3993
2003-02-18 00:00:00.0
Diana Price
purpose
Alpha-synuclein immunolabeling for large-scale mapping study
oil
X
1.4
10% overlap of tile
57 tiles
46 tiles
3 3
3-02-18 00:00:00.0
3993
051903A
BioRad RTS 2000MP Multiphoton
multiphoton
MOSAIC
.237 um/pixels
512
.237 um/pixels
480
M.H. Ellisman; M. Martone; G.A. Johnson; E. Masliah
The Branfman Family Foundation
Diana Price
2003-06-30 00:00:00.0
Correlated Imaging Approaches and Multiscale Databases for Research in Parkinson's Disease
2002-09-01 00:00:00.0
P1187
Price DL;Martone ME; Masliah MH; Ellisman MH (2003) High-resolution Large-Scale 3-D Mapping Studies of Alpha-Synuclein Immunoreactivity in Transgenic Mice Overexpressing Human Alpha-Synuclein. Society for Neuroscience Abstract.
Price DL, Chow SK, MacLean NAB, Hakozaki H, Peltier S, Martone ME, Ellisman MH (2006) High-Resolution Large-Scale Mosaic Imaging using Multiphoton Microscopy to Characterize Transgenic Mouse Models of Human Neurological Disorders. Neuroinformatics. 2006;4(1):65-80.
Wide field brain mosaic of a coronal section of brain from a transgenic mouse engineered to overexpress human alpha synuclein immunolabeled for alpha synuclein (Green) and counterstained with Hoescht 33342 (red) to reveal cell nuclei. The Hoescht staining introduced some non-specific background (red haze).
<![CDATA[http://ccdb.ucsd.edu/sand/main?event=rd&type=512v&mpid=3993]]>
Processed mosaic in .tif format (051903_vol.tif). Processed mosaic has all the tiles assembled and is a projection of the 3 optical sections acquired for each tile. The reconstruction has been rotated from the original 90 degrees CCW and flipped on the horizontal axis to conform with the orientation of a standard brain atlas and other images in this set. The original orientation has been uploaded as well (051903montage.tif) and is available for download upon request to the CCDB.
/telescience/home/CCDB_DATA_USER.portal/P1187/Experiment_17/Subject_17/Tissue_320/Microscopy_3993/051903_vol.tif
20116,26664,1
.24,.24,9
0,0,0
Coronal slice 41
brain
central nervous
Description: alpha-synuclein immunolabeling for large-scale mapping study (non-tg)Animals: 548 (f) transgenic from D-line (see subject log for details)Protocol1. PerfusionNembutal; 4% paraformaldehyde + 0.1% gluteraldehyde1 hr. postfix in 4% para2. Sectioned on VibratomeThickness = 80 microns3 blocks at 2 mm each from anterior (A, B, C) + cerebellumLeft hemisphere marked3. Wash 6x with PBS 1X (on ice)4. Make up blocking bufferPBS w/o NaCl = buffer usedTotal amount needed = 33 mls x 2Ingredient Amount0.8 PBS 6.6 ml 5X PBS + 24.2 ml 2x distilled H203% NGS (24 , 7 ) 0.96 ml1% fish gel 0.33 ml0.1% Triton X-100 0.0332 ml1% BSA 0.33 g5. Block slices (1 hr) in blocking bufferIngredient Amount0.8 PBS 6.6 ml 5X PBS + 24.2 ml 2x distilled H203% NGS (24 , 7 ) 0.96 ml1% fish gel 0.33 ml0.1% Triton X-100 0.0332 ml1% BSA 0.33 g6. Make up working bufferUse blocking buffer to dilute to working bufferIngredient AmountBlocking buffer 20 ml0.1% Triton 0.2 ml1X PBS 180 ml7. Wash 1X5 minutes with working buffer 8. Dilute primary Abs in working bufferanti-alpha-SYN; Host = Rabbit 1:5009. Place on shaker in cold room labeled & covered with aluminum foil overnight10. Wash 6x with working buffer11. Prepare secondary Abs (for confocal immunolabeling)goat anti-rabbit - AF 568 @ 1:50 12. Let sit on cold room shaker covered with foil for 48 hrs13. Wash 3x with 1X PBS 0.814. Prepare nuclear stain* Final solution = equal parts 2xPBS + 1:100 Hoescht 33342 in ddH2O* Prepare each separately. * Once added together, you should not observe any precipitation.* If precipitation is observed.... Do not use the solution!15. 30 min staining with nuclear stain16. Wash 3x with 1X PBS 0.817. Sections mounted on slides and coverslipped using Gelvatol
anterior/posterior region
genetic manipulation
624 days
adult
female
32 grams
mus musculus
mouse
Unspecified
large scale montage at anterior hippocampus
Vibratome
80 m
p1187
Transgenic animal overexpressing alpha-synuclein