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

CIL:40197*  Cite 
Description

Mosaic image of mouse cerebellum reconstructed through automated acquisition and stitching of over 1700 individual tiles in the X, Y and Z planes, triple labeled for astrocytes (GFAP; Red), gap junctions (Connexin 43; Green) and nuclei (DAPI; Blue), and imaged using multiphoton microscopy. The contrast and brightness were adjusted on the thumbnail image for display purposes. This image has been downsampled from the raw data image which can be accessed using the link provided to the Cell Centered Database.

Technical Details

Mouse brain was perfused with 4% paraformaldehyde, sectioned through the cerebellum in the sagittal plane on a vibratome (50 µm). Tissue sections were immunostained using Rabbit Cx43 antibody (Sigma) 1:600, and Chicken GFAP antibody (Neuromics) 1:500, 72hour incubation at 4C. Secondary antibodies were Donkey anti rabbit-Al488, and Donkey anti-chicken-RRX, 2.5hour incubation at room temperature, followed by staining with DAPI. Sections were mounted in gelvatol. Individual images were gathered using a Nikon PlanApo 60X NA 1.45 objective on a BioRad RTS2000 multiphoton microscope. The mosaic image was generated by tiling 37 (X) x 48 (Y) maximum projection images.

Biological Sources
NCBI Organism Classification
Mus musculus
Cell Type
CNS neuron (sensu Vertebrata)
astrocyte of the cerebellum
Cellular Component
intermediate filament
gap junction
nucleus
Attribution
Names
Angela Cone
Gina Sosinsky
Maryann Martone
Link
CCDB 6624
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL40197
Archival Resource Key (ARK)
ark:/b7295/w9cil40197
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
multi-photon microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of epitope
Visualization Methods
primary antibody plus labeled secondary antibody
Alexa Fluor 488
X-Rhodamine
4',6-diamidino-2-phenylindole (DAPI)
Data Qualifiers
processed data
Sample Preparation
Methods
formaldehyde fixed tissue
Relation To Intact Cell
vibratome-sectioned tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 512px ——
Y 621px ——


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