Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: Intravital Imaging of Tumor Cell Motility in the Tumor Microenvironment Context

doi: 10.1007/978-1-4939-7701-7_14

Figure Lengend Snippet: Intravital Systems Microscopy: Multiparametric SVM classification of tumor cell locomotion and microenvironmental parameters (a) Identifying and quantifying fast-locomoting cells. Left panels, raw images of an individual z-section from the 4D stack (at 0 and 60 min; 0′ and 60′). Middle panels, 0′ was subtracted from 60′, resulting in Δ60 (top); image was then thresholded/binarized (bottom). Right panels, results of motility analysis including quantification of fast locomoting cells (top) and the overlay with the 0′ image (bottom). Scale bar 50 μm. (b) Identifying and quantifying invadopodia in slow locomoting cells. Raw images of a cell at time 0, with fully extended invadopodium at 3 min and partially retracted invadopodium at 15 min. Overlays Δ3 and Δ15 show invadopodia extension highlighted in magenta. Scale bar 10 μm. (c) Binarized images used for extraction of microenvironmental parameters. Image in Fig. 1c was separated and thresholded, resulting in binary images of collagen (magenta), tumor cells (green), macrophages (cyan) and blood vessels (red). (c′) Collagen fiber map (left) and dimensionless straightness histogram (right) from ctFIRE software. (d) 3D projection of SVM classification results. Red spheres denote slow locomotion, blue-fast locomotion, green- misclassifications. The size of the spheres indicates the number of locomoting cells in the FOV. Dmax (μm) stands for the diameter of the largest blood vessel in the FOV, macrophages (%) and collagen (%) stand for the thresholded area in respective channels

Article Snippet: The ctFIRE software requires MATLAB compiler runtime (MCR 7.17 2012a) installation.

Techniques: Microscopy, Extraction, Software