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

CIL:39236*  Cite 
Description

Shown are 0.5 micrometer thick confocal image of midplane sections of synchronized 3T3 fibroblasts pulsed for 2 min with CldU and labeled with FITC (green) followed by a variable chase time, and a second pulse with IdU and subsequently labeled with Cy-5 (red). With simultaneous pulses, the green and red signals overlap (A), but with longer chases, the two sets of replication sites become increasingly separate (B-F).

Technical Details

Cells were fixed with 70% ethanol at -20C, DNA denatured with 4N HCl for 30 min at room temperature, and processed for immunofluorescence using anti-CldU antibody and an appropriate FITC labeled secondary antibody. IuD was detected with a Cy-5 conjugated secondary antibody. Cells were examined in a MRC-1024 confocal microscope mounted on a Nikon Optphot 2 microscope with 60x 1.4 NA objective lens. See: Fig 3 in, Ma, H et al. 1998 Spatial and temporal dynamics of DNA replication sites in mammalian cells. J Cell Biol 143:1415-1425.

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
fibroblast
Cell Line
3T3-Swiss albino
Cellular Component
chromatin
Biological Context
Biological Process
DNA replication involved in S phase
DNA metabolic process
Attribution
Names
Hong Ma
Jagath Samarabandu
Recandu Devdhar
Raj Acharya
Ping-chin Cheng
Chunling Meng
Ronald Berezney
Published
Ma, H et al. 1998 Spatial and temporal dynamics of DNA replication sites in mammalian cells. J Cell Biol 143:1415-1425
Pubmed
9852140
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL39236
Archival Resource Key (ARK)
ark:/b7295/w9cil39236
Grouping This image is part of a group.
Imaging
Image Type
photomultiplier tube (PMT)
Image Mode
confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of epitope
Visualization Methods
Fluorescein (FITC)
TexasRed
Processing History
montage
Data Qualifiers
processed data
Sample Preparation
Methods
ethanol fixed tissue
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 1032px ——
Y 681px ——