Planchon, Thomas A.; Gao, Liang; Milkie, Daniel E; Davidson, Michael W.; Galbraith, James A.; Galbraith, Catherine G. (2021). CIL:41104, Chlorocebus aethiops. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J09C6W76

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.

• 2021

• Davidson, Michael W.
• Galbraith, Catherine G.
• Galbraith, James A.
• Gao, Liang
• Milkie, Daniel E
• Planchon, Thomas A.

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.

Preparation: living tissue
Relation to intact cell: dispersed cells in vitro
Item type: recorded image
Imaging mode: Bessel Sheet Microscpy; Two-Photon Microscopy
Parameter imaged: fluorescence emission
Source of contrast: distribution of a specific protein
Visualization methods: EmeraldFP
Processing history: maximum likelihood deconvolution; maximum intensity projection; volume rendering

• Chlorocebus aethiops

• C-Src
• Endocytic vesicle
• Plasma membrane
• Ruffle membrane

• COS-7
• Endocytosis
• Vesicle formation
• Vesicle organization

• data
• image
• video

• No linguistic content; Not applicable

Samplenumber: 41104

Doi: https://doi.org/10.6075/J09C6W76

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL41104

Reference

• Korobova F, Svitkina T. Molecular architecture of synaptic actin cytoskeleton in hippocampal neurons reveals a mechanism of dendritic spine morphogenesis. Mol Biol Cell. 2010 Jan 1;21(1):165-76. https://doi.org/10.1091/mbc.e09-07-0596
• Nat Methods. 2011 May;8(5):417-23. Epub 2011 Mar 4. https://www.ncbi.nlm.nih.gov/pubmed/?term=21378978
• Planchon TA, Gao L, Milkie DE, Davidson MW, Galbraith JA, Galbraith CG, Betzig E. Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination. Nat Methods. 2011 May;8(5):417-23. https://doi.org/10.1038/nmeth.1586

Other resource

• George E. Palade EM Slide Collection: http://cushing.med.yale.edu/gsdl/cgi-bin/library?p=about&c=palade

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2022-08-12