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Validated Open Source Nanofiber Diameter Measurement Tool



Nathan A. Hotaling, Kapil Bharti, Haydn Kriel, Carl G. Simon Jr.


Despite the growing use of nanofiber scaffolds for tissue engineering applications, there is not a validated, readily available (commercial or open source) solution for rapid, automated analysis of nanofiber diameter from scanning electron microscope (SEM) micrographs. Thus, the goal of this study was to create a user friendly ImageJ/FIJI plugin that would analyze SEM micrographs of nanofibers to determine nanofiber diameter on a desktop computer within 60 seconds. Additional design goals included 1) compatibility with a variety of existing segmentation algorithms and 2) an open source code to enable further improvement of the plugin. Using existing algorithms for centerline determination, Euclidean distance transforms and a novel pixel transformation technique, a plugin called “DiameterJ” was created for ImageJ/FIJI. The plugin was validated using 1) calibration images of white lines on a black background and 2) SEM images of steel reference wires of known diameters. DiameterJ analyzed SEM micrographs in 20 seconds, was accurate within 2% of known values, was more than 10-times more accurate than other open source software, provided hundreds of times the sampling of manual measurement, and was hundreds of times faster than manual assessment of nanofiber diameter. DiameterJ enables users to rapidly and thoroughly determine the properties of nanofiber scaffolds, potentially allows new insights to be formed into fiber diameter distribution and cell response, and may be useful for determining lot release criteria for nanofiber scaffolds used in regenerative medicine and other applications.


nanofiber, ImageJ, fiber diameter


Hotaling, N. , , K. , Kriel, H. and Simon, C. (2015), Validated Open Source Nanofiber Diameter Measurement Tool, Biomaterials (Accessed April 22, 2024)
Created April 30, 2015, Updated February 19, 2017