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

CIL:40004*  Cite 
Description

Large scale brain mosaic at the level of the anterior hippocampus showing the localization of alpha synuclein (green) in a normal mouse. Section was counterstained with a nuclear stain (blue) to reveal the locations of cell somata. This mosaic image has been downsampled from the raw data image, which can be accessed using the link provided to the Cell Centered Database. For more information regarding this project, see: Price DL, Chow SK, MacLean NAB, Hakozaki H, Peltier S, Martone ME, Ellisman MH (2006) High-Resolution Large-Scale Mosaic Imaging using Multiphoton Microscopy to Characterize Transgenic Mouse Models of Human Neurological Disorders. Neuroinformatics. 2006;4(1):65-80.

Technical Details

Wild type adult male mouse was anesthetized with nembutal, and perfused with 4% paraformaldehyde + 0.1% glutaraldehyde, followed by 1 hr. postfixation in 4% paraformaldehyde. Tissue was sectioned on a vibratome at a thickness of 80µm. Following rinses in phosphate buffer (PBS), sections were blocked (PBS with 3% NGS;1% fish gel, 0.1% Triton X-1000; 1% BSA) for 1 hour, followed by rinses in working buffer (1:10 blocking buffer: PBS). Primary antibody was diluted in working buffer (anti-alpha-SYN; Host = Rabbit used at 1:500), and tissue was covered with aluminum foil and incubated on a shaker in cold room overnight. Tissue was washed with working buffer and incubated in secondary antibody (goat anti-rabbit AlexaFluor 568, 1:50) covered with foil, on cold room shaker, 48 hrs. After rinses, 1:100 Hoescht 33342 was used to stain nuclei (30 min). After rinsing in PBS, sections were mounted on slides and coverslipped using Vectashield. Sections were imaged using a BioRad RTS 2000MP Multiphoton, with a Nikon Plan Fluor 60X, NA 1.4 objective.

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
CNS neuron (sensu Vertebrata)
Attribution
Names
D. Price
M.H. Ellisman
M. Martone
G.A. Johnson
E. Masliah
Link
CCDB 36
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL40004
Archival Resource Key (ARK)
ark:/b7295/w9cil40004
Grouping This image is part of a group.
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 568
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 480px ——