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

CIL:13477*  Cite 
Description

Ema localizes to endosomes. Third instar larval garland cells expressing Ema-GFP fusion protein and stained for the late endosomal and lysosomal protein Spinster (red) and plasma membrane and nuclei (blue). Ema co-localizes with some late endosomes. Wandering third instar larvae were dissected in ice-cold PBS, equilibrated in HL-3 buffer at room temperature, incubated in 0.7% HRP for 5 min at room temperature, rinsed thoroughly and fixed with Bouin’s solution for 5 min. Blocking and antibody incubation were performed in PBS containing 0.1% Triton X-100. Anti-Spinster antibody (Sweeney and Davis, 2002) used at 1:250. NeuroTrace 640/660 deep red fluorescent Nissl (Invitrogen) used at 1:500 was used to stain nuclei. Cy5-conjugated anti-HRP (Jackson Immunoresearch Laboratories) at 1:1000 was used to stain plasma membrane. Cy3-conjugated secondary antibody (Jackson Immunoresearch Laboratories, Inc.) at 1:1,000. Confocal images were acquired with a confocal microscope (model C1; Nikon) and accompanying EZ-C1 software using argon (excitation at 488 nm) and HeNe (excitation at 543 and 633 nm) lasers and a 60x Plan-Apochromat NA 1.4 objective (Nikon) at room temperature. Samples for each experiment were processed using the same confocal gain setting. Image corresponds to Fig 3B in Kim et al. J Cell Biol. 188: 717-734. 2010.

Biological Sources
NCBI Organism Classification
Drosophila melanogaster
Cell Type
garland cell
Cellular Component
endosome
late endosome
plasma membrane
nucleus
Biological Context
Biological Process
endocytosis
Attribution
Names
Sungsu Kim
Yogesh P. Wairkar
Richard W. Daniels
Aaron DiAntonio
Published
J Cell Biol. 188: 717-734. 2010
Pubmed
PMID: 20194640
Link
http://jcb-dataviewer.rupress.org/jcb/browse/...
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL13477
Archival Resource Key (ARK)
ark:/b7295/w9cil13477
Grouping This image is part of a group.
Imaging
Image Type
recorded image
photomultiplier tube (PMT)
Image Mode
fluorescence microscopy
confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
distribution of a specific nucleic acid sequence
distribution of epitope
Visualization Methods
EGFP
Cy5
Deep-red fluorescent Nissl stain
Cy3
Data Qualifiers
raw, unprocessed data
Sample Preparation
Methods
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 1024px ——
Y 288px ——