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

CIL:40121*  Cite 

Wide field brain mosaic of a coronal section of brain from a transgenic mouse engineered to overexpress human alpha synuclein immunolabeled for alpha synuclein (Green) and counterstained with Hoescht 33342 (red) to reveal cell nuclei. The Hoescht staining introduced some non-specific background (red haze). 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

Transgenic adult female mouse (624 days, 32 g) 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. 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 Gelvatol. Sections were imaged using a BioRad RTS 2000MP Multiphoton, with a 60X, NA 1.4 objective. This tiled image was generated from 57 x 46 (x, y) maximum projection images.

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
CNS neuron (sensu Vertebrata)
D.L. Price
M.H. Ellisman
M. Martone
G.A. Johnson
E. Masliah
CCDB 3993
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
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
Hoechst 33342
Data Qualifiers
processed data
Sample Preparation
formaldehyde fixed tissue
glutaraldehyde fixed tissue
Relation To Intact Cell
vibratome-sectioned tissue
Spatial Axis Image Size Pixel Size
X 512px ——
Y 512px ——