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

CIL:7429*  Cite 
Description

Image shows (actin polymerization at) free barbed ends and F-actin in the lamellipodium of a migrating primary murine embryonic fibroblast. Cell has been mildly permeabilized using saponin during a 2min incubation in cytoskeleton buffer containing X-rhodamine labeled G-actin. Intracellular X-rhodamine actin polymerized into filaments at the very leading edge of the lamellipodium, resulting in the narrow red band along the tip of the lamelipodium. Lamellipodial F-actin was labeled with Alexa488-phalloidin using glutaraldehyde fixation subsequently to the X-rhodamine actin incubation. The micrograph visualizes that the actin monomer incorporation into the lamellipodial actin filament structure is restricted to the front of the lamellipodium. This enables a directed extension of the branched actin network towards the plasmamembrane, which pushes the membrane forward thereby creating cell protrusion. Wide field epifluorescence image taken on using a 100X 1.49 NA lens, Semrock DA/FI/TR/Cy5-4x4M-B Sedat filter set, and an Orca II with 500 ms exposure.

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
fibroblast
primary cell line cell
Cellular Component
actin cytoskeleton
Biological Context
Biological Process
actin filament polymerization
Molecular Function
actin binding
Attribution
Names
Ingo Thieverssen
Clare M. Waterman
OTHER
Laboratory of Cell and Tissue Morphodynamics
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL7429
Archival Resource Key (ARK)
ark:/b7295/w9cil7429
Imaging
Image Type
recorded image
Image Mode
widefield illumination
fluorescence microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
Alexa Fluor 488
X-Rhodamine
phalloidin
Processing History
unprocessed raw data
Sample Preparation
Methods
glutaraldehyde fixed tissue
saponin permeabilized
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 500px 0.064µm
Y 500px 0.064µm
Channel Wavelength
1 488nm
2 561nm