Alternate header for print version


Reconstruction
Display image description
Single slice from a cropped and contrast adjusted serial section reconstruction from the molecular layer of the cerebellar vermis of the adult mouse generated from serial block face SEM.
Full resolution image description
Cropped and contrast adjusted serial block face imaging volume in MRC format.
File format
MRC
Volume scale
0.0056, 0.0056, 0.05
Animation description
Downsampled animation through the cropped and contrast adjusted serial section reconstruction from the molecular layer of the cerebellar vermis of the adult mouse generated from serial block face SEM.

Image 2D
Display image description
Single slice from a serial section reconstruction of the cerebellar molecular layer neuropil in the vermis of the adult mouse cerebellum, generated from serial block face SEM imaging.
Full resolution image description
Serial block face raw imaging data set in mrc format. A cropped and contrast adjusted volume is available through the reconstruction form.

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

CCDB:8192*  Cite 
Project: P2080
Project name
3View volumes
Description
Various volumes generated on the 3View during testing or as preliminary data for other projects
Leader(s)
Eric Bushong
Collaborator(s)
Tom Deerinck
Start date
unspecified
End date
unspecified
 
Experiment
Experiment ID
8186
Title
cerebellum
Purpose
purpose
Experimenter(s)
Eric Bushong
Microscopy product
Microscopy product ID
8192
Instrument
Gatan 3View system on FEI Quanta microscope
Microscopy type
Serial Block Face SEM
Product type
SERIAL SECTION
Image basename
080309
Spatial Axis Image Size Pixel Size
X 4090px 5.6 nm/pixels
Y 4096px 5.6 nm/pixels
Y 401px
Subject
Species
Mouse
Scientific name
mus
Strain
C57BL/6
Group by
Age
Age
2 months
Age class
adult
Tissue section
Anatomical location
Cerebellum
Specimen description
Organ
brain
System
central nervous system
Structure
neuropil
Imaging parameters
Type
Electron microscopy product
Magification
5000
Accelerating voltage
2 kV
Notes
image size: 4090 x 4096 x ?? (X x Y x Z) pixel size: 5.6 nm XY, 50 nm Z magnification: 5000X beam energy: 2 kV spot size: 2.5 dwell time: 10 usec chamber pressure: 10 Pa
Specimen preparation
Protocol used
The mouse was anesthetized with Nembutal and then perfused with Ringer's containing heparin and xylocaine and bubbled with 95% O2 / 5% CO2. The mouse was then perfused with 2.5% glutaraldehyde / 2% PFA in cacodylate buffer containing 2 mM CaCl.The brain was removed and placed in same post-fix for 2 hours and 15 minutes in fridge.The brain was cut in to 100 um thick vibratome sections in ice cold cac buffer with 2mM Ca.The slices were washed in cac w/ Ca for 30 minutes in fridge.The slices were placed into 2% OsO4 / 1.5% potassium ferrocyanide in cac w/ Ca at room temp for 1 hour.The slices were washed 3x 2 minutes with ddH2O and then placed into 1% aq. thiocarbohydrazide for 20 minutes at room temp.The slices were washed 2x 2 minutes with ddH2O and then placed in 2% aq. OsO4 for 30 min at room temp.The slices were washed 3x 2 min with ddH2O and then placed in 2% aq. UA overnight in fridge.The next day the slices were washed 3x 10 min in ddH2O and then placed in lead aspartate solution in 60 degree oven for 30 min.The slices were washed 3x 5min in ddH2O at room temp.The slices were dehydrated in series of ice cold EtOH solutions (70%, 90%, 100%, 100%; 10 min each).The slices were placed in ice-cold acetone for 10 min and left at room tempThe slices were placed in room temp acetone for 10 minutes.The slices were placed in 25% Durcupan for 3 hours. The slices were placed in 50% Durcupan overnight.The slices were placed in 75% Durcupan for 3 hours. The slices were placed in 100% Durcupan overnight.The slices were flat-embedded in fresh 100% Durcupan and left in 50 degree oven for 48 hours.