Mitotic cells were observed using a Leica DMI6000 (Leica Microsystems, Germany) Microscope and images were acquired with a Hamamatsu FLASH4.0 (Hamamatsu, Japan) camera, using the HCX PL APO CS 63x/1.30 GLY 21°C objective, and the LAS X Software. Images were taken in Z-Stacks in a range of 10-14μm, with a distance between planes of 0.2μm.
We consider as centriole amplification, when mitotic cells presented more than four (>4) centrioles. In order to obtain the percentage of cells with extra centrioles, at least 100 cells were analyzed for centriole number per condition and per experiment and only centrioles positive for the two centriolar markers (Centrin-1 and CP110) were considered. Centrosomes were quantified manually, using the Fiji/Image J Software (National Institutes of Health).
We consider as centriole amplification, when mitotic cells presented more than four (>4) centrioles. In order to obtain the percentage of cells with extra centrioles, at least 100 cells were analyzed for centriole number per condition and per experiment and only centrioles positive for the two centriolar markers (Centrin-1 and CP110) were considered. Centrosomes were quantified manually, using the Fiji/Image J Software (National Institutes of Health).
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