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

CIL:11989*  Cite 
Description

TIRF time-lapse imaging of myosin-GFP (left) and mCherry-tubulin (right) in an S2 cell depleted of kinesin-6 (Pav) by RNAi. Myosin-GFP localization to the equator and loss from the poles does not occur after RNAi kinesin-6; microtubules tend to be more homogeneous throughout the cortex and less bundled. Video corresponds to Fig 3B and video 6 from J Cell Bio 186:727-738, 2009. Note that the wild type control is available as another video in the series. 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
myosin regulatory light chain
spindle
Biological Context
Biological Process
mitotic anaphase
kinesin-6 depletion
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/W9CIL11989
Archival Resource Key (ARK)
ark:/b7295/w9cil11989
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
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 452px 0.096µm
Y 226px 0.096µm
Channel Wavelength
1 488, 561nm
Time 4 seconds