Surface representation of tripolar mitosis in a liver cancer cell line. Normally, dividing cells form a single metaphase plate of paired chromosomes which are pulled apart by opposing microtubule spindles. This image shows abnormal division by three sets of spindles (cyan or light blue) attached to mis-segregated chromosomes (yellow). Other images in this group show the original fluorescence images.
HuH-7 cells are an immortalized cultured cell line derived from hepatic carcinoma. Cells were grown on 1.5 glass coverslips, fixed with formaldehyde and glutareldehyde, extracted, and labeled for fluorescence microscopy with commercial antibodies to alpha- or beta-tubulin and propidium iodide. The coverslip was mounted on a slide in PBS and glycerol with n-propyl gallate as an antifading agent. Imaging was performed with a BioRad MRC 600 laser scanning confocal microscope with a Kr/Ar laser with lines at 488 and 568 nm. The microscope was a fixed tube length Nikon Diaphot with 60X N.A. 1.4 phase 3 optics. Z series was collected at less than 0.5 um step sizes. The raw data were filtered with custom C++ routines on a Silicon Graphics computer and then volume reconstruction performed with VoxelView software that treated the structures as solids with light projected onto them. The original spatial scale has been lost. The original confocal scans were 512 X 512 pixels in XY with each pixel oversampled. Subsequent processing in PhotoShop lost the original scale. The diameter of this cell is approximately 20 um.
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