Alternate header for print version


Reconstruction
Display image description
Portion of a single computed slice through a tomographic reconstruction of the myocardium of the left ventricle of a normal mouse. Note that the full reconstruction covers a larger area than is displayed here.
Full resolution image description
Full resolution reconstruction by TxBR (quadratic: WT5aNC1_quad.rec) (~50 Gb). Note that a binned down reconstruction was used for segmentation that is available for download through the Downsampled data file download link.
Downsampled image description
2X binned down version of the reconstruction file in IMOD format. This version was used to generate the segmentation files. Note: to open this file using Jinx, append a .rec to the file name.
File format
MRC
Volume_dimension
4482, 6678, 460
Volume scale
0.0017, 0.0017, 0.0017
Animation description
Animation through the computed slices of a tomographic reconstruction from the left ventricle of a normal mouse heart. The electron dense particles on the surface of the volume are colloidal particles applied to the surface of the imaged section to serve as fiducial markers for alignment of the tilt series.

Segmentation
Display image description
Manual segmentation of T tubule and SR membranes and mitochondria using IMOD 3.9.3
Segmentation file description
file contains IMOD generated tomographic model file (WT5aNC1_CCDB.mod) and information file that contains measurements from the segmentations generated by IMOD (WT5aNC1_CCDB.txt)

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

CCDB:3611*
Project: P1233
Project name
Structural investigation of T-tubule structure in heart failure
Description
3D investigation of T tubule structure using confocal and electron tomography from the MLP knock out mouse and human heart tissue
Leader(s)
Masahiko Hoshijima
Takeharu Hayashi
Collaborator(s)
Andrea Thor
Masako Terada
Maryann Martone
Mark Ellisman
Start date
04-01-2004
End date
unspecified
 
Experiment
Experiment ID
3366
Title
Experiment5
Purpose
purpose
Experimenter(s)
Takeharu Hayashi
Microscopy product
Microscopy product ID
3611
Instrument
JEOL 4000#1
Microscopy type
IVEM
Product type
SINGLE TILT
Image basename
WT5aNC1
Spatial Axis Image Size Pixel Size
X 2242px
Y 3340px
Subject
Species
mouse
Scientific name
MUS MUSCULUS
Strain
S129
Group by
genetic manipulation
Treatment
Control
Age class
adult
Tissue section
Anatomical location
left ventricle
Specimen description
Organ
heart
System
cardiovascular
Structure
T-tubules, sarcoplasmic reticulum, mitochondrion
Cell type
myocyte
Imaging parameters
Type
Electron microscopy product
Recording medium
film
Magification
0
Accelerating voltage
400 kV
Specimen preparation
Protocol used
T-Tubule staining for LM (Confocal Studies)Immersion fix in cold 4% paraformaldehyde in 0.15M sodium cacodylate buffer for 2 hoursRinse tissue in 3x10min1X PBSCut 50um sections on a vibratomeincubate sections in wheat germ agglutinin (WGA) conjugated to FITC (Molcular Probes), diluted 1:100 in phosphate buffered salineincubate sections at 4 C for 1-3 weeks in WGA-FITCrinse sections 3X10min in 1XPBSthen counterstain with propidium iodide (use at 5-10ug/ml concentration) for 30min to reveal cell nucleirinse 3X10min in dH2Ocoverslip in gelvatolEM: T-Tubule or SR Staining of Muscle by Potassium FerrocyanideStandard Fixation: 2% paraformaldehyde and 2% glutaraldehyde in 0.15M sodium cacodylate (Add10mM calcium chloride for SR staining).Block tissue into small pieces.Post Fix: In same fixative for 1-3 hours.Wash 3x10min in 0.15M sodium cacodylate.Incubation in: 0.8% potassium ferrocyanide and 2% osmium tetroxide in 0.15M sodium cacodylate for at least 2 hours (we standardly do overnight) at room temperature.Wash 2 hours-overnight in 0.15M sodium cacodylate.En bloc staining with 1% uranyl acetate 30min.Brief wash in DDH2O.Standard dehydration and embedding.
Imaging product type
Type
Single tilt
Description
singlet_desc
Min range
-60 degrees
Max range
60 degrees
Tilt increment
2 degrees