Alternate header for print version


Reconstruction
Display image description
Processed mosaic of a sagittal section through the brain of mouse model of glioma. To produce transformed cells, creGFAP mice were injected in the cortex with Ras/s.i.p53 virus and perfused after 8 weeks. In this system, GFP is introduced by the virus and expression is induced by an IRES site in the mRNA. Thus, the GFP gene is expressed whenever the virus infects a cell expressing GFAP (green). The section was immunolabeled for nestin (red) and counterstained with DAPI (blue). See MP:8011 for a section from the same brain labeled with GFAP.
File format
MRC
Volume_dimension
32149, 22250, 1
Volume scale
0.36, 0.36, 33

License
Attribution Only: This image is licensed under a Creative Commons Attribution License. View License Deed | View Legal Code

CCDB:8166*  Cite 
Project: P1721
Project name
Glial markers in mouse model of glioblastoma
Description
To characterize glial morphology and molecular markers in a mouse model of glioblastoma
Funding agency
NIH
Leader(s)
Inder Verma
Collaborator(s)
Dinorah Friedmann-Morvinski
Eric Bushong
Mark Ellisman
Start date
06-01-2007
End date
unspecified
 
Experiment
Experiment ID
7804
Title
Batch 19
Purpose
Batch 19 consists of 8 brains of different experimental treatments. Brains 1 & 2: GFAP-cre mice injected in cortex with Ras / si p53 virus and perfused 8 weeks after injection Brains 3,4,& 5: nestin-cre mice injected in the hippocampus with Ras / si p53 virus and perfused 2 weeks after injection Brains 6,7,& 8: GFAP-cre mice injected in cortex with Ras/ si p53 virus and perfused 2 weeks after injection
Experimenter(s)
Dinorah Friedmann-Morvinski / Eric Bushong
Microscopy product
Microscopy product ID
8166
Instrument
Nikon RTS 2000 multiphoton microscope
Microscopy type
MULTIPHOTON
Product type
MOSAIC
Image basename
BT19BR2_NESTIN-1.IMG
Spatial Axis Image Size Pixel Size
X 512px 0.36 um/pixels
Y 512px 0.36 um/pixels
Y 11px
Subject
Species
mouse
Scientific name
mus musculus
Strain
C57Bl/6
Treatment
Brain #2 was from a creGFAP mouse injected in the cortex with Ras/s.i.p53 virus and perfused after 8 weeks.
Age class
adult
Tissue section
Anatomical location
brain
Microtome
vibratome
Thickness
70 µm
Specimen description
Organ
brain
System
central nervous system
Imaging parameters
Type
Light microscopy product
Immersion medium
oil
Mounting medium
gelvatol
Lens
Nikon 40X oil
Lens magnification
x
Numerical aperture
1.30
Notes
ebushong
Specimen preparation
Protocol used
A slice was taken from cryoprotectant and washed for 3x 10 minutes in PBS.The slice was placed in blocking buffer at room temp for 1 hour.Blocking buffer: PBS, 3% NDS, 1% BSA, 1% CWFG, 0.3% Triton X-100The slice was briefly washed in working buffer.Working buffer: 1 in 10 dilution of blocking buffer in PBS, plus 0.1% tritonThe slice was placed in working buffer containing 1:500 dilution of chicken anti-nestin antibody (Neuromics Cat#CH23001, Lot#400588). The slice was left in cold room overnight.The slice was washed 3x 20 mintues in working buffer at room temp.The slice was placed in working buffer containing 1:200 dilution of donkey anti-chicken RRX antibody. The slice was left in cold room overnight.The slice was washed 1X 15 minutes in PBS.The slice was placed in DAPI/PBS solution for 15 minutes.The slice was washed 2x 15 minutes in PBS.The slice was coverslipped in Gelvatol and stored in fridge until imaged.
Imaging product type
Type
Mosaic
Description
Nestin labeling of Cre-GFAP mouse perfused 8 weeks after injection in cortex with Ras/s.i. p53
X position
67 tiles
Y position
49 tiles