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

CIL:10105*  Cite 
Description

Human retinal pigmented epithelial (RPE) cells labeled for clathrin-coated pits (green), focal adhesions (red) and nuclei (blue). RPE cells stabily expressing 'clathrin light chain a' tagged with EGFP (EGFP-LCa) were plated directly on glass coverslides. After 72 hrs, cells were fixed for 2 min in a 1:1 mixture of 2% paraformaldehyde/0.5% Triton X-100, followed by a 30 min fixation in 4% paraformaldehyde. Cells were then stained with mouse monoclonal antibodies to paxillin, detected with goat anti-mouse Alexa 568. Nuclei were stained with DAPI. Image was collected on an Olympus IX71 inverted microscope coupled to a Hamamatsu ORCA-ER digital camera using a 60x 1.42 NA Plan Apo N objective (Olympus). The exposure times were 200, 500 and 70 msec for the red, green and blue channel, respectively. A xenon-arc lamp (Lambda LS stand-alone illuminator; Sutter Instrument) was used with adequat filters (Chroma). Bar: 5µm

Biological Sources
NCBI Organism Classification
Homo sapiens
Cell Type
epithelial cell
retinal pigmented epithelial cell
Cellular Component
clathrin coat of coated pit
nucleus
focal adhesion
clathrin light chain a
Attribution
Names
Marcel Mettlen
Sandra L. Schmid
OTHER
The Scripps Research Institute
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL10105
Archival Resource Key (ARK)
ark:/b7295/w9cil10105
Imaging
Image Type
recorded image
Image Mode
fluorescence microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
EGFP
Alexa Fluor 568
4',6-diamidino-2-phenylindole (DAPI)
primary antibody plus labeled secondary antibody
Processing History
3 color combine
Sample Preparation
Methods
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
dispersed cells in vitro
Dimensions
Spatial Axis Image Size Pixel Size
X 1344px 0.027µm
Y 1024px 0.027µm
Channel Wavelength
1 350, 488, 568µm