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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
Cellular Component
plasma membrane
ruffle membrane
endocytic vesicle

Biological Context

Biological Process
vesicle formation
vesicle organization


Thomas A Planchon
Liang Gao
Daniel E Milkie
Michael W Davidson
James A Galbraith
Catherine G Galbraith
Nat Methods. 2011 May;8(5):417-23. Epub 2011 Mar 4.
Eric Betzig


Image Type
recorded image
Imaging Mode
Bessel Sheet Microscpy
Two-Photon Microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
Processing History
maximum likelihood deconvolution
maximum intensity projection
volume rendering

Sample Preparation

living tissue
Relation To Intact Cell
dispersed cells in vitro


Spatial Axis Image Size Pixel Size
X —— ——
Y —— ——
*CIL – Cell Image Library accession number. Please use this to reference an image.