Alternate header for print version


Reconstruction
Display image description
Single slice of a processed serial block face SEM imaging data set through the molecular layer of the cerebellum of an adult mouse. Data have been converted from 16 bit to 8 bit and low pass filtered for smoothing.
Full resolution image description
Processed serial block face SEM data set in MRC format. Data have been converted from 16 bit to 8 bit and low pass filtered for smoothing.
File format
MRC
Volume_dimension
6900, 4600, 597
Volume scale
0.0075, 0.0075, 0.06
Animation description
Animation through the slices of a processed serial block face SEM imaging data set through the molecular layer of the cerebellum of an adult mouse. Data have been converted from 16 bit to 8 bit and low pass filtered for smoothing.

Image 2D
Display image description
Single slice from a serial block face imaging data set of the molecular layer of the cerebellum of the adult mouse
Full resolution image description
Tar file containing full resolution raw imaging data in dm3 format
Animation description
Animation through a serial block face imaging data set of the molecular layer of the cerebellum of the adult mouse

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

CCDB:8246*  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
8246
Instrument
FEI Quanta 200 FEG SEM with Gatan 3 View
Microscopy type
Serial Block Face SEM
Product type
SERIAL SECTION
Image basename
cereb8x6
Spatial Axis Image Size Pixel Size
X 6000px 7.5 nm/pixels
Y 4000px 7.5 nm/pixels
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
Recording medium
Gatan backscatter detector
Magification
2500
Accelerating voltage
2.5 kV
Notes
Mag: 2500X Image size: 6000 x 4000 Pixel size (XY): 7.5 nm Step size (Z): 60 nm Beam energy: 2.5 kV Spot size: 2.0 Chamber pressure: high vacuum Pixel dwell time: 16 usec
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.
Imaging product type
Type
Serial section
Cutting plane
parasagittal
Description
Serial block face imaging
Z resolution
60 nm