Computational protocol: Live imaging of osteoclast inhibition by bisphosphonates in a medaka osteoporosis model

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Protocol publication

[…] For live fluorescence imaging, larvae were anesthetized with 0.01% ethyl 3-aminobenzoate methanesulfonate (tricaine; Sigma A5040) and pictures were taken using a Nikon SMZ1000 stereomicroscope equipped with NIS-Elements BR 3.0 software (Nikon, Japan). For live confocal imaging, larvae were anesthetized with 0.005% tricaine and embedded in 1.5% low-melting-point agarose in a glass-bottomed Petri dish. Confocal pictures were taken with a Zeiss LSM 510 Meta using 405, 488 and 543 nm laser lines for DAPI, GFP and mCherry analysis, respectively. Imaging data were processed using Zeiss LSM Image Browser Version 4.2.0.121, Imaris 7.1.1 (bitplane), Image-Pro Plus 6.0, ImageJ (1.4.3.67) and Adobe Photoshop CS6 (13.0.0.0) software. […]

Pipeline specifications

Software tools NIS-Elements BR, Imaris, ImageJ
Application Microscopic phenotype analysis
Organisms Oryzias latipes, Homo sapiens
Diseases Bone Diseases, Osteoporosis
Chemicals Diphosphonates, Etidronic Acid, Alendronate