification studies was completed by Sezen Yucel and Robert Moon. Results analysis was completed by Robert Moon, Sezen Yucel, and Linda Johnston. TEM images were supplied by Douglas Fox, Byong Chon Park, and E. Johan Foster. First draft of the manuscript was written by Sezen Yucel and Robert Moon. All authors critically reviewed a draft manuscript(s), which resulted in revisions to the document. All authors have read and approved the final manuscript.
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Yucel, S., Moon, R.J., Johnston, L.J. et al. Transmission electron microscopy image analysis effects on cellulose nanocrystal particle size measurements. Cellulose 29, 9035–9053 (2022). https://doi.org/10.1007/s10570-022-04818-w
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DOI: https://doi.org/10.1007/s10570-022-04818-w