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

CIL:34739*  Cite 

Maximum projection of a multipanel, tiled image through the CA region of the hippocampus of adult rat, immunostained to reveal dendrites of neurons (red), astroglia (green) and all nuclei (blue). This kind of approach can reveal the spatial relationships between the nonneuronal glial cells and neurons.

Technical Details

Tissue was perfused with 4% paraformaldehyde in PBS, postfixed overnight, sectioned at a thickness of 25 µm,incubated in blocking solution (0/25% lambda carageenen, 1%bovine serum albumin, and 0.5% Triton X-100 in 0.01M PBS) and immunostained using antibodies to the dendritically-localized microtubule protein MAP2 (polyclonal Ab266 from S. Halpain), and the astroglial cytoskeletal protein GFAP (glial fibrillary acidic protein, monoclonal clone G-A-5, Sigma) followed by fluorescently-conjugated secondary antibodies (anti-mouse Alexa 488, 1:400; anti-rabbit Alexa 546, both from Molecular Probes), and incubated in 1:1000 Hoescht33258 to stain nuclei. A multiframe data set of 5x3x20(X,Y,Z) was acquired using the Leica SP5 confocal microscope with line-by-line sequential scanning, a 63X HCX PL APOCHROMAT, 1.32NA with 1.7 zoom, at z-intervals of 0.5 µm, and saved as a tif image.

Biological Sources
NCBI Organism Classification
Cell Type
multipolar neuron
glial cell
Cellular Component
intermediate filament cytoskeleton
Biological Context
Biological Process
cell communication by chemical coupling
Christopher Wallace
Ginger Withers
Tony Cooke
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Image Type
recorded image
Image Mode
confocal microscopy
Parameters Imaged
fluorescence microscopy
Source of Contrast
distribution of epitope
differences in adsorption or binding of stain
Visualization Methods
Hoechst 33258
Alexa Fluor 488
Alexa Fluor 546
Processing History
unprocessed raw data
Data Qualifiers
raw, unprocessed data
suitable for spatial measurements
Sample Preparation
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
vibratome-sectioned tissue
Spatial Axis Image Size Pixel Size
X 3251px 0.28µm
Y 1449px 0.28µm