The Cell: An Image Library

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CIL:7439^*

Description

Mouse Embryonic Fibroblasts (MEFs) grown on glass coverslips coated with 10 ug/ml Fibronectin. After 24 hrs cells were fixed in 3% paraformaldehyde and stained with mouse antibody to Paxillin, rabbit antibody to cortactin, followed by anti-mouse Alexa 561 goat antibody for Cortactin (cblue), anti-rabbit Alexa 647 for Paxillin (green) and Alexa 488 phalloidin (red) to stain for F-actin. Images were collected on a Nikon Eclipse imaged with a cooled charge coupled device (CCD; Orca II; Hamamatsu Photonics) using a 60x 1.4 NA Plan Apo PH objective lens (Nikon). Individual exposure times were used for each channel: 488: 200ms, 568: 200 ms, 647: 50 ms.

Biological Sources

NCBI Organism Classification: Mus musculus; (house mouse)
Cell Type: fibroblast; primary cell line cell
Cellular Component: focal adhesion; actin cytoskeleton

Biological Context

Biological Process: cell adhesion; actin filament polymerization
Molecular Function: actin binding; cell adhesion molecule binding

Attribution

Names: Ana M. Pasapera; Clare M. Waterman
Pubmed: 20308429
Other: NHLBI/ NIH

Imaging

Image Type: recorded image
Imaging Mode: fluorescence microscopy
Parameters Imaged: fluorescence emission
Source of Contrast: distribution of a specific protein
Visualization Methods: Alexa Fluor 488; Alexa Fluor 568; Alexa Fluor 647; primary antibody plus labeled secondary antibody
Processing History: unprocessed raw data

Sample Preparation

Methods: formaldehyde fixed tissue; detergent permeabilized
Relation To Intact Cell: dispersed cells in vitro

Dimensions

Spatial Axis	Image Size	Pixel Size
X	1392px	0.107nm
Y	1040px	0.107μm
Channel	Wavelength
1	488 nm
2	568 nm
3	647 nm

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