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Description

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.

Technical Details

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.

Biological Sources

NCBI Organism Classification
Homo sapiens
(human)
Cell Type
hepatocyte
Cell Line
HuH-7
Cellular Component
chromosome
spindle microtubule

Biological Context

Biological Process
mitotic metaphase
cytokinesis
chromosome segregation
chromosome organization
Human Development Anatomy
liver
Human Disease
sclerosing hepatic carcinoma

Attribution

Names
Michael Cammer
Phyllis Novikoff
Date
12/1993

Grouping

This image is representative of a group of similar images.

Imaging

Image Type
recorded image
Imaging Mode
single-spot confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
distribution of nucleic acids
Visualization Methods
propidium iodide
Fluorescein (FITC)
Processing History
volume reconstruction
Data Qualifiers
processed data

Sample Preparation

Methods
glutaraldehyde fixed tissue
formaldehyde fixed tissue
Relation To Intact Cell
dispersed cells in vitro

Dimensions

Spatial Axis Image Size Pixel Size
X 2752px ——
Y 2752px ——
*CIL – Cell Image Library accession number. Please use this to reference an image.