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

CIL:34544*  Cite 
Description

Immunohistochemistry of neuron-specific β-III-tubulin (red) shows the delicate dendrites of Purkinje cells in the cerebellum. Cell nuclei are revealed with DAPI staining (blue). Scale bar, 100 µm.

Technical Details

The adult mouse were perfused transcardially with 4% PFA following anesthetization with avertin. The brain was dissected out and fixed by immersion in 4% paraformaldehyde (PFA) overnight at 4 °C, embedded in paraffin and cut into 5-µm-thick sagittal sections, which were deparaffinized using a standard histology protocol immediately before immunohistochemical staining. In the staining procedure, tissue sections were permeabilized with 0.5% Triton X-100 (Sigma) for 10 minutes, then blocked with the buffer containing 10% normal goat serum (Sigma), 1% (w/v) bovine serum albumin (BSA) and 0.2% (v/v) Triton X-100 for 2 hours at RT, followed by incubation with anti-βIII-tubulin monoclonal antibody (Covance) at1:50 dilution in the dilution buffer (2% normal goat serum, 1% BSA, 0.1% Triton X-100) overnight at 4 °C. Samples were subsequently incubated with FITC- and/or Cy3-conjugated species-specific secondary antibody/antibodies in the dilution buffer (1:200 dilution) for 1 hour at RT. VECTASHIELD Mounting Medium with DAPI (Vector Laboratories) was used to mount the fluorescently labeled samples and to stain cell nuclei. Images were digitally acquired using a fluorescence microscope (Nikon Eclipse 660) equipped with Spot cooled CCD camera (Diagnostic Instruments).

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
CNS neuron (sensu Vertebrata)
Purkinje cell
Cellular Component
dendrite
nucleus
Attribution
Names
Yinghua Ma
Timothy Vartanian
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL34544
Archival Resource Key (ARK)
ark:/b7295/w9cil34544
Imaging
Image Type
recorded image
Image Mode
fluorescence microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of epitope
Visualization Methods
Cy3
4',6-diamidino-2-phenylindole (DAPI)
Data Qualifiers
processed data
Sample Preparation
Methods
formaldehyde fixed tissue
tissue in paraffin embedment
Relation To Intact Cell
microtome-sectioned tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 4667px 7.77µm
Y 3500px 7.77µm