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

CIL:40003*  Cite 
Description

Large scale brain mosaic showing the distribution of the dopamine receptor-associated phosphoprotein DARRP-32 (green) and the vesicular monamine transporter VMAT-2 (red) in a hemisection through the anterior neostriatum of a dopamine transporter knock out mouse. This image was generated by tiling 45 by 65 (X, Y axes) frames of maximum projection images in the Z-axis. This image has been downsampled from the raw data image which can be accessed using the link provided to the Cell Centered Database.

Technical Details

Transgenic mice (strain DATKO) from Duke University were perfused with aldehyde fixatives (4% paraformaldehyde + 0.1% gluteraldehyde), sectioned on Vibratome (thickness, 80 µm), and rinsed in phosphate buffered saline (PBS,3 x 10min) and incubated in blocking buffer (PBS, 3% normal donkey serum, 1% fish skin gelatin, 0.3% Triton X-100, 1% bovine serum albumin). Tissue sections incubated on shaker overnight, 4° C in primary antibodies diluted in blocking buffer as follows: anti-VMAT-2; Host = guinea pig; 1:500 (Oncogene, catalog # 503-01-50); anti-DARPP-32; Host = mouse; 1:500 (BD Transduction Laboratories, catalog #611520). Following 3 x 10' rinses, tissue was incubated for 2hr, RT, on a shaker in secondary antibodies diluted at 1:100 in working buffer (donkey-anti-mouse AF488 [Molecular Probes, Cat #A21202] and donkey-anti-guinea pig RRX [Jackson Immunoresearch Laboratories, Inc]. Following rinses in PBS, sections were mounted on slides with gelvatol. Individual images were gathered using a BioRad RTS 2000 Multiphoton with a Nikon PlanApo objective (N.A. 1.45).

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
CNS neuron (sensu Vertebrata)
Attribution
Names
Diana Price
Aki Laakso
Maryann Martone
Naoko Yamada
Andrea Thor
Monica Berlanga
Michele Cyr
Link
CCDB 3593
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL40003
Archival Resource Key (ARK)
ark:/b7295/w9cil40003
Imaging
Image Type
recorded image
Image Mode
multi-photon microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of epitope
Visualization Methods
primary antibody plus labeled secondary antibody
Alexa Fluor 488
X-Rhodamine
Data Qualifiers
processed data
Sample Preparation
Methods
formaldehyde fixed tissue
glutaraldehyde fixed tissue
Relation To Intact Cell
vibratome-sectioned tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 512px ——
Y 512px ——