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

CIL:13757*  Cite 
Description

The Hedgehog (Hh) pathway is activated in follicle stem cells of animals lacking boi (boi[e]; this image), relative to wild-type Drosophila (CIL# 13756), based on activated cubitus interruptus (ci) staining (green). Follicle cells are highlighted with anti-Fas-3 (red) and germ cells are highlighted with anti-vasa (blue). Image is Supplemental Figure S3D in J Cell Biol. 2010. 191: 943-952.

Technical Details

Fly ovaries were dissected and fixed as described previously (O’Reilly et al., 2008). Primary antibodies were 1:2,000 rabbit anti-Vasa (Hay et al., 1990); 1:25 mouse anti-Fas3 (Developmental Studies Hybridoma Bank [DSHB] and anti-activated ci. Secondary antibodies used were FITC, Cy3, and Cy5 conjugated to species-specific secondary antibodies (Jackson ImmunoResearch Laboratories, Inc.). Samples were mounted in Vectashield mounting medium. Images were collected at room temperature (22C) using 40× (1.25 NA) or 63× (1.4 NA) oil immersion lenses (Leica) on an upright microscope (DM 5000; Leica) coupled to a confocal laser scanner (TCS SP5; Leica). LAS AF SP5 software (Leica) was used for data acquisition. Images representing individual channels of single confocal slices from the center of each germarium were exported as TIFF files, and images were converted to figures using Photoshop software (Adobe).

Biological Sources
NCBI Organism Classification
Drosophila melanogaster
Cell Type
germ line cell
follicle cell
germ line stem cell
follicle stem cell
Cell Line
boi[e]
Cellular Component
P granule
plasma membrane
cytoplasm
membrane
Hedgehog signaling complex
Attribution
Names
Tiffiney R. Hartman
Daniel Zinshteyn
Heather K. Schofield
Emmanuelle Nicolas
Ami Okada
Alana M. O'Reilly
Published
J Cell Biol. 2010. 191: 943-952.
Pubmed
21098113
Link
http://jcb-dataviewer.rupress.org/jcb/browse/...
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL13757
Archival Resource Key (ARK)
ark:/b7295/w9cil13757
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
single-spot confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
primary antibody plus labeled secondary antibody
Fluorescein (FITC)
Cy3
Cy5
Processing History
unprocessed raw data
Data Qualifiers
raw, unprocessed data
suitable for spatial measurements
Sample Preparation
Methods
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 512px 0.2327µm
Y 512px 0.2327µm
Z 1px 0.3357µm