FIGURE SUMMARY
Title

In-vivo 3D imaging of Zebrafish's intersegmental vessel development by a bi-directional light-sheet illumination microscope

Authors
Qin, X., Chen, C., Wang, L., Chen, X., Liang, Y., Jin, X., Pan, W., Liu, Z., Li, H., Yang, G.
Source
Full text @ Biochem. Biophys. Res. Commun.

Schematic diagram of bi-directional light-sheet illumination microscope. (A) L1-L4, lens. M1-M4: Mirrors; CO: collimation objective; BS: beam split; SL1-SL2: scanning lens; Gavlo1-Gavol2: Gavlo mirror pairs; IO1-IO2: illumination objectives; DO: detection objective; TL: tube-lens. (B)–(C) The spreading of the light sheet caused by sample scattering. The blue arrows indicate the illuminating direction. (D) The weighted fusion of the two images from left- and right-illuminated light sheets. For the image generated by illuminating from the left side, only the left half of the outline (red solid line) remains, and the other half is replaced by the right-side outline (green solid line) generated by right illumination. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

3D light-sheet imaging of vascular structure in a Zebrafish trunk from 38–60 hpf with a time interval of 2 h. Insets show the corresponding 2D MIP images. Scale bar: 100 μm.

Vessel Length measurements by MIP- and VR-based methods. (A) 3D reconstruction of the vasculature in the developing Zebrafish trunk at approximately 62 hpf. The orange arrow represents the projection direction for MIP. (B) The 2D MIP image of trunk vessels at projection angle 0° shows both ISVs and DLAVs where two DLAVs overlap. Scale bar: 100 μm. (C) The measured length of the 3rd and 7th pairs of ISVs from 2D MIP images dependent on projection direction. (D) 3D reconstruction of the 6th and 7th pairs of ISVs shown in (A) and (B). (E) The MIP image of the 6th and 7th pairs of ISVs in the Y direction. Scale bar: 50 μm. (F) The measured lengths of the 2nd to 7th left ISVs with MIP and VR. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

The growth curve of the left DLAV and ISVs in a developing Zebrafish embryo at 38–62 hpf. (A) The growth curve of the left DLAV respectively based on MIP and VR. (B) The growth curve of 2nd to 7th pairs of ISVs based on VR. (C–H) The 3D images of 2nd to 4th pair of ISVs from 38 hpf to 48 hpf.

Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Biochem. Biophys. Res. Commun.