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Description

Disruption of Microtubules with Nocodazol U2OS human osteosarcoma cells were transduced with 'Cellular Lights' Actin-RFP (Actin-red fluorescent protein) and Cellular Lights MAP4-GFP (MAP4-green fluorescent protein) and imaged the next day. Cells were washed in HBSS and incubated with 1 ug/ml of the DNA stain Hoechst 33342 for 10 minutes then washed with HBSS. Nocodazole, a microtubule inhibitor, was applied to the bath at a final concentration of 10 uM. Images were taken every 20 seconds for 30 minutes. In this video, microtubules appear green, actin microfilaments appear red, and the nucleus is stained blue. Note the loss of the microtubular network (green) whilst the actin cytoskeleton (red) remains intact. Cells were imaged on a Deltavision core microscope with a 40X objective, 1.4 NA. Fluorescence was detected using the appropriate filters (DAPI/FITC/TRITC). The time series was deconvolved using standard software. Scale bar = 20 microns.

Biological Sources

NCBI Organism Classification
Homo sapiens
(human)
Cell Type
epithelial cell
permanent cell line cell
Cell Line
U-2 OS
Cellular Component
microtubule cytoskeleton
actin cytoskeleton
nucleus

Biological Context

Biological Process
actin filament depolymerization

Attribution

Names
Nick Dolman
Rob Batchelor
Penny Melquis
Mike O'Grady
George Hanson
Other
Invitrogen, Eugene OR

Grouping

This image is part of a group.

Imaging

Image Type
recorded image
Imaging Mode
fluorescence microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
differences in adsorption or binding of stain
Visualization Methods
mRFP1
Hoechst 33342
EGFP
Processing History
deconvolution

Sample Preparation

Methods
unfixed tissue
Relation To Intact Cell
dispersed cells in vitro

Dimensions

Spatial Axis Image Size Pixel Size
X 538px 0.20μm
Y 528px 0.20μm
Channel Wavelength
1 DAPI/FITC/TRITC nm
Time 1800 sec 20s/frame
*CIL – Cell Image Library accession number. Please use this to reference an image.