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

CIL:11988*  Cite 
Description

A comparison of kinesin-6 anaphase dynamics in a cell with a bipolar or monopolar spindle. (left) TIRF time lapse of kinesin-6 (Pav-GFP; green) and mCherry-tubulin (red) in a wild-type S2 cell with a bipolar spindle from metaphase to anaphase. (right) TIRF time lapse of kinesin-6 (Pav-GFP; green) and mCherry-tubulin (red) in a cell with a monopolar spindle (S2 cells treated with dsRNA to Klp61F (kinesin 5 motor) and BubR1 (checkpoint protein)). This video corresponds to the image in Fig. 5 B and video 5 from J Cell Bio 186:727-738, 2009. Total internal reflection microscopy was performed using a microscope (Perfect-focus TE2000; Nikon) with a 100×, 1.45 NA objective (Nikon) and illumination from either a 488 nm argon laser (100 mW) or a 491 nm solid-state laser (100 mW) and a 561-nm solid-state laser (50 mW). For dual-color TIRF microscopy, we used a triplepass dichroic filter (z491/561/633rpc) and changed the emission filter (ET525/50 or ET595/50; both from Chroma Technology Corp.) with a filter wheel placed before the camera. In some cases, an excitation notch filter was used in the filter cube (NF01-405/488/561/635; Semrock, Inc.). Images were typically captured every 2-3 s with a 50-200 ms exposure with an EM charge-coupled device camera (iXon; Andor Technology). The microscope was controlled and images were acquired using open source MicroManager software (http://www.micro-manager.org).

Biological Sources
NCBI Organism Classification
Drosophila melanogaster
Cell Type
epithelial cell
Cell Line
S2
Cellular Component
kinesin-6
spindle
Biological Context
Biological Process
mitotic anaphase
mitotic metaphase
bipolar spindle
monopolar spindle
kinesin 5 knockdown
BubR1 knockdown
Attribution
Names
Ronald D. Vale
James A. Spudich
Eric R. Griffis
Published
J Cell Bio 186:727-738, 2009
Pubmed
19720876
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL11988
Archival Resource Key (ARK)
ark:/b7295/w9cil11988
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
TIRF
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
EGFP
mCherryFP
Processing History
unprocessed raw data
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 624px 0.0924µm
Y 302px 0.0924µm
Channel Wavelength
1 488, 561nm


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