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

CIL:8078*  Cite 
Description

Metaphase PtK1 cell microinjected with X-rhodamine-labeled tubulin and Alexa 488-labeled CENP-F antibodies, to fluorescently label kinetochore fibers (red) and kinetochores/spindle poles (green), respectively. The movie shows that sister kinetochore pairs oscillate between the two spindle poles as their respective kinetochore fibers lengthen and shorten. The top kinetochore of the first sister pair on the right exhibits abnormal attachment to spindle microtubules, as it is bound to microtubules coming from both spindle poles, rather than just one (merotelic kinetochore orientation). As a consequence of this mis-attachment, the merotelically oriented kinetochore remains relatively stationary while its normally oriented sister oscillates between poleward and anti-poleward movements. Digital images were collected with an Orca ER cooled CCD camera (Hamamatsu Photonics) coupled to a Yokogawa spinning disk confocal CS10 unit (Perkin Elmer Life Sciences Wallac), which was attached to a Nikon TE300 inverted microscope. Illumination was achieved via an Argon/Krypton laser and 488 nm and 568 nm filters in an excitation filter wheel (Sutter Instruments) through a 60 x 1.4 NA Plan-Apochromatic DIC objective lens. Nearly simultaneous fluorescence images were acquired at 488 and 568 nm at a single focal plane every 20-60 seconds. The movie is played at 10 frames per second. Elapsed time is shown in min:sec. Corresponds to video 1 in Curr Biol. 2004 Dec 14;14(23):2149-55.

Biological Sources
NCBI Organism Classification
Potorous tridactylus
Cell Type
epithelial cell
Cell Line
Pt K1 (NBL-3)
Cellular Component
kinetochore
spindle microtubule
Biological Context
Biological Process
attachment of spindle microtubules to kinetochore
mitosis
Attribution
Names
Daniela Cimini
Lisa A Cameron
Edward D Salmon
Published
Curr Biol. 2004 Dec 14;14(23):2149-55.
Pubmed
15589159
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL8078
Archival Resource Key (ARK)
ark:/b7295/w9cil8078
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
7
Visualization Methods
Alexa Fluor 488
X-Rhodamine
labeled primary antibody
Processing History
color combine
Data Qualifiers
raw, unprocessed data
Sample Preparation
Methods
living tissue
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 302px 0.128µm
Y 295px 0.128µm
Channel Wavelength
1 488,568nm
Time 20 seconds