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Reconstruction
Display image description
Large scale brain mosaic showing the distribution of DARRP-32 (green) and VMAT-2 (red) in a hemisection through the anterior neostriatum. To explore the full resolution dataset, click on the "N" in the thumbnail window. This will open up the virtual microscope image explorer.
Full resolution image description
Tiff image of the Z projection of the processed mosaic (~1.7 Gb)
Volume_dimension
20325, 29989, 1
Volume scale
0.24, 0.24, 9

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

CCDB:3592*  Cite 
Project: P1207
Project name
Correlative microscopic characterization of dendritic spines in a transgenic mouse model of hyperdopaminergia: The dopamine transporter knockout mouse
Description
Multiscale characterization of DAT KO transgenic mouse
Funding agency
NIH
Leader(s)
Diana Price
Collaborator(s)
Aki Laakso
Michele Cyr
Maryann Martone
Naoko Yamada
Andrea Thor
Monica Berlanga
Start date
01-01-2003
End date
unspecified
 
Experiment
Experiment ID
3412
Experiment date
09-17-2003
Title
P1207 Exp 3
Purpose
Immunocytochemical localization of VMAT+DARPP-32
Experimenter(s)
Diana Price
Microscopy product
Microscopy product ID
3592
Instrument
BioRad RTS 2000 Multiphoton
Microscopy type
multiphoton
Product type
MOSAIC
Image basename
030304a
Spatial Axis Image Size Pixel Size
X 480px 0.24 um/pixels
Y 480px 0.24 um/pixels
Subject
Species
mouse
Scientific name
mus musculus
Strain
C57BL6/129SvJ
Group by
Genetic manipulation
Treatment
none
Age class
adult
Tissue section
Anatomical location
striatum
Microtome
vibratome
Tissue product storage
Slide box: DAT KO
Thickness
80 µm
Specimen description
Organ
brain
System
central nervous system
Imaging parameters
Type
Light microscopy product
Immersion medium
oil
Mounting medium
gelvatol
Lens
Nikon Plan Apo
Numerical aperture
1.45
Specimen preparation
Protocol used
P1207: Experiment #3DAT KO Mouse9/17/03 Description: Immunolabeling study of VMAT+DARPP-32+Hoescht 33342Animals: Brains sent from Duke University 9/10/03 (wt 1&2, tg 1&2 = 4 total)Protocol1. Perfusion (at Duke U.)Nembutal; 4% paraformaldehyde + 0.1% gluteraldehydeSectioned on Vibratome at NCMIR (80 microns)2. Wash 3x with PBS 1X (on ice) 3x @ 10min1 1st1 2nd1 3r3. Make up blocking bufferPBS w/o NaCl = buffer usedTotal amount needed = 33 x 2 mlsDouble the following:IngredientAmount0.8 PBS6.6 ml 5X PBS + 24.2 ml 2x distilled H203% NDS (24 , 7/4 )0.96 ml 1% fish gel3.3 ml0.3% Triton X-1000.0996 ml1% BSA0.33 g4. Block slices (2 hr) in blocking bufferTime started = 12:40 pm 9/17/03Time ended = 2:50 pm 9/17/035. Make up working buffer" Use blocking buffer to dilute to working bufferIngredient500ml200ml150ml100mlBlocking buffer50 ml20 ml15 ml10 ml0.1% Triton0.5ml0.2 ml0.15 ml0.1 ml1X PBS450 ml180 ml135 ml90 ml6. Wash 1X5 minutes with working buffer: 1 7. Add 1o Abs diluted in working bufferanti-VMAT-2; Host = guinea pig; 1:500 (Oncogene, catalog # 503-01-50)anti-DARPP-32; Host = mouse; 1:500 (BD Transduction Laboratories, catalog #611520)8. Place on shaker in cold room labeled & covered with aluminum foil overnightTime started = 4:30 pm 9/17/03Time ended = 10:30 am 9/18/039. Wash 3x with working buffer 3x @ 10min1 1st1 2nd1 3rdh10. Prepare 2o Abs : all 1:100donkey mouse AF488 (Molecular Probes, Cat #A21202)donkey guinea pig RRX (Jackson Immunoresearch Laboratories, Inc)11. Let sit on shaker covered with foil for 2 hrs at RTTime started = 12:35pm 9/18/03Time ended = 2:50pm 9/18/0312. Wash 3x with 1X PBS 0.8 3x @ 5min1 1st 1 2nd1 3rd13. Prepare nuclear stain (Hoescht 1:1000 for 15-30 minutes)14. Wash 3x with 1X PBS 0.8 3x @ 10min1 1st1 2nd1 3rd15. Mount sections on slides and coverslip using gelvatol16. Dry flat in fridge 24-48 hours and seal with nail polish
Imaging product type
Type
Mosaic
Description
10% overlap between tiles
X position
45 tiles
Y position
65 tiles
Imaging product type
Type
Optical section
Z resolution
3 um