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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 (

Biological Sources

NCBI Organism Classification
Drosophila melanogaster
(Common fruit fly)
Cell Type
epithelial cell
Cell Line
Cellular Component

Biological Context

Biological Process
mitotic anaphase
mitotic metaphase
bipolar spindle
monopolar spindle
kinesin 5 knockdown
BubR1 knockdown


Ronald D. Vale
James A. Spudich
Eric R. Griffis
J Cell Bio 186:727-738, 2009


This image is part of a group.


Image Type
recorded image
Imaging Mode
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
Processing History
unprocessed raw data
color combine
Data Qualifiers
raw, unprocessed data
suitable for spatial measurements

Sample Preparation

living tissue
Relation To Intact Cell
dispersed cells in vitro


Spatial Axis Image Size Pixel Size
X 624px 0.0924μm
Y 302px 0.0924μm
Channel Wavelength
1 488, 561 nm
Time 3,4 sec
*CIL – Cell Image Library accession number. Please use this to reference an image.