Computational protocol: Standardized orthotopic xenografts in zebrafish reveal glioma cell line specific characteristics and tumor cell heterogeneity

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Protocol publication

[…] Individual tumor-bearing or mNSC-transplanted animals were anesthetized using tricaine (160 μg/ml) at 2, 5, 7 and 10 dpt and imaged in a FluoroDish (WPI, Inc., Sarasota, FL, USA) under the spinning disk confocal microscope (Andor). Using MetaMorph software (Molecular Devices, Sunnyvale, CA, USA) and Fiji (), images captured through the zebrafish tumors were reconstructed using a z-stack to analyze tumor volume. Using MetaMorph, a standard cell volume in fluorescent pixels was averaged from six cells. This ‘standard’ cell volume was then applied to the entire z-stack. These measurements were converted into volume (in micrometers cubed) using a MetaMorph software formula that converts a standardized pixel analysis of one ‘cell’ into a volumetric number. Images were analyzed using Fiji/ImageJ and Excel. The same animal was imaged on 2, 5, 7 and 10 dpt, and tumor burden was quantified and plotted for each of these days using Excel. […]

Pipeline specifications

Software tools MetaMorph, ImageJ
Organisms Danio rerio, Homo sapiens/Mus musculus xenograft, Mus musculus, Homo sapiens, Caenorhabditis elegans
Diseases Brain Neoplasms, Death, Glioblastoma, Glioma, Neoplasms