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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:8058*  Cite 
Description

This HeLa cell, which is expressing both a kinetochore marker (EGFP-CENP-B) and a centrosome marker (Venus-centrin), has been depleted of the spindle assembly checkpoint protein Mad2. In the absence of Mad2, the cell rapidly progresses from early prometaphase through anaphase and cytokinesis. The spindle assembly checkpoint functions to generate a WAIT signal until all sister kinetochore pairs become properly attached to the mitotic spindle. Loss of checkpoint function leads to anaphase initiation before kinetochores are completely aligned at the spindle equator. Despite the rapid nature of these divisions, the majority of the kinetochores appear to separate properly in Mad2-depleted cells. However, at least a few lagging chromosomes are frequently observed in these cells, demonstrating the importance of checkpoint function. Note that the frame width is approximately 30 microns. This movie is part of a group that captured first place at the 2007 ASCB Celldance contest.

Technical Details

This HeLa cell was analyzed by time-lapse microscopy 48 hours after DNA transfection and 36 hours after transfection with 120 nM of Mad2-specific siRNAs (GGAUGACAUGAGGAAAAUA and GCGUGGCAUAUAUCCAUCU). Prior to filming the cell was switched to CO2-independent media (Life Technologies). The cell was imaged with a Deltavision RT system (Applied Precision) equipped with a CCD camera, 60X 1.42 NA lens (Olympus) and a 37C environmental chamber (Applied Precision). Z-stacks containing five focal planes with 0.5 um spacing were acquired at intervals of 5 seconds. Images were deconvolved and projected as a 2-D image stack using Softworx software (Applied Precision). The movie was cropped, bleach corrected, and contrast adjusted using Image J. Display rate is 20 frames per second. Original resource created on February 18, 2007.Original resource: Deltavision Image stack

Biological Sources
NCBI Organism Classification
Homo sapiens
Cell Type
cervical cancer
epithelial cell
Cell Line
HeLa
Cellular Component
kinetochore
spindle pole centrosome
Attribution
Names
Jason Stumpff
George von Dassow
Michael Wagenbach
Charles Asbury
Linda Wordeman
Published
Dev Cell. Feb;14(2):252-62. (2008)
Pubmed
18267093
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL8058
Archival Resource Key (ARK)
ark:/b7295/w9cil8058
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
time lapse microscopy
widefield illumination
fluorescence microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
VenusFP
EGFP
Processing History
3D-deconvolution
Data Qualifiers
processed data
Sample Preparation
Methods
living tissue
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 290px ——
Y 238px ——
Time 5 seconds 200