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Description

Spinning disk confocal time-lapse imaging of the first mitosis in a C. elegans embryo expressing GFP-KNL-2 (strain OD31) in which KNL-1 has been depleted by RNAi. Discrete foci of GFP signal within the nucleus coalesce along curvilinear chromosomes during prophase. After nuclear envelope breakdown, chromosomes fail to congress to the metaphase plate, but GFP-KNL-2 localization is evident. As seen after CeCENP-A depletion and CeCENP-C depletion (CIL 28779 and CIL 28782, respectively), foci of GFP-KNL-2 formed along the spindle after anaphase onset. The video is ?11 min long, and the frame is ?30 µm top to bottom. Images were acquired at 10-s intervals (see Materials and methods), and corresponding data are shown in Fig. 3 in J Cell Biol. 2007 Mar 12;176(6):757-63. Epub 2007 Mar 5.

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 GFPâ��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 GFPâ��KNL-2 was imaged in the same manner without the 1.5× auxiliary magnification.

Biological Sources

NCBI Organism Classification
Caenorhabditis elegans
(Nematode)
Cell Type
embryo
Cellular Component
nuclear chromosome
nuclear chromosome
spindle

Biological Context

Biological Process
KNL-1 depeletion
mitosis
mitotic nuclear envelope disassembly
prophase
anaphase

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

Grouping

This image is part of a group.

Imaging

Image Type
recorded image
Imaging 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 262px ——
Y 153px ——
Time 10 sec 70
*CIL – Cell Image Library accession number. Please use this to reference an image.