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Description

Mitochondria in HeLa cells are identified by mito-YFP (green). When fixed cells are permeabilized with 0.005% digitonin, the antibody to cyctochrome c (red), a mitochondrial matrix marker, is unable to reach and bind cytochrome c, but the antibody to Tom20 (white), a mitochondrial outer membrane marker, is now able to bind Tom20. Mito-YFP transfected HeLa cells were treated with DMSO for 3 hrs, fixed with 4% paraformaldehyde and permeabilized with 0.005% digitonin/PBS. Primary antibodies used were: anti-Cyt c mAb (BD) and anti-Tom20 mAb (BD); secondary antibodies used were: Alexa Fluor 594 and 647. Imaging was performed on an LSM510 Meta (Carl Zeiss, Inc) with a 63x 1.4 NA oil differential interference contrast Plan Apo objective. Image contrast and brightness were adjusted in the LSM image browser (Zeiss). This image corresponds to Fig 4b, 2nd row from top and is part of a differential permeabilization assay with and without the mitochondrial depolarizing agent CCCP (carbonyl cyanide m-chlorophenyl hydrazone) that is further described in Fig 4b of J Cell Biol, 191: 933-942, 2010. Images in Fig 4 include CIL#s 13733, 13734, 13729, 13730, 13731, 13732, 13717, 13718, 13719, 13720, 13721, 13722, 13723, 13724.

Biological Sources

NCBI Organism Classification
Homo sapiens
(human)
Cell Type
endothelial cell
Cell Line
HeLa
cervical carcinoma
Cellular Component
mitochondrion
mitochondrial outer membrane translocase complex
mitochondrial intermembrane space

Attribution

Names
Seok Min Jin
Michael Lazarou
Chunxin Wang
Lesley A. Kane
Derek P. Narendra
Richard J. Youle
Published
J Cell Biol, 191: 933-942, 2010.
Link
http://jcb-dataviewer.rupress.org/jcb/...
Pubmed
21115803

Grouping

This image is part of a group.

Imaging

Image Type
recorded image
Imaging Mode
single-spot confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
EYFP
primary antibody plus labeled secondary antibody
Processing History
brightness and contrast adjusted
Data Qualifiers
raw, unprocessed data
suitable for spatial measurements

Sample Preparation

Methods
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
dispersed cells in vitro

Dimensions

Spatial Axis Image Size Pixel Size
X 512px .1860μm
Y 512px .1860μm
*CIL – Cell Image Library accession number. Please use this to reference an image.