|
|
Blood, Vol. 111, Issue 3, 1147-1156, February 1, 2008
Live imaging of emerging hematopoietic stem cells and early thymus colonization
Blood Kissa et al.
111: 1147
Supplemental Materials for: Kissa et al
Files in this Data Supplement:
- Video 7. Routes of the first hematopoietic immigrants to the nascent thymus (MOV, 6,832 KB)
-
Time-lapse confocal imaging of GFP (green) and BodipyTR (red) fluorescence; 1 image every 4 min. between 54 hpf and 67 hpf; each image is the projection of a stack of 11 planes, covering a depth of 80 µm. Movie 8 provides the corresponding 3D viewing through a stereogram pair. At t=0, Cell 1 is already visible (blue arrow), next to the optic vein (ov). Cell 2 appears at t=160 min (white arrow), next to the common cardinal vein. At t=420 min., Cell 1 undergoes mitosis (blue arrow), and purple arrows point at the two daughter cells from t=452 min. At the same time point, a yellow arrow points at Cell 3 which came 50 min earlier from just caudal to the ear, and will do the final part of its migration to the thymus together with Cell 2. The trajectories of cell 1 (and its progeny) and cell 2 are recapitulated in the last image.
R, retina; t, thymus; otv, otic vesicle; y, yolk sac; hb, hindbrain; PHS, primary head sinus; CCV, common cardinal vein; ov, optical vein.
- Video 8 (=movie 7-stereo). Routes of the first hematopoietic immigrants to the nascent thymus (MOV, 5,412 KB)
-
To view this stereoscopic version of movie 7, fuse the two images by squinting, wait until stereopsis (3D vision) is attained, then start the movie. This movie is especially useful to see the movements through depth of Cell1 and Cell2.
- Video 9. Extravasation of a CD41-GFPlow precursor from the PHS into the nascent thymus (MOV, 2,839 KB)
-
Spinning disk time-lapse confocal imaging of GFP (green) and BodipyTR (red) fluorescence, in one selected confocal plane at 4 dpf. 1 image every 60 sec. Note that unlike in the other movies presented here, this is the right side (and thymus) of the embryo. Apart from the cell extravasating to the thymus (arrowhead), another GFP+ cell stops in the same zone of the PHS at t=50min59', but is taken away by circulation at t=1h09min.
PHS, primary head sinus; m, thymus-proximal muscle. Scale bar represents 50 µm.
- Video 10. Laser-mediated occlusion of PHS circulation along the nascent thymus (MOV, 782 KB)
-
The first half of the movie shows circulation at 70 hpf in the left PHS of a control embryo; the second half shows the absence of circulation specifically in that vessel in a sibling embryo in which PHS circulation was occluded at 52 hpf by generating a clot upstream, through repeated laser ablation of circulating erythrocytes.
- Video 11. Dynamics of CD41-GFPlow precursors in and out of the nascent thymus at 5 dpf (MOV, 402 KB)
-
Spinning disk time-lapse confocal imaging of GFP (green) and BodipyTR (red) fluorescence. Dots accompany six selected cells with larger (blue dots) or smaller (white dots) amplitude movements. BA, branchial arches (gill circulation); otv, otic vesicle.
|
|
