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

CIL:13677*  Cite 
Description

The JNK phosphatase puckered, whose expression can be reported via a lacZ enhancer trap, is expressed in very low levels in uninjured neurons; however, nerve crush induces a dramatic increase in puc-lacZ expression in injured motoneurons. This image stack shows puc-lacZ (green) and neuronal nuclei (red) in a third instar larva injred at site 3 (as defined in Fig 1C). Wandering third instar larvae were dissected in PBS and fixed in either 4% paraformaldehyde in PBS or Bouin’s fixative for 15–30 min. Primary antibodies used were: rat anti-elaV (7E8A10, Developmental Studies Hybridoma Bank) and mouse anti-lacZ (40-1a, DSHB). Secondary antibodies were Alexa 488 anti-mouse and Cy3 anti-rat. Confocal images were collected at room temperature on a spinning-disk confocal system (PerkinElmer) consisting of a scanner (Nipkow CSU10; Yokogawa) and an electron microscopy charge-coupled device camera (C9100-50; Hamamatsu Photonics) mounted on an inverted microscope (Axio Observer; Carl Zeiss, Inc.) with 25× 0.8 NA multi and 40× 1.3 NA, 63× 1.5 NA, and 100× 1.46 NA oil objectives. Similar settings were used to collect all compared genotypes. All imaging and quantification were conducted with Volocity software (PerkinElmer). This image stack corresponds to Fig 1E3 in J Cell Biol. 2010. 191: 211-223. Images in Fig 1 include CIL#s 13676, 13677, 13678, 13679.

Biological Sources
NCBI Organism Classification
Drosophila melanogaster
Cell Type
motor neuron
Cellular Component
nucleus
Attribution
Names
Xin Xiong
Xin Wang
Ronny Ewanek
Pavan Bhat
Aaron DiAntonio
Catherine A. Collins
Published
J Cell Biol. 2010. 191: 211-223.
Pubmed
20921142
Link
http://jcb-dataviewer.rupress.org/jcb/browse/...
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL13677
Archival Resource Key (ARK)
ark:/b7295/w9cil13677
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
spinning disk confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
primary antibody plus labeled secondary antibody
beta-galactosidase
Alexa Fluor 488
Cy3
Data Qualifiers
raw, unprocessed data
suitable for spatial measurements
Sample Preparation
Methods
formaldehyde fixed tissue
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 962px 0.5869µm
Y 1000px 0.5869µm
Z 8px 0.8µm