8063
Dinorah Friedmann-Morvinski / Eric Bushong
purpose
Batch 14
<![CDATA[http://ccdb.ucsd.edu/sand/main?event=rd&type=512r&mpid=8063]]>
/telescience/home/CCDB_DATA_USER.portal/P1721/Experiment_7596/Subject_7848/Tissue_7850/Microscopy_8063/BT14BR1_NESTIN-1.IMG
1 #
oil
Nikon 40X oil
x
gelvatol
ebushong
1.30
nestin labeling of creGFAP brain 5 weeks post inj with Ras/s.i.p53 virus into cortex
46 tiles
36 tiles
2010-06-15 00:00:00.0
8063
BT14BR1_NESTIN-1.IMG
Nikon RTS 2000 multiphoton microscope
MULTIPHOTON
MOSAIC
.36 um/pixels
512
.36 um/pixels
512
11
Dinorah Friedmann-Morvinski
Eric Bushong
Mark Ellisman
NIH
Inder Verma
Glial markers in mouse model of glioblastoma
2007-06-01 00:00:00.0
P1721
Friedmann-Morvinski D, Bushong EA, Ke E, Soda Y, Marumoto T, Singer O, Ellisman MH, Verma IM, Dedifferentiation of neurons and astrocytes by oncogenes can induce gliomas in mice. Science. 2012 Nov 23;338(6110):1080-4. doi: 10.1126/science.1226929. [PMID:23087000]
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 nestin (red) and counterstained with DAPI (blue). See MP:7857 for a section from the same brain labeled with GFAP.
<![CDATA[http://ccdb.ucsd.edu/sand/main?event=rd&type=512v&mpid=8063]]>
/telescience/home/CCDB_DATA_USER.portal/P1721/Experiment_7596/Subject_7848/Tissue_7850/Microscopy_8063/BT14BR1_NESTIN-1_N.tif
TIFF
22072,16366,1
.36,.36,30
brain
central nervous system
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.
nestin
adult
unspecified
mus musculus
mouse
C57Bl/6
brain
vibratome
70 um
All animals received the same treatment.
creGFAP mice were injected in the cortex with Ras/s.i.p53 virus and perfused after 5 weeks.