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

CIL:28777*  Cite 
Description

Spinning disk confocal time-lapse imaging of the first mitosis in a C. elegans embryo that has been depleted of KNL-2 by RNAi. GFP-histone and GFP-γ-tubulin were imaged (strain TH32). Pronuclear migration is slightly disrupted by a chromatin bridge resulting from failed meiotic segregation of the maternal chromatin. As seen in CeCENP-A depletion (CIL 28776), chromosome condensation is disrupted, chromosomes fail to form a proper metaphase plate, and the spindle poles move away from the chromatin masses prematurely. Anaphase segregation fails, and chromatin masses are left in the middle of the cell. Also note the meiotic defect that results in a defective oocyte pronucleus (left side of image at the beginning of the videos; this is the embryo anterior). The video is ∼16 min long, and the frame is ∼30 µm top to bottom. Images were acquired at 10-s intervals.

Technical Details

Images were acquired via a spinning disk confocal microscope (CSU10; McBain Instruments) mounted on an inverted microscope (TE2000e; Nikon). Strain TH32 coexpressing GFP-histone H2b and GFP–γ-tubulin was imaged using a 60× 1.4 NA plan Apo objective with 1.5× auxiliary magnification and a cooled CCD camera (Orca ER; Hamamatsu) binning 2 × 2. Strain OD31 expressing GFP–KNL-2 was imaged in the same manner without the 1.5× auxiliary magnification.

Biological Sources
NCBI Organism Classification
Caenorhabditis elegans
Cell Type
embryo
Cellular Component
nuclear chromosome
gamma tubulin
spindle pole
chromatin bridge
Biological Context
Biological Process
KNL-2 depletion
mitosis
Attribution
Names
Paul S. Maddox
Francie Hyndman
Joost Monen
Karen Oegema
Arshad Desai
Published
J Cell Biol. 2007 Mar 12;176(6):757-63. Epub 2007 Mar 5.
Pubmed
17339379
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL28777
Archival Resource Key (ARK)
ark:/b7295/w9cil28777
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
spinning disk confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
EGFP
Processing History
unprocessed raw data
Sample Preparation
Methods
living tissue
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 450px ——
Y 288px ——
Time 10 seconds 99