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 2 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).
To characterize glial morphology and molecular markers in a mouse model of glioblastoma
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
Dinorah Friedmann-Morvinski / Eric Bushong
Microscopy product ID
Nikon RTS 2000 multiphoton microscope
creGFAP mouse injected in the cortex with Ras/s.i.p53 virus and perfused after 2 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-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
nestin labeling of creGFAP brain 2 weeks post inj with Ras/s.i.p53 virus into cortex
Inder Verma, Dinorah Friedmann-Morvinski, Eric Bushong, Mark Ellisman (2007) CCDB:8171, mus musculus. CIL. Dataset. https://doi.org/doi:10.7295/W9CCDB8171