Computational protocol: Distribution of proteins within different compartments of tendon varies according to tendon type

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Protocol publication

[…] To assess the distribution of specific proteins, two independent assessors (K.J.K. and J.N.), blinded to tendon type and staining condition, graded the staining intensity of the immunohistochemical images. In each image, the IFM and FM were graded separately on a scale from 0 to 5, with 0 representing no staining and five representing very intense staining. Any images with patchy staining or cutting artefacts were not included in the final analysis. Inter‐observer variability was assessed by calculating linear weighted Kappa statistics (Viera & Garrett, ) using an online software tool (http://vassarstats.net/kappa.html). [...] Elastic fibres were sparsely distributed throughout the IFM and FM, so elastin content was assessed by measuring the length of each fibre and expressing total fibre length relative to IFM or FM area (imagej Software, NIH, USA). Elastic fibre thickness appeared constant across all images and so was approximated to one pixel for all fibres. […]

Pipeline specifications

Software tools VassarStats, ImageJ
Applications Miscellaneous, Microscopic phenotype analysis
Diseases Tendinopathy