Computational protocol: Migratory and adhesive properties of Xenopus laevis primordial germ cells in vitro

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Protocol publication

[…] 30 mm Petri dishes were incubated with 2 ml of 5% (w/v) bovine serum albumin (BSA or Albumin Fraction V, Carl Roth GmbH, Karlsruhe, Germany) in 1× PBS overnight, washed three times with 1× PBS and then covered with agarose (0.5% (w/v) in 0.8× MBSH). After the gel polymerized, 0.8× MBSH was added on top to cover the agarose gel. Cells, isolated as described above, were transferred with a pipette under the agarose layer, between the BSA-covered bottom of the Petri dish and the agarose gel. Cells were monitored by time-lapse imaging using a LumarV.12 fluorescence stereomicroscope (Zeiss, Jena, Germany) and the AxioPlan software (Zeiss, Jena, Germany). Tracking was made using Fiji MTrackJ plug-in and analyzed using ImageJ Chemotaxis Tool plug-in. […]

Pipeline specifications

Software tools MTrackJ, ImageJ
Applications Laser scanning microscopy, Microscopic phenotype analysis
Organisms Xenopus laevis, Mus musculus, Danio rerio, Drosophila melanogaster
Diseases Neoplasms, Germ Cell and Embryonal