Computational protocol: Neurotrophically Induced Mesenchymal Progenitor Cells Derived from Induced Pluripotent Stem Cells Enhance Neuritogenesis via Neurotrophin and Cytokine Production

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Protocol publication

[…] Fluorescent DRG image tiles were processed with MetaMorph (Molecular Devices, Sunnyvale, CA), and digitally stitched together using open source software Fiji/NIH ImageJ . All grayscale images were thresholded to create binary bitmap images. Neurite extension densities were measured using the Sholl analysis plugin function within Fiji. The algorithm automatically retrieves data from two‐dimensional (2D) or three‐dimensional (3D) images to perform a regression analysis and generate the metrics for Sholl‐based dendrite arborization . All images were taken at ×4 magnification using an inverted microscope (Olympus IX81) with a motorized stage controlled through MetaMorph, and the scale used was 620 px = 1 mm.Neurite extension lengths were quantified based on images of DRGs cultured on 2D aligned nanofibrous poly‐ɛ‐caprolactone (PCL) scaffolds prepared by electrospinning (see below). All images for scaffold cultures were taken at ×10 using the inverted Olympus scope (IX81). Tracings were converted from pixel length into μm measurements using the scale of 1245 px = 800 μm. […]

Pipeline specifications

Software tools MetaMorph, ImageJ, Sholl analysis
Application Microscopic phenotype analysis
Organisms Gallus gallus