Alternate header for print version


Reconstruction
Display image description
Single computed slice from a tomographic reconstruction of a 500 nm section through the middle free wall of a cardiac myocyte from the left ventricle of an adult mouse (6-8 months of age), stained with potassium ferrocyanide to increase the contrast of membrane systems.
Full resolution image description
file contains TxBR (projective) generated tomographic volume of heart tissue from mouse left ventricle. Full volume: output.rec, trimmed volume:output.rec_sm, binned down volume: output.rec_bin2. Full resolution volume was used for segmentation.
Downsampled image description
Downsampled version of reconstruction file in IMOD format (~2.0 Gb vs 16 Gb full resolution). Note that segmentation was performed on full resolution volume, available as one of the reconstructions available in the tar file through the Reconstruction download.
File format
MRC
Volume_dimension
2689, 4006, 400
Volume scale
0.00142, 0.00142, 0.00142
Animation description
Animation through the computed slices of a a tomographic reconstruction of a 500 nm section through the middle free wall of a cardiac myocyte from the left ventricle of an adult mouse (6-8 months of age), stained with potassium ferrocyanide to increase the contrast of membrane systems.

Segmentation
Display image description
Manual segmentation of T tubule and SR membranes and mitochondria using IMOD 3.9.3
Segmentation file description
Zip file containing the model file generated by IMOD (WT1_CCDB.mod) and measurements derived from them in IMOD (WT1_CCDB_IMODINFO.txt).

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

CCDB:3603*  Cite 
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
3370
Title
MLP KO
Purpose
purpose
Experimenter(s)
Takeharu Hayashi
Microscopy product
Microscopy product ID
3603
Instrument
JEOL 4000#1
Microscopy type
IVEM
Product type
SINGLE TILT
Image basename
mlp_wt (WT1)
Spatial Axis Image Size Pixel Size
X 2689px
Y 4006px
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
Cell type
myocyte
Imaging parameters
Type
Electron microscopy product
Recording medium
film
Magification
15000
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 gelvatol 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
-58 degrees
Max range
60 degrees
Tilt increment
2 degrees