6541
2009-10-01 00:00:00.0
Eric Bushong
Monica Berlanga
purpose
Ultrastructural analysis of cerebellar and hippocampal neuropil
Animated tilt series of the soma of a protoplasmic astrocyte in a 0.5 um thick section from the hippocampus of a 1 month old male mouse, imaged with intermediate voltage electron microscopy. This section is the 18th in a series of 26 serial semi-thick sections through the astrocyte soma used for a serial tomographic reconstruction (MP7503).
/telescience/home/CCDB_DATA_USER.portal/P2023/Experiment_6395/Subject_6397/Tissue_6399/Microscopy_6541/astrosoma-g3s1_raw.mpg
Electron micrograph of the soma of a protoplasmic astrocyte in a 0.5 um thick section from the hippocampus of a 1 month old male mouse, imaged with intermediate voltage electron microscopy at zero degrees tilt. This section is the 18th in a series of 26 serial semi-thick sections through the astrocyte soma used for a serial tomographic reconstruction (MP7503).
<![CDATA[http://ccdb.ucsd.edu/sand/main?event=rd&type=512r&mpid=6541]]>
File contains all IMOD and TxBR files (st, fid, preali, *com, *log, *mat, etc...) of astrosoma dataset
/telescience/home/CCDB_DATA_USER.portal/P2023/Experiment_6395/Subject_6397/Tissue_6399/Microscopy_6541/astrosoma-g3s1_raw.tar
400 keV
5000
8K CCD
26 section series used for serial tomographic reconstruction
Section 18 of 26
.5 um
54 degrees
-58 degrees
singlet_desc
2 degrees
6541
astrosoma-g3s1
JEOL 4000 #2
IVEM
SINGLE TILT
7072
7072
Sarah Maynard
Eric Bushong
Maryann Martone
Ted Waitt Family Foundation
Mark Ellisman
Stephen Larson
Whole Brain Catalog Project
2009-08-05 00:00:00.0
P2023
Animation through the computed slices of a tomographic reconstruction of a protoplasmic astrocyte in a 0.5 um thick section from the hippocampus of a 1 month old male mouse, imaged with intermediate voltage electron microscopy. This reconstruction is the 18th in a series of 26 serial reconstructions through the cell soma. The complete reconstruction can be viewed under MP7503.
/telescience/home/CCDB_DATA_USER.portal/P2023/Experiment_6395/Subject_6397/Tissue_6399/Microscopy_6541/astrosoma-g3s1_vol.mpg
Single computed slice through a tomographic reconstruction of a protoplasmic astrocyte in a 0.5 um thick section from the hippocampus of a 1 month old male mouse, imaged with intermediate voltage electron microscopy. This reconstruction is the 18th in a series of 26 serial reconstructions through the cell soma. The complete reconstruction can be viewed under MP7503.
<![CDATA[http://ccdb.ucsd.edu/sand/main?event=rd&type=512v&mpid=6541]]>
File contains TxBR generated tomographic reconstructions of astrosoma in MRC format
/telescience/home/CCDB_DATA_USER.portal/P2023/Experiment_6395/Subject_6397/Tissue_6399/Microscopy_6541/astrosoma-g3s1_z_-199.0.out
File contains binned down (by 8 in XYZ) TxBR generated tomographic reconstructions of astrosoma in MRC format. Note the the resolution stated is for the full resolution volume and not the downsampled volume. Binned resolution is 12.24 x 12.24 x 12.24
/telescience/home/CCDB_DATA_USER.portal/P2023/Experiment_6395/Subject_6397/Tissue_6399/Microscopy_6541/astrosoma-g3s1_z_-199.0.out_bin8
MRC
7072,7072,384
.00153,.00153,.00153
protoplasmic astrocyte
brain
soma
2
central nervous system
Perfuse animal with Ringer's solution for 2 minutes followed by 2% glutaraldehyde and 2% paraformaldehyde in 0.15M sodium cacodylate buffer, pH 7.4, at 35 degrees Celsius for 10 minutes. Remove tissue and fix on ice for 1-2 hours. During this time the tissue is further dissected.Wash 5x5 minutes in cold 0.15M sodium cacodylate buffer.Postfix in 1% OsO4 in 0.15M sodium cacodylate buffer on ice for 1 hour.Rinse in cold ddH2O 4x5 minutes.Place tissue into 2% uranyl acetate in ddH2O on ice for 2 hours.Rinse tissue in ice cold ddH2O 3x5 minutes.Dehydrate tissue in ethanol series 20, 50, 70, 90, 100% for 5 minutes each on ice.Complete dehydration with 100% acetone 2x10 minutes, first on ice, second at room temperature.Infiltrate with solution of 50% acetone and 50% durcupan ACM resin overnight in scintillation vials with caps on.Transfer tissue to foil dishes and change to 100% resin for 1 hour. Place into fresh resin for an additional 2 hours.Place in fresh resin and flat embed using 2 mould release treated slides. Polymerize in 60 degree Celsius vacuum oven for 24-48 hours.
Astrocyte Soma Reconstruction
conventional EM
1 months
young adult
male
Harlan
18 grams
Mus musculus
mouse
C57BL/6NHsd
hippocampus, from CA1 stratum radiatum
ultramicrotome
.5 um
embedded
Strong aldehyde and osmium fixation, en bloc uranyl acetate, standard dehydration, and epoxy embedding using a normal male, C57 black 1-month-old mouse.