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 5 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 GFAP (red) and counterstained with DAPI (blue). See MP:8063 for a section from the same brain labeled with nestin.
To characterize glial morphology and molecular markers in a mouse model of glioblastoma
Three cre-GFAP mice were injected in the cortex with Ras / s.i. p53 virus. The mice were perfused 5 weeks after injection.
Dinorah Friedmann-Morvinski / Eric Bushong
Microscopy product ID
Nikon RTS 2000 multiphoton microscope
All animals received the same treatment.
creGFAP mice were injected in the cortex with Ras/s.i.p53 virus and perfused after 5 weeks.
central nervous system
Light microscopy product
Nikon 40X oil
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-GFAP antibody (Neuromics Cat#CH22102). 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
GFAP labeling of creGFAP brain 5 weeks post inj in cortex with Ras/s.i.p53 virus
Inder Verma, Dinorah Friedmann-Morvinski, Eric Bushong, Mark Ellisman (2007) CCDB:7857, mus musculus. CIL. Dataset. https://doi.org/doi:10.7295/W9CCDB7857