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

CIL:10355*  Cite 
Description

Cultured hippocampal neurons after 14 days in vitro, immunostained for MAP2, a microtubule associated protein localized to dendrites (red) but not axons, which are not apparent in the immunofluorescence channel. Both axons, and dendrites, can be seen in the hidden phase micrograph which can be turned on using the edit function in the detailed viewer. Neurons at 10, 14, and 17 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 MAP2 (HM2, from Sigma with d549 conjugated secondary, excitation, 555, emission, 568, Jackson Immunoresearch). Images were acquired with a Leica DMRA microscope with a 40X lens (HCX PL Fluotar, NA 0.75), Photometrics CoolSnap ES CCD camera and MetaMorph software. A multilayer stack of the fluorescent image of MAP2 staining and the phase image was generated using MetaMorph.

Biological Sources
NCBI Organism Classification
Rattus
Cell Type
multipolar neuron
Cellular Component
dendrite
microtubule cytoskeleton
synapse part
Attribution
Names
Dieter Brandner; Ginger Withers
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL10355
Archival Resource Key (ARK)
ark:/b7295/w9cil10355
Grouping This image is part of a group.
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 0.167µm
Y 1030px 0.167µm