Multiscale cardiac imaging spanning the whole heart and its internal cellular architecture in a small animal model
- Biomedical Engineering, Oregon Health & Science University, United States
- Multiscale Microscopy Core, Oregon Health & Science University, United States
- Center for Developmental Health, Knight Cardiovascular Institute, Oregon Health & Science University, United States
Figures
Figure 1
Schematics of steps performed to achieve cardiac multiscale imaging, which yields both 3D whole-heart images and 3D ultrastructural images from the same heart.
Columns depict the four steps employed. In Step 1, the heart is post-fixed with osmium tetroxide to provide contrast for 3D micro-CT images of the whole heart (middle row). Digital sections of the …
Figure 2
Micro-computed tomography (micro-CT) images of a chicken embryo normal heart.
The contrast was achieved by following Step 1 of our correlative multiscale imaging protocol (Figure 1). From left to right: External 3D surface of the heart; cardiac section along the heart’s long …
Figure 3
Comparison of micro-CT images of the two hearts selected for this study.
(A) Segmentations showing the heart morphology for the normal, control (CON) heart and the TOF heart. Red: lumen of the left atrium and ventricle as well as aorta; Blue: lumen of the right atrium …
Figure 4
SBF-SEM images of the control (CON, left) and TOF (right) hearts, indicating the location of imaging.
Overview scans from semithin transverse sections of each heart (top row) were used to confirm uniform stain penetration and the location of 2 ROIs per heart (ROI A and ROI B) further analyzed (red …
Figure 5
Detail of high-resolution SBF-SEM images obtained.
The pictures depict small regions within the selected sub-ROIs from region A of the control (CON) and TOF hearts. Nuclear membranes (N) are intact, as well as myofibrils (Myo) and mitochondria …
Figure 6
Example 3D reconstruction of SBF-SEM image stack acquired from the control heart.
xy is the imaging plane, acquired at 10 nm lateral resolution. z is the depth direction, with xy images acquired every 40 nm. xz and yz are reconstructed perpendicular planes that show the …
Figure 7
Segmented portion of 3D SBF-SEM heart tissue images.
The pictures illustrate the level of detailed ultrastructural architecture that can be obtained from 3D SBF-SEM images. (A and B) show the same 10 × 10 × 10 µm3 of the LV developing chick heart …
Figure 8
Segmentation and quantification of SBF-SEM images.
Selected images (n ≥ 10) from the SBF-SEM image stacks acquired at regions A and B were segmented and quantified. (A) Percentage of myocardial cells occupied by nuclei, myofibrils and mitochondria …
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Figure 8—source data 1
Detailed calculations from segmented SBF-SEM images leading to Figure 8A and B plots.
- https://cdn.elifesciences.org/articles/58138/elife-58138-fig8-data1-v2.xlsx
Figure 9
3D visualization of SBF-SEM segmentations.
(A) Sketch of image dataset acquisition, showing relative orientations. Green plane is the top image, orange plane is the bottom image (last image acquired), blue plane is approximately parallel to …
Figure 10
Myofibril transmural and elliptical angle quantification.
(A) Sketch showing approximate position of the SBF-SEM imaged planes (x-y planes), and image stacks (spanning the z-direction depth) with respect to the heart morphology. For the imaged transmural …
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Figure 10—source data 1
Details of transmural angle θ calculation from SBF-SEM images of the control heart CON A sample.
- https://cdn.elifesciences.org/articles/58138/elife-58138-fig10-data1-v2.xlsx
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Figure 10—source data 2
Details of elliptical angle Φ calculation from SBF-SEM images of the control heart CON A sample.
- https://cdn.elifesciences.org/articles/58138/elife-58138-fig10-data2-v2.xlsx
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Figure 10—source data 3
Details of transmural angle θ calculation from SBF-SEM images of the TOF A heart sample.
- https://cdn.elifesciences.org/articles/58138/elife-58138-fig10-data3-v2.xlsx
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Figure 10—source data 4
Details of elliptical angle Φ calculation from SBF-SEM images of the TOF A heart sample.
- https://cdn.elifesciences.org/articles/58138/elife-58138-fig10-data4-v2.xlsx
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Chemical compound, drug | Sodium Cacodylate | EMS | Cat#RT 12300 | RT 12300 0.1M; pH 7.4 |
| Chemical compound, drug | Tannic acid | EMS | Cat#21700 | 0.1% (w/v) |
| Chemical compound, drug | Osmium Tetroxide, OsO4 | Ted Pella | Cat#18463 | 2% (v/v) final concentration |
| Chemical compound, drug | Potassium Ferricyanide, K₃[Fe(CN)₆] | EMS | Cat#20150 | 1.5% (w/v) |
| Chemical compound, drug | Thiocarbohydrazide, TCH | Aldrich | Cat#223220 | 0.1% (w/v) |
| Chemical compound, drug | Uranyl acetate | Ted Pella | Cat# 19481 | 1% (w/v) working solution |
| Chemical compound, drug | Lead nitrate | EMS | Cat#17900 | Used to prepare Lead Apartate |
| Chemical compound, drug | Aspartic acid | Aldrich | Cat#11195 | Used to prepare Lead Apartate |
| Chemical compound, drug | Acetone | EMS | Cat#10015 | Glass Distilled |
| Chemical compound, drug | Epoxy resin | EMS | Cat# RT 14900 Epon 812 | |
| Software, algorithm | Amira 6.0 | FEI, now ThermoFisher | ||
| Software, algorithm | Dragonfly 4.1 | Object Research Systems |
