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Title: Cell Image Library Dataset CIL:10273      
dateReleased:
12-08-2010
description:
Early stages of dendritic development and synapse formation in cultured hippocampal neurons. This multilayer image shows neurons fixed at 7 days in vitro and immunostained for the dendritically localized protein MAP2 (green) and the presynaptic vesicle protein Synapsin I (red). Unstained axons in the field are evident in the phase channel, which is hidden but can be turned on in the viewer. Neurons at 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, warmed to 37°C prior to fixation, 15 minutes), permeabilized with 0.25% Triton (7 minutes) and immunostained for MAP2 (monoclonal HM2, Sigma, with Alexa 488 conjugated secondary, excitation, 494, emission, 519 [Invitrogen, Molecular Probes]) and synapsin I (from P. DeCamilli, with DyLight549 conjugated secondary, excitation, 555, emission, 568, [Jackson Immunoresearch]). Images were acquired with a Leica DMRA microscope with a mercury arc lamp, a 40X lens (HCX PL Fluotar, NA 0.75), Leica GFP filter set (excitation, BP 470/40; dichromatic mirror, 500; suppression filter, BP 525/50); Leica N3 filter set (excitation, BP546/12; dichromatic mirror, 565, suppression filter, BP 600/40), Photometrics CoolSnap ES CCD camera and MetaMorph software.
privacy:
not applicable
aggregation:
instance of dataset
ID:
10273
refinement:
curated
availability:
available
types:
image
dimensions:
fluorescence emission
keywords:
primary antibody plus labeled secondary antibody
name:
detergent permeabilized, formaldehyde fixed tissue
name:
multipolar neuron
name:
cellular developmental process
name:
dendrite development
name:
establishment and/or maintenance of cell polarity
name:
establishment or maintenance of cell polarity
name:
protein localisation to synapse
name:
protein localization to synapse
name:
synapse assembly
name:
synapse biogenesis
name:
dendrite
name:
microtubule cytoskeleton
name:
synapse part
name:
fluorescence microscopy, phase contrast microscopy
accessURL: www.cellimagelibrary.org/images/download_jpeg/10273.jpg
storedIn:
Cell Image Library
qualifier:
not compressed
license:
attribution_cc_by
format:
JPGE
accesstypes:
download
accessType:
download
authentication:
none
authorization:
none
accessURL: http://grackle.crbs.ucsd.edu:8080/OmeroWebService/images/10273.tif
storedIn:
Cell Image Library
qualifier:
not compressed
license:
attribution_cc_by
format:
OME-TIF
accesstypes:
download
accessType:
download
authentication:
none
authorization:
none
accessURL: http://grackle.crbs.ucsd.edu:8080/OmeroWebService/images/10273.raw
storedIn:
Cell Image Library
qualifier:
compressed
license:
attribution_cc_by
format:
CSV
accesstypes:
download
accessType:
download
authentication:
none
authorization:
none
abbreviation:
ASCB
homePage: http://ascb.org/
ID:
SCR:010600
name:
American Society for Cell Biology
abbreviation:
CIL
homePage: http://www.cellimagelibrary.org/
ID:
SCR:003510
name:
Cell Image Library

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