DiameterJ: A validated open source nanofiber diameter measurement tool

Hotaling, Nathan; Bharti, Kapil; Kriel, Haydn; Simon, Carl G.

doi:10.1016/j.biomaterials.2015.05.015

Cited by 440 publications

(387 citation statements)

References 53 publications

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“…Samples were mounted on aluminum stubs using carbon tape and were sputter coated with platinum using Leica EM ACE600. The average diameter and pore size of the fibrous scaffolds were analyzed using DiameterJ, 47 and ImageJ 48 software, respectively.…”

Section: Methodsmentioning

confidence: 99%

Regulation of Epithelial-to-Mesenchymal Transition Using Biomimetic Fibrous Scaffolds

Ravikrishnan¹,

Özdemir²,

Bah³

et al. 2016

ACS Appl. Mater. Interfaces

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Epithelial-to-mesenchymal transition (EMT) is a well-studied biological process that takes place during embryogenesis, carcinogenesis and tissue fibrosis. During EMT, the polarized epithelial cells with a cuboidal architecture adopt an elongated fibroblast-like morphology. This process is accompanied by the expression of many EMT-specific molecular markers. While the molecular mechanism leading to EMT has been well established, the effects of matrix topography and microstructure have not been clearly elucidated. Synthetic scaffolds mimicking the mesh-like structure of the basement membrane with an average fiber diameter of 0.5 μm and 5 μm were produced via electrospinning of poly(ε-caprolactone) (PCL) and were used to test the significance of fiber diameter on EMT. Cell-adhesive peptide motifs were conjugated to the fiber surface to facilitate cell attachment. Madin-Darby Canine Kidney (MDCK) cells grown on these substrates showed distinct phenotypes. On 0.5 μm substrates, cells grew as compact colonies with an epithelial phenotype. On 5 μm scaffolds, cells were more individually dispersed and appeared more fibroblastic. Upon addition of hepatocyte growth factor (HGF), an EMT inducer, cells grown on the 0.5 μm scaffold underwent pronounced scattering, as evidenced by the alteration of cell morphology, localization of focal adhesion complex, weakening of cell-cell adhesion, and upregulation of mesenchymal markers. By contrast, HGF did not induce a pronounced scattering of MDCK cells cultured on the 5.0 μm scaffold. Collectively, our results show that the alteration of the fiber diameter of proteins found in the basement membrane may create enough disturbances in epithelial organization and scattering that might have important implications in disease progression.

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Section: Methodsmentioning

confidence: 99%

Regulation of Epithelial-to-Mesenchymal Transition Using Biomimetic Fibrous Scaffolds

Ravikrishnan¹,

Özdemir²,

Bah³

et al. 2016

ACS Appl. Mater. Interfaces

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“…Small gauge (48 and 50 gauge) steel wire was chosen as an ideal material for training because 1) it has a highly uniform diameter and 2) because the diameter is large enough that it can be verified by other measurements [23]. Specific sizes of wire were chosen because they were commercially available and because they were small enough to be imaged using the “high magnification” settings within the SEM yet large enough to be sized by alternative means (calipers, light microscopy, impedance).…”

Section: Methodsmentioning

confidence: 99%

“…From SEM micrographs the most prevalent measurement made to assess nanofiber morphology is the fiber diameter. [23] Currently, manual measurement of fiber diameter from fiber SEM images is done using line tools in image analysis programs such as ImageJ/FIJI [24,25]. Recently, a publication comparing software tools and manual measurements on images of nanofibers showed that hundreds to thousands of nanofiber measures were required to converge on a consistent result.…”

Section: Introductionmentioning

confidence: 99%

“…To help improve the process of analyzing micrographs for assessing fiber morphology, several algorithms have been published and recently, DiameterJ, an open access tool for assessing fiber morphology via a plugin for ImageJ, was developed [23,26]. DiameterJ enables automated analysis of scanning electron micrographs to rapidly determine fiber diameter, pore size, porosity, fiber intersection density and the characteristic lengths of fibers between intersections.…”

Section: Introductionmentioning

confidence: 99%

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Training to Improve Precision and Accuracy in the Measurement of Fiber Morphology

et al. 2016

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An estimated $7.1 billion dollars a year is spent due to irreproducibility in pre-clinical data from errors in data analysis and reporting. Therefore, developing tools to improve measurement comparability is paramount. Recently, an open source tool, DiameterJ, has been deployed for the automated analysis of scanning electron micrographs of fibrous scaffolds designed for tissue engineering applications. DiameterJ performs hundreds to thousands of scaffold fiber diameter measurements from a single micrograph within a few seconds, along with a variety of other scaffold morphological features, which enables a more rigorous and thorough assessment of scaffold properties. Herein, an online, publicly available training module is introduced for educating DiameterJ users on how to effectively analyze scanning electron micrographs of fibers and the large volume of data that a DiameterJ analysis yields. The end goal of this training was to improve user data analysis and reporting to enhance reproducibility of analysis of nanofiber scaffolds. User performance was assessed before and after training to evaluate the effectiveness of the training modules. Users were asked to use DiameterJ to analyze reference micrographs of fibers that had known diameters. The results showed that training improved the accuracy and precision of measurements of fiber diameter in scanning electron micrographs. Training also improved the precision of measurements of pore area, porosity, intersection density, and characteristic fiber length between fiber intersections. These results demonstrate that the DiameterJ training module improves precision and accuracy in fiber morphology measurements, which will lead to enhanced data comparability.

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“…20 Kurtosis for angle of orientation graphs was measured in Excel. Kurtosis is a 6 measure of "tailedness" of the probability distribution of a real-valued random variable.…”

Section: Scaffold Characterizationmentioning

confidence: 99%

Improving myoblast differentiation on electrospun poly(ε‐caprolactone) scaffolds

Abarzúa-Illanes

Padilla

Ramos

et al. 2017

J Biomedical Materials Res

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Polymer scaffolds are used as an alternative to support tissue regeneration when it does not occur on its own. Cell response on polymer scaffolds is determined by factors such as polymer composition, topology, and the presence of other molecules. We evaluated the cellular response of murine skeletal muscle myoblasts on aligned or unaligned fibers obtained by electrospinning poly(ε-caprolactone) (PCL), and blends with poly(lactic-co-glycolic acid) (PLGA) or decorin, a proteoglycan known to regulate myogenesis. The results showed that aligned PCL fibers with higher content of PLGA promote cell growth and improve the quality of differentiation with PLGA scaffolds having the highest confluence at over 68% of coverage per field of view for myoblasts and more than 7% of coverage for myotubes. At the same time, the addition of decorin greatly improves the quantity and quality of differentiated cells in terms of cell fusion, myotube length and thickness, being 71, 10, and 51% greater than without the protein, respectively. Interestingly, our results suggest that at certain concentrations, the effect of decorin on myoblast differentiation exceeds the topological effect of fiber alignment. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2241-2251, 2017.

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DiameterJ: A validated open source nanofiber diameter measurement tool

Cited by 440 publications

References 53 publications

Regulation of Epithelial-to-Mesenchymal Transition Using Biomimetic Fibrous Scaffolds

Regulation of Epithelial-to-Mesenchymal Transition Using Biomimetic Fibrous Scaffolds

Training to Improve Precision and Accuracy in the Measurement of Fiber Morphology

Improving myoblast differentiation on electrospun poly(ε‐caprolactone) scaffolds

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