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Reconstruction
Display image description
maximum intensity projection of tomographic reconstruction of PSDs in area CA1 (blue arrow).Intensity is reversed so that PSDs appear bright against a dark background.
Full resolution image description
Volume reconstruction of selectively stained PSDs in Analyze format
Volume_dimension
451, 411, 238
Volume scale
0.004, 0.004, 0.004
Animation description
rotation loop of maximum intensity projection of shamca1 in 5 degrees. increments along the y axis. The contrast is reversed so that PSD appear bright against a dark background.

Image 2D
Full resolution image description
This tar file contains the original tile images; shamca1.???.f along with the fiducial mark file (shamca1.fido) used to align the data

Segmentation
Display image description
Obj file contains individual PSDs segmented from shamca1.img using the "multiple objects" feature of Analyze.

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

CCDB:21*
Project: P1110
Project name
Synaptic alterations in transient ischemia
Description
Investigation of morphological alterations of post-synaptic densities following an episode of transient ischemia
Funding agency
NIH
Leader(s)
Bingren Hu
Collaborator(s)
Maryann Martone; Ying Jones
Start date
01-01-1995
End date
unspecified
 
Experiment
Experiment ID
13
Experiment date
05-01-1997
Title
Tomographic reconstruction of EPTA stained post-synaptic densities
Purpose
3D structure of post-synaptic densities selectively stained with EPTA
Experimenter(s)
Bingren Hu
Microscopy product
Microscopy product ID
21
Instrument
JEOL4000 IVEM
Microscopy type
IVEM
Product type
single tilt
Image basename
shamca1
Spatial Axis Image Size Pixel Size
X 1024px 0.004 µm
Y 1024px 0.004 µm
Subject
Species
rat
Scientific name
rattus norvegicus
Strain
Wistar
Group by
time of reperfusion
Age class
adult
Tissue section
Anatomical location
hippocampus
Microtome
ultramicrotome
Thickness
1 µm
Specimen description
Organ
brain
System
central nervous system
Structure
post synaptic density
Imaging parameters
Type
Electron microscopy product
Recording medium
film
Magification
20000
Accelerating voltage
400 KeV
Specimen preparation
Protocol used
Ischemia model: All experimental procedures were approved by theSubcommittee on Animal Studies of the Veterans Affairs Medical Center(San Diego, CA). Male Wistar rats (250-300 gm) were fasted overnight.Anesthesia was induced with 3% halothane followed by maintenance with1-2% halothane in an oxygen/nitrous oxide (30/70%) gas mixture.Catheters were inserted into the external jugular vein, tail artery,and tail vein to allow blood sampling, arterial blood pressurerecording, and drug infusion. Both common carotid arteries wereexposed and encircled by loose ligatures. Fifteen minutes beforeischemia induction and 15 min after ischemia, blood gases weremeasured and adjusted to PaO2 >90 mmHg and PaCO2 35-45 mmHg, pH7.35-7.45, by adjusting tidal volume of the respirator. Bipolar EEGwas recorded every 5-10 min before ischemia, continuously during theischemic insult, and every 5 min after ischemia until the ratrecovered from the anesthesia. At the beginning of a 30 minsteady-state period before induction of ischemia, the inspiredhalothane concentration was decreased to 0.5%, and 150 IU/kg heparinwas administered intravenously. Blood was withdrawn via the jugularcatheter to produce a mean arterial blood pressure of 50 mmHg, andischemia was induced by clamping both carotid arteries. Bloodpressure was maintained at 50 mmHg during the ischemic period by2end of the ischemic period, the clamps were removed, and the bloodwas reinfused through the jugular catheter, followed by 0.5 ml of 0.6M sodium bicarbonate. In all experiments, temperature was maintainedat 37C before, during, and after ischemia (15 min of reperfusion).Halothane was discontinued at the end of ischemia, and all woundswere sutured. At 4 or 24 hr, 3 d, or 1 week after the ischemicepisode, the animals were reanesthetized, tracheotomized, andartificially ventilated. For electron microscopic studies, the brainswere perfused with ice-cold 2% paraformaldehyde and 2.5%glutaraldehyde in 0.1 M cacodylate buffer. Sham-operated control ratswere subjected to the same surgical procedures but without inductionof ischemia.Electron microscopic studies: Tissue sections fromexperimental and control animals were stained either by 1% ethanolicphosphotungstic acid (E-PTA) (Bloom and Aghajanian, 1966, 1968) or bythe conventional osmium-uranium-lead method. Briefly, coronal brainsections were cut at a thickness of 200 um with a Vibratome throughthe level of the dorsal hippocampus and post-fixed for 1 hr with 4%glutaraldehyde in 0.1 M cacodylate buffer, pH 7.4. For conventionalosmium-uranium-lead staining, sections were post-fixed for 2 hr in 1%osmium tetroxide in 0.1 M cacodylate buffer, rinsed in distilledwater, and stained with 1% aqueous uranyl acetate overnight. Thetissue sections were then dehydrated in an ascending series ofethanol to 100%, followed by dry acetone, and embedded in DurupanACM. Thin sections were counterstained with lead citrate beforeexamination in the electron microscope. For E-PTA staining, sectionswere dehydrated in an ascending series of ethanol to 100% and stainedfor 1 hr with 1% PTA prepared by dissolving 0.1 gm of PTA in 10 ml of100% ethanol and adding four drops of 95% ethanol. Sections were thenembedded in Durcupan ACM.
Imaging product type
Type
Single tilt
Description
singlet_desc
Min range
-60 degrees
Max range
60 degrees
Tilt increment
2 degrees


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