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

CIL:41104*  Cite 
Description

Dynamics of membrane ruffling (top) and intracellular vesicle motion (bottom) in a cSrc-transfected COS-7 cell. Images were collected with a Bessel Beam Microscope using two photon excitation. This form of microscopy provides extraordinary 4D resolution, including the individual "slices" shown in the lower right hand panel. This video corresponds to Figure 4c and Supplementary Video 6 in Nat Methods. 2011 May;8(5):417-23. Epub 2011 Mar 4. The video is a deconvolved, maximum projection, volume rendering of slices that were collected in a 48 x 46 x 42 um3 image volume with 116 x 116 x 150 nm3 voxel sizes. The video has a frame rate of 37 msec, and a volume is collected every 10.2 sec.

Technical Details

Scanned Bessel beams can be used to generate thin light sheets that, unlike Gaussian beams, can be decoupled from their longitudinal extent. This video is from a manuscript that describes the use of scanned Bessel beams of higher NA to create light sheets sufficiently thin to achieve isotropic 3D resolution and improve the expenditure of the photon budget to the point at which hundreds of 3D image stacks comprising tens of thousands of frames can be acquired from single living cells at rates of nearly 200 frames/sec.

Biological Sources
NCBI Organism Classification
Chlorocebus aethiops
Cell Line
COS-7
Cellular Component
plasma membrane
ruffle membrane
endocytic vesicle
c-Src
Biological Context
Biological Process
vesicle formation
endocytosis
vesicle organization
Attribution
Names
Thomas A Planchon
Liang Gao
Daniel E Milkie
Michael W Davidson
James A Galbraith
Catherine G Galbraith
Published
Nat Methods. 2011 May;8(5):417-23. Epub 2011 Mar 4.
Pubmed
21378978
Link
article
OTHER
Eric Betzig
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL41104
Archival Resource Key (ARK)
ark:/b7295/w9cil41104
Imaging
Image Type
recorded image
Image Mode
Bessel Sheet Microscpy
Two-Photon Microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
EmeraldFP
Processing History
maximum likelihood deconvolution
maximum intensity projection
volume rendering
Sample Preparation
Methods
living tissue
Relation To Intact Cell
dispersed cells in vitro
Dimensions


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