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

CIL:13367*  Cite 
Description

4D image series of HeLa cells cotransfected with CENP-B–GFP (centromeric protein B) and either control (CIL:13368) or Bod1 siRNA (this image). Bod1 is a protein that associates wtih a large macromolecular complex and localizes with kinetochores and spindle poles during mitosis. .

Technical Details

48 h after transfection, cells were split onto 40-mm-diameter glass coverslips (Bioptechs), cultured overnight, and transferred to CO2 independent media with supplements. Cells were maintained at 37°C using an FCS2 chamber in conjunction with an objective heater (Bioptechs). Images were acquired on a restoration microscope (DeltaVision Spectris; Applied Precision) with a 100× 1.35 NA objective and a cooled charge-coupled device camera (CoolSNAP HQ; Roper Scientific). SoftWorx software (Applied Precision) was used for image analysis. Datasets were deconvolved using the constrained iterative algorithm (Swedlow et al., 1997; Wallace et al., 2001) using SoftWorx software. Time courses were presented as maximum intensity projections of deconvolved three-dimentional datasets. Images were loaded into Photoshop (Adobe) or OMERO (http://openmicroscopy.org) and adjusted for display. Image is part of Fig 4B in JCB vol. 179 no. 2 187-197

Biological Sources
NCBI Organism Classification
Homo sapiens
Cell Line
HeLa S3
Cellular Component
chromosome, centromeric region
CENP-B
Biological Context
Biological Process
mitosis
Molecular Function
Bod1 siRNA treatment
Attribution
Names
Iain M. Porter
Sarah E. McClelland
Guennadi A. Khoudoli
Christopher J. Hunter
Jens S. Andersen
Andrew D. McAinsh
J. Julian Blow
Jason R. Swedlow
Published
JCB vol. 179 no. 2 187-197
Pubmed
17938248
Link
Journal of Cell Biology
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL13367
Archival Resource Key (ARK)
ark:/b7295/w9cil13367
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
fluorescence microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
EGFP
Processing History
constrained iterative deconvolution
Sample Preparation
Methods
living tissue
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 237px 0.3312µm
Y 265px 0.3312µm
Z 6px 2.25µm