Licensing

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Description

Development of the axon and dendritic arbors in cultured hippocampal neurons after 7 days in vitro. MAP2 staining (red) highlights the dendrites, while microtubule staining (green) reveals both the axons and dendrites in the field of view. Neurons at 2, 3, 5 and 7 days in vitro are represented in this image group. Detailed methods: Embryonic rat hippocampal neurons were prepared as previously described (see Kaech and Banker, 2006, Nat Protoc). Cells were prepared for fluorescent staining as previously described (Withers and Banker, 1998, in Culturing Nerve Cells, MIT Press). Briefly, cells were fixed (4% formaldehyde, 4% sucrose in phosphate buffered saline, pH 7.4), permeabilized with 0.25% Triton and immunostained for tubulin (monoclonal DM1A, from Sigma with Alexa 488 conjugated secondary, excitation, 494, emission, 519 [Invitrogen, Molecular Probes]) and MAP2 (polyclonal Ab266, from S. Halpain with d549 conjugated secondary, excitation, 555, emission, 568, Jackson Immunoresearch). Images were acquired with a Leica DMRA microscope with a 20X (Fluotar, NA 0.5) lens, Photometrics CoolSnap ES CCD camera and MetaMorph software. Image generated with the MetaMorph color combine function.

Biological Sources

NCBI Organism Classification
Rattus
(Old World rats)
Cell Type
multipolar neuron
Cellular Component
microtubule cytoskeleton
dendrite
axon

Attribution

Name
Dieter Brandner;Ginger Withers

Grouping

This image is part of a group.

Imaging

Image Type
recorded image
Imaging Mode
fluorescence microscopy
Parameters Imaged
fluorescence emission
Visualization Methods
primary antibody plus labeled secondary antibody
Processing History
unprocessed raw data

Sample Preparation

Methods
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
dispersed cells in vitro

Dimensions

Spatial Axis Image Size Pixel Size
X 1300px 0.339μm
Y 1030px 0.339μm
*CIL – Cell Image Library accession number. Please use this to reference an image.