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CIL:10392*  Cite 

Cultured hippocampal neurons after 23 days in vitro, immunostained for MAP2, a microtubule associated protein localized to dendrites (red), and PSD95, a scaffolding protein concentrated in the postsynaptic density of excitatory synapses, particularly in dendritic spines (green). 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 (from S. Halpain) with d549 conjugated secondary, excitation, 555, emission, 568, Jackson Immunoresearch) and PSD95 (from NeuroMab, with Alexa488 conjugated secondary, excitation, 496, emission, 519, Invitrogen). 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. The merged image was generated by creating a MetaMorph stack file.

Biological Sources
NCBI Organism Classification
Cell Type
multipolar neuron
Cellular Component
microtubule cytoskeleton
synapse part
Dieter Brandner (Whitman College)
Ginger Withers (Whitman College)
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Image Type
recorded image
Image Mode
fluorescence microscopy
phase contrast microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of epitope
elastic scattering of photons
Visualization Methods
primary antibody plus labeled secondary antibody
Processing History
unprocessed raw data
Data Qualifiers
processed data
suitable for spatial measurements
Sample Preparation
formaldehyde fixed tissue
detergent permeabilized
Relation To Intact Cell
dispersed cells in vitro
Spatial Axis Image Size Pixel Size
X 1300px 0.167µm
Y 1030px 0.167µm