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

CIL:34930^* Cite

Description

Localization of cross-linking proteins in fibroblast cytoskeleton. Immuno-EM of the cell interior at cell edge (CIL 34926) of cytochalasin D-treated Xenopus fibroblasts stained with α-actinin primary antibody and 18-nm gold-conjugated secondary antibody. Gold particles (yellow) reveal α-actinin at filament crossovers in the cell interior. A fluorescence image of fibroblast lamellipodia double stained with TRITC-phalloidin (red) and either α-actinin (green) is available at CIL 34924. The image corresponds to Figure 5f from J Cell Biol. 1999 May 31;145(5):1009-26.

Technical Details

Procedures for detergent extraction, immunostaining, S1 decoration, light, and EM were described previously (Svitkina et al., 1995, 1996, 1997;Verkhovsky et al., 1995; Svitkina and Borisy, 1998).

Biological Sources

NCBI Organism Classification: Xenopus laevis
Cell Type: fibroblast
Cellular Component: actin cytoskeleton; alpha actinin; cell interior

Biological Context

Biological Process: branching of actin filaments; cytochalasin D treatment; linking of actin filaments
Molecular Function: actin filament-based process

Attribution

Names: Tatyana M. Svitkina; Gary G. Borisy
Published: J Cell Biol. 1999 May 31;145(5):1009-26.
Pubmed: 10352018

Citation

Digital Object Identifier (DOI): doi:10.7295/W9CIL34930
Archival Resource Key (ARK): ark:/b7295/w9cil34930

Grouping This image is part of a group.

Imaging

Image Type: recorded image
Image Mode: transmission electron microscopy (TEM)
Parameters Imaged: optical path length gradient
Source of Contrast: differences in deposition of metal shadow
Visualization Methods: shadowing and plating; platinum replica; primary antibody plus labeled secondary antibody; immuno-gold
Processing History: pseudocolored

Sample Preparation

Methods: permeabilized tissue; glutaraldehyde fixed tissue; phalloidin; methanol fixed tissue
Relation To Intact Cell: dispersed cells in vitro

Dimensions

Spatial Axis	Image Size	Pixel Size
X	780px	2.75nm
Y	661px	2.75nm