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

CIL:12020*  Cite 
Description

Mitotic 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 starts at anaphase onset, when sister chromatids separate and sister kinetochores move away from each other as their kinetochore fibers shorten (anaphase A). Almost simultaneously, the two spindle poles move away from each other leading to elongation of the mitotic spindle (anaphase B). The compacted DNA of the chromosome arms excludes the background fluorescence so that chromosome arms appear as dark rod-like structures and the movement of the sister chromatid arms away from each other during anaphase can also be followed. 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. The movie is played at 10 frames per second. Elapsed time is shown in min:sec.

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
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL12020
Archival Resource Key (ARK)
ark:/b7295/w9cil12020
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
Alexa Fluor 488
X-Rhodamine
labeled primary antibody
Processing History
color combine
Data Qualifiers
raw, unprocessed data
suitable for spatial measurements
Sample Preparation
Methods
living tissue
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 260px 0.128µm
Y 198px 0.128µm
Channel Wavelength
1 488,568nm
Time 12 seconds


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