Building a Multi-Dimensional MAP of Developing Human Lung
Figure 7: IF and 2nd harmonic imaging of lung niches. (A-B) AGER expressing AT1 cells in relationship to SFTPC expressing AT2 cells and a vessel visualized by 2nd harmonics (SHG). (A) 2D image of a single z-stack frame. (B) 3D rendering of the Z-stacks showing the AT1 cell surfaces using Imaris software. (C) 3D representation of the pulmonary vascular network as visualized by cell surface rendering using Imaris software of the capillary bed (PECAM1) and an artery (SHG). (D-E) Visualization of a submucosal gland. ACTA2 stains myofibroblasts. MUC5B stains the mucin made by goblet cells inside the gland surrounded by a collagen matrix detected by (SHG). (D) 2D image of a single Z-stack frame (E) 3D surface rendering in Imaris. See images and videos below.
Figure 7A: AGER expressing Alveolar Type I cells (AT1) cells in relationship to SFTPC expressing AT2 cells and a vessel visualized by 2nd harmonics (SHG).
Panel A: 2D image of a single z-stack frame. (click to enlarge image)
Panel B: 2D video of rendered z-stack frames aquired sequentially. (Click to play video in larger popup window)
A
B
2D video of rendered z-stack frames aquired sequentially showing AT1 AGER expression (green) and AT2 SFTPC (red) expression and collagen from the second harmonics (white) confocal imaging.
Figure 7B: 3D rendering of the Z-stacks.
Panel C: 3D video of rendered z-stacks rendered in Imaris software (Click to play video in larger popup window)
Panel D: 3D rendering of the Z-stacks showing the AT1 cell surfaces using Imaris software. (Click to play video in larger popup window)
C
3D rendering of the Z-stacks showing the AT1 cells expressing AGER (green), the AT2 cells expressing SFTPC (red), and collagen from the second harmonics (white) confocal imaging.
D
3D surface rendering of the Z-stacks showing the AT1 cells expressing AGER (green) and AT2 cells expressing SFTPC (red) and second harmonic (white) confocal imaging.
Figure 7C: 3D representation of the pulmonary vascular network as visualized by cell surface rendering using Imaris software of the capillary bed (PECAM1) and an artery (SHG).
Panel E: 3D video of rendered z-stacks rendered in Imaris software (Click to play video in larger popup window)
Panel F: 3D rendering of the Z-stacks showing the AT1 cell surfaces using Imaris software. (Click to play video in larger popup window)
3D rendering of the Z-stacks showing PECAM1 (red) expression and collagen from the second harmonics (white) confocal imaging.
3D surface rendering of the Z-stacks showing PECAM1 (red) expression and collagen from the second harmonics (white) confocal imaging.
Figure 7D and E: Visualization of a submucosal gland. ACTA2 stains myoepithelial cells. MUC5B stains the mucin made by goblet cells inside the gland surrounded by a collagen matrix detected by (SHG)
Panel G: 2D image of a single Z-stack frames (Click to play video of rendered z stacks in larger popup window)
Panel H: 3D surface rendering in Imaris showing the goblet cells using Imaris software. (Click to play 3D video in larger popup window)
2D rendering of the Z-stacks showing the MUC5B expressing goblet cells (red) , ACTA2 expressing myoepithelial cells (green), and collagen from the second harmonics (white) confocal imaging.
3D surface rendering of the MUC5B expressing goblet cells (red), the ACTA expressing myoepithelial cells (green), and collagen from the second harmonics (white) confocal imaging.
Figure 8: High resolution imaging. (A) TEM of AT2 and surrounding cells. (B) Confocal image showing close proximity of endothelial (SOX17), AT1 (AGER), and mesenchyme (PDGFRb) lung cells in their niches. (C) Confocal image of alveolar wall indicating presence of pericytes and highlighting the close apposition of multiple cell types in alveolar structures. (D-E) High resolution confocal imaging of AT2 cells rendered to show intracellular distribution of SFTPB and ABCA3 in control (D) and a homozygous mutation in ABCA3 that fails to transport lipids (E). Blue staining indicates the nuclei of individual cells. See videos below.
Figure 8D-E: High resolution confocal imaging of AT2 cells rendered to show intracellular distribution of SFTPB and ABCA3 in control and homozygous mutation in ABCA3
Panel D: 3D surface rendering in Imaris of high resolution confocal imaging of AT2 cells rendered to show intracellular distribution of SFTPB and ABCA3 in control human lung. (click to play 3D surface rendered video in popup window)
Panel E: 3D surface rendering in Imaris of high resolution confocal imaging of AT2 cells rendered to show intracellular distribution of SFTPB and ABCA3 in ABCA3 homozygous mutation human lung. (click to play 3D surface rendered video in popup window)
3D surface rendering of high resolution confocal imaging showing cellular localization of SFTPB (green) and ABCA3 (red) in normal human lung.
3D surface rendering of high resolution confocal imaging showing cellular localization of SFTPB (green) and ABCA3 (red) in ABCA3 homozygous mutaion human lung.
