Behavior of a patient-derived human fibroblast line (PD-NC4) containing a mutated chromosome 4 in which the normal centromere is epigenetically silenced and a neo-centromere has formed 25 Mbp down the q arm at a site that lacks centromere repeats. Shown is a z-series of the PD-NC4 chromosome from a metaphase spread stained for CENP-B (red), centromere (ACA, green) and DNA (DAPI blue). See Fig 1D in Bassett et al. 2010. Epigenetic centromere specification directs aurora B accumulation but is insufficient to efficiently correct mitotic errors. J Cell Biol 190:177-185.
Cells were metaphase-arrested with nocodazole, swollen in 75 mM KCl, centrifuged onto glass slides, and fixed with formaldehyde before processing for indirect immunofluorescence. Digital z-series images were captured with an ORCA AG CCD mounted on DMI6000B Leica microscope using a 100X 1.4 NA objective lens and subsequently processed by deconvolution.
| Spatial Axis | Image Size | Pixel Size |
|---|---|---|
| X | 101px | —— |
| Y | 91px | —— |
| Z | 20px | —— |
